Zhuo et al looked into the part of transmembrane 9 superfamily member 1(TM9SF1)in bladder cancer(BC),and evaluated if it can be used as a therapeutic target.They created a permanent BC cell line and tested the effects...Zhuo et al looked into the part of transmembrane 9 superfamily member 1(TM9SF1)in bladder cancer(BC),and evaluated if it can be used as a therapeutic target.They created a permanent BC cell line and tested the effects of TM9SF1 overexpression and suppression on BC cell growth,movement,invasion,and cell cycle advancement.Their results show that TM9SF1 can boost the growth,movement,and invasion of BC cells and their access into the G2/M stage of the cell cycle.This research gives a novel direction and concept for targeted therapy of BC.展开更多
Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon ...Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon cancer cell line CCL229. Methods: Using the molecular and biochemical techniques and electron microscopy to analyze the expression of iNOS, production of NO and growth characteristics of human colon cancer cells. Results: cytokine treatment can induce expression of the iNOS gene and production of nitric oxide was significantly higher after treatment of CCL229 cells with TNF-αor IFN-γ. Treatment with either cytokine or a combination of both significantly increased levels of Malondialdehyde (MDA) over control. Furthermore, cytokine treatment increased the proliferation inhibition rate as assessed in vitro and decreased the cell proliferation index on flow cytometry. Electron microscopy showed that cells treated with cytokines had fewer pseudopodia or cell processes than control cells and that cytokine treated cells had dilatation of the mitochondria and endoplasmic reticulum and dilated vesicular or tubular cisternae. Conclusion: Our findings indicate that TNF-α and IFN-γ induce the expression of iNOS gene in CCL229 cells, which increases the production of nitric oxide, inhibits proliferation, causes lipid peroxidation, and results in ultrastructural changes.展开更多
The undecapeptide substance P(SP) was shown to be intimately involved in both the structural and functional aspects of the anterior pituitary. Yet, in addition to its influences on hormonal secretion, SP may well poss...The undecapeptide substance P(SP) was shown to be intimately involved in both the structural and functional aspects of the anterior pituitary. Yet, in addition to its influences on hormonal secretion, SP may well possess more actions in this master gland. The present study was ftherefore aimed to investigate the effect of SP on the proliferation of rat anterior pituitary cells in primary culture. It was found that SP could dose-dependently increase the ineorporation of tritiated thymidine (3H-TdR)into cultured anterior pituitary cells. Other mammalian tachykinins such as neurokinin A and neurokinin B had similar effect but to varying degrees. The equipotent analogue of SP, Norleucine(11) -SP(Nle(11)-SP), also acted likewise, with its action antagonizable by spantide, a SP receptor blocker. To further characterize the nature of cells responsive to the challenge of SP, immunocytochemical staining against S-100 protein and some adenohypophyseal hormones was performed alone or plus autoradiography The results showed that the percentage of S-100 proteinimmunoreactive cells was apparently elevated by the addtion of Nle(11)-SP for 48 h, which indicates a preferential proliferation of folliculo-stellate cells under the regime. This was confirmed by increases in immunocytochemical or autoradiographical labelling indices of anterior pituitary cells treated similarly. Taken togethbr, these results reveal that the trophic action of SP observed previously in other tissues is also present at least in cultured rat anterior pituitary cells, with responding cells being predominantly folliculo-stellate cells as typified by S-100 proteinimmunoreactivity. Therefore, an intra-pituitary trophic action of SP in vivo could be anticipated.展开更多
Astrocytes play a major role in the reactive processes in response to neuronal injuries in the brain. Excessive gliosis is detrimental and can contribute to neuronal damage. CDS1 (TAPA), a member of the tetraspanin ...Astrocytes play a major role in the reactive processes in response to neuronal injuries in the brain. Excessive gliosis is detrimental and can contribute to neuronal damage. CDS1 (TAPA), a member of the tetraspanin family of proteins, is upregulated by astrocytes after traumatic injury to the rat central nervous system (CNS). To further understand the role of CD81 in the inhibition of astrocytes, we analyzed the effects of a CD81 antibody, on cultured rat astrocytes. The results indicated that the effect worked in a dose-dependent manner with certain dosage range. It, however, reached a dosage equilibrium at a high dosage. Furthermore, anti-CD81 antibody remarkably inhibited the proliferation of astrocytes after incubation with astrocytes for different periods of time and the effect presented a time-dependent fashion. However, anti-CDS1 antibody substantially inhibited the proliferation of astrocytes at low density and middle density but slightly inhibited the proliferation of as- trocytes at high density, suggesting that the effect was positively correlated with the proliferative ability of astrocytes. Finally, the cell cycle of astrocytes exposured to anti-CD81 antibody was arrested in S phase at the initial stage and at G0/GI phase over time. These findings indicated that CD81 exert significant inhibitory effect, dose-dependently and time-dependently, on the proliferation of astrocytes and the effect is positively correlated with the proliferative capability of astrocytes.展开更多
Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequenc...Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequencing,and then cloned into the eukaryotic expression vector pIRES2-EGFP,which encodes green fluorescent protein(GFP),to construct p1RES2-EGFP-NDRG2 plasmid.OS-RC-2 cells with NDRG2 negative expression were transfected with p1RES2-EGFP-NDRG2 plasmid.The growth of transfected OS-RC-2 cells was observed under light and fluorescence microscopes.After colony-forming cell assays,cell proliferation detection and MTT assays,the growth curves of cells in each group were plotted to investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of OS-RC-2 cells.Cell cycle was determined by flow cytometry.Confocal laser scanning microscopy showed that NDRG2 protein was specifically located on subcellular organelle.Results:A eukaryotic expression vector p1RES2-EGFP-NDRG2 was successfully constructed.After NDRG2 transfection,the growth of OS-RC-2 cells was inhibited.Flow cytometry showed that cells were arrested in S phase but the peak of cell apoptosis was not present,and confocal laser scanning microscopy showed that NDRG2 protein was located in mitochondrion.Conclusions:NDRG2 can significantly inhibit the proliferation of OS-RC-2 cells in vitro and its protein is specifically expressed in the mitochondrion.展开更多
The effects of bleomycin A5 (BLM A5) alone and combined with calmodulin inhibitor N-(4-aminobutyl)-5-chloro-2-naphthalene sulfonamide (W-13) on the proliferation on S-180 cells in vitro were studied. IC50 of BLM used ...The effects of bleomycin A5 (BLM A5) alone and combined with calmodulin inhibitor N-(4-aminobutyl)-5-chloro-2-naphthalene sulfonamide (W-13) on the proliferation on S-180 cells in vitro were studied. IC50 of BLM used alone for the cells was about 2.63 μg/ml, but it was reduced to 1/3.8 and 1/9.5 of 2.63 μg/ml when plus W-13 1, 5 μg/ml respectively. The results indicated that nontoxic doses of W-13 enhanced the hinibition of cell proliferation under the condition of BLM 0.5 - 2.5 μg/ ml. In colony forming test, the survival fraction of S-180 cells treated with BLM plus W-13 was decreased to 1/87 - 240 of that of the cells treated with BLM alone. The results suggest that W-13 can enhance antitumor activity of BLM in vitro and may be used as an synergist of BLM A5 in vivo.展开更多
BACKGROUND Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages.In humans,their limited numbers pose a challenge in fulfilling the...BACKGROUND Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages.In humans,their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs.Studies suggested that mesenchymal stem cells(MSCs),necessary for repair and regeneration via transplantation,require doses ranging from 10 to 400 million cells.Furthermore,the limited expansion of MSCs restricts their therapeutic application.AIM To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.METHODS Human umbilical cord(hUC)tissue derived MSCs were obtained and re-cultured.These cultured cells were subjected to the following evaluation pro-cedures:Immunophenotyping,immunocytochemical staining,trilineage differentiation,population doubling time and number,gene expression markers for proliferation,cell cycle progression,senescence-associatedβ-galactosidase assay,human telomerase reverse transcriptase(hTERT)expression,mycoplasma,cytomegalovirus and endotoxin detection.RESULTS Analysis of pluripotent gene markers Oct4,Sox2,and Nanog in recultured hUC-MSC revealed no significant differences.The immunophenotypic markers CD90,CD73,CD105,CD44,vimentin,CD29,Stro-1,and Lin28 were positively expressed by these recultured expanded MSCs,and were found negative for CD34,CD11b,CD19,CD45,and HLA-DR.The recultured hUC-MSC population continued to expand through passage 15.Proliferative gene expression of Pax6,BMP2,and TGFb1 showed no significant variation between recultured hUC-MSC groups.Nevertheless,a significant increase(P<0.001)in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs.Cellular senescence markers(hTERT expression andβ-galactosidase activity)did not show any negative effect on recultured hUC-MSCs.Additionally,quality control assessments consistently confirmed the absence of mycoplasma,cytomegalovirus,and endotoxin contamination.CONCLUSION This study proposes the development of a novel protocol for efficiently expanding stem cell population.This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies.展开更多
A study was done to produce a rapid in vitro propagation of three strawberry genotypes and tested in the field under Bangladeshi circumstances. Festival, RABI-3, and Neho strawberry genotypes’ runner tips were cultiv...A study was done to produce a rapid in vitro propagation of three strawberry genotypes and tested in the field under Bangladeshi circumstances. Festival, RABI-3, and Neho strawberry genotypes’ runner tips were cultivated in vitro to induce root induction and multiple shoot proliferation. MS (Murashige and Skoog) media that were basally containing three different concentrations at 1.0, 1.5, and 2.0 of BA (6-benzyl adenine), KIN (6-furfuryl amino purine), or GA<sub>3</sub> (gibberellic acid) at 0.5 mg/L increasing tips of the runner was attained. The culture grew on the medium provided with 1.5 mg/L 6-benzyl adenine and 0.5 mg/L 6-furfuryl amino acid to increase shoot at the best level. Micro-cuttings were rooted on MS media at half strength combined with 0.5 mg/L - 1.5 mg/L IBA (indole butyric acid) or IAA (indole acetic acid). IBA attained 4 - 9 roots and 91% - 96% rooting at 1.0 mg/L. The resulting plantlets grew into hardy plants and took root in the earth. The genotype festival had the highest response rate, followed by RABI-3 and Neho.展开更多
BACKGROUND Zinc(Zn)is the second most abundant trace element after Fe,present in the human body.It is frequently reported in association with cell growth and proliferation,and its deficiency is considered to be a majo...BACKGROUND Zinc(Zn)is the second most abundant trace element after Fe,present in the human body.It is frequently reported in association with cell growth and proliferation,and its deficiency is considered to be a major disease contributing factor.AIM To determine the effect of Zn on in vitro growth and proliferation of human umbilical cord(hUC)-derived mesenchymal stem cells(MSCs).METHODS hUC-MSCs were isolated from human umbilical cord tissue and characterized based on immunocytochemistry,immunophenotyping,and tri-lineage differentiation.The impact of Zn on cytotoxicity and proliferation was determined by MTT and Alamar blue assay.To determine the effect of Zn on population doubling time(PDT),hUC-MSCs were cultured in media with and without Zn for several passages.An in vitro scratch assay was performed to analyze the effect of Zn on the wound healing and migration capability of hUC-MSCs.A cell adhesion assay was used to test the surface adhesiveness of hUC-MSCs.Transcriptional analysis of genes involved in the cell cycle,proliferation,migration,and selfrenewal of hUC-MSCs was performed by quantitative real-time polymerase chain reaction.The protein expression of Lin28,a pluripotency marker,was analyzed by immunocytochemistry.RESULTS Zn at lower concentrations enhanced the rate of proliferation but at higher concentrations(>100μM),showed concentration dependent cytotoxicity in hUC-MSCs.hUC-MSCs treated with Zn exhibited a significantly greater healing and migration rate compared to untreated cells.Zn also increased the cell adhesion rate,and colony forming efficiency(CFE).In addition,Zn upregulated the expression of genes involved in the cell cycle(CDC20,CDK1,CCNA2,CDCA2),proliferation(transforming growth factorβ1,GDF5,hypoxia-inducible factor 1α),migration(CXCR4,VCAM1,VEGF-A),and self-renewal(OCT4,SOX2,NANOG)of hUC-MSCs.Expression of Lin28 protein was significantly increased in cells treated with Zn.CONCLUSION Our findings suggest that zinc enhances the proliferation rate of hUC-MSCs decreasing the PDT,and maintaining the CFE.Zn also enhances the cell adhesion,migration,and self-renewal of hUC-MSCs.These results highlight the essential role of Zn in cell growth and development.展开更多
AIM: To explore whether resveratrol (Res) can inhibit human retinal pigment epithelial cell (ARPE-19 cell) proliferation and migration, and to research the molecular mechanisms.METHODS: ARPE-19 cells were pretre...AIM: To explore whether resveratrol (Res) can inhibit human retinal pigment epithelial cell (ARPE-19 cell) proliferation and migration, and to research the molecular mechanisms.METHODS: ARPE-19 cells were pretreated with various concentrations at 0, 50, 100, 150, 200 and 300 μmol/L of Res, and with 0 μmol/L Res as the control for 24, 48 and 72h. The cell proliferation, apoptosis and migration were measured with cell counting kit-8 (CCK-8), flow cytometry, and wound-healing and Transwell assays, respectively. The expression of proliferating cell nuclear antigen (PCNA), P21 and P27, as well as matrix metalloproteinase-9 (MMP-9) and p38 mitogen-activated protein kinases (p38MAPK) was identified by Western blot.RESULTS: Cell proliferation was effectively inhibited by Res (P〈0.05). When pretreated with Res, cells arrested in S-phase increased remarkably (P〈0.05), but the apoptosis ratios showed no significant difference between the treatment and control groups (P〉0.05). Cell migration was suppressed by Res both in wound-healing assay and Transwell migration assay (P〈0.05). Decreases of PCNA, MMP-9 and p38MAPK, as well as increases of P21 and P27 were detected by Western blot (P〈0.05). CONCLUSION: Res can inhibit APRE-19 cell proliferation and migration in a concentration-dependent manner with up-regulation of the expression of P21 and P27, and down-regulation of PCNA, MMP-9 and p38MAPK.展开更多
AIM: To evaluate the effects of adenovirus-mediated gene transfer of RhoA siRNA and RhoC siRNA on proliferation and invasion of SGC7901 cells by Rho/ PI3K/Akt pathway. METHODS: Plasmid of RhoA siRNA and RhoC siRNA wer...AIM: To evaluate the effects of adenovirus-mediated gene transfer of RhoA siRNA and RhoC siRNA on proliferation and invasion of SGC7901 cells by Rho/ PI3K/Akt pathway. METHODS: Plasmid of RhoA siRNA and RhoC siRNA were constructed and transfected into SGC7901 cells. siRNA and LY294002 (PI3K inhibitor) were designed as the control group. The mRNA and protein expressions of RhoA and RhoC were respectively detected with RT-PCR and western blotting. In order to fi nd out the changes of proliferation and invasion power of SGC7901 cell lines, we analyzed the data by MTT, Boyden chamber and evaluated apoptosis of cell with flow cytometry. We treated BALB /C nude mice with RhoA and RhoC-siRNA, and tumor control rate (%) in nude mice was calculated. RESULTS: RhoA and RhoC siRNA transfections specifically down-regulated the corresponding mRNA and protein levels in SGC7901 Cells.The experiment of permeated artificial basal membrane showed that the invasion power of SGC7901 cell lines are on the decline after treatment of Ad-RhoA and RhoC-siRNA (12.64 ± 3.27 vs 87.38 ± 17.38, P < 0.05). The values of 490 nm wavelength light absorption were different in the five groups. The number of alive cells in the group of RhoA and RhoC-siRNA was lower than others in the 6th d (0.71 ± 0.01 vs 3.82 ± 0,11 P < 0.05). The apoptosis rate of transfected RhoA and RhoC-siRNA group with FACS were 19.07% ± 1.78 and there were significant differences between treated and control groups (19.07 ± 1.78% vs 1.23 ± 0.11%, P < 0.01). The tumor transplantation experiment in BALB/C nude mice showed intratumoralinjection of RhoA or RhoC siRNA can inhibit tumor growth. CONCLUSION: RhoA and RhoC siRNA gene therapy mediated by adenovirus may be useful for inhibiting growth and invasion of SGC7901 through a PI3K/Akt pathway. These results provide a novel therapeutic target in preventing gastric cancer cell invasion and metastasis.展开更多
The effects of DK2,a peroxisome proliferator-activated receptor γ agonist,on cultured human pterygium fibroblasts (HPFs) in virto were studied.The HPFs were incubated with 0-200 μmol/L DK2 for 12-72 h.The MTT method...The effects of DK2,a peroxisome proliferator-activated receptor γ agonist,on cultured human pterygium fibroblasts (HPFs) in virto were studied.The HPFs were incubated with 0-200 μmol/L DK2 for 12-72 h.The MTT method was used to assay the bio-activity of DK2 at different doses and time.The cytotoxic effect of DK2 was measured by LDH release assay.The cell cycle distribution and apoptosis were flow cytometrically detected.The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by real-time PCR (RT-PCR) and Western blotting.The results showed that administration of 1-75 μmol/L DK2 for 12-72 h could significantly inhibit HPF proliferation in a dose-and time-dependent manner.DK2-treated cells did not release significant amount of LDH as compared with rosiglitazone-treated cells.After treatment with DK2 at concentrations of 15,25 μmol/L for 24 h,the number of HPFs in G 0 /G 1 phase was significantly increased while that in S phase was significantly decreased (P【0.05),leading to arrest at G 0 /G 1 phase.The apoptosis rates of HPF cells in drug-treated groups were significantly higher than the rate of control group (P【0.05).At the dosage range between 15-25 μmol/L,DK2 could inhibit the expression of PCNA mRNA and protein in HPFs in a dose-dependent fashion (P【0.05).It was concluded that PPARγ agonist can significantly inhibit HPF proliferation,resulting in the arrest at G 0 /G 1 phase,induce the apoptosis of HPFs,and suppress the synthesis of PCNA,in dose-and time-dependent manners.展开更多
Neural stem cells promote neuronal regeneration and repair of brain tissue after injury,but have limited resources and proliferative ability in vivo.We hypothesized that nerve growth factor would promote in vitro prol...Neural stem cells promote neuronal regeneration and repair of brain tissue after injury,but have limited resources and proliferative ability in vivo.We hypothesized that nerve growth factor would promote in vitro proliferation of neural stem cells derived from the tree shrews,a primate-like mammal that has been proposed as an alternative to primates in biomedical translational research.We cultured neural stem cells from the hippocampus of tree shrews at embryonic day 38,and added nerve growth factor(100 μg/L) to the culture medium.Neural stem cells from the hippocampus of tree shrews cultured without nerve growth factor were used as controls.After 3 days,fluorescence microscopy after DAPI and nestin staining revealed that the number of neurospheres and DAPI/nestin-positive cells was markedly greater in the nerve growth factor-treated cells than in control cells.These findings demonstrate that nerve growth factor promotes the proliferation of neural stem cells derived from tree shrews.展开更多
The effect of 1,25-dihydroxyvitamin D<sub>3</sub> [1,25(OH)<sub>2</sub>D<sub>3</sub>] and13-cis-retinoic acid(RA)on the proliferation of a novel human megakaryoblasticleukemia c...The effect of 1,25-dihydroxyvitamin D<sub>3</sub> [1,25(OH)<sub>2</sub>D<sub>3</sub>] and13-cis-retinoic acid(RA)on the proliferation of a novel human megakaryoblasticleukemia cell line(HIMeg)was investigated.At the concentration of 10<sup>9</sup> 10<sup>6</sup>mol/L,1,25(OH)<sub>2</sub>D<sub>3</sub> and RA showed significant inhibition of the proliferation of themegakaryoblastic leukemic cells,which was demonstrated by the count of survival cells,incorporation of<sup>3</sup>H-TdR and<sup>3</sup>H-UR,and cloning efficiency in dose-dependent and time-dependent manners.The results can further explain the mechanism of differentiation-inducing agents and the effect of 1 ,25(OH)<sub>2</sub>D<sub>3</sub> on myelofibrosis.It is possible for1,25(OH)<sub>2</sub>D<sub>3</sub> and RA to be used to treat malignant megakaryocytic diseases.展开更多
Summary: The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx Hep-2 in vitro and their corresponding mechanisms were investigated. Hep-2 cells ...Summary: The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx Hep-2 in vitro and their corresponding mechanisms were investigated. Hep-2 cells were cultured with two kinds of selective cyclooxygenase-2 inhibitors (Sc-58125 and Celecoxib) at various concentrations for 24 h. Morphological changes were observed under the phase microscopy and the growth suppression was detected by using MTT colorimetric assay. Apoptotic DNA fragments were observed by agarose gel electrophoresis, and the cell cycle and apoptotic rate were detected by flow cytometry (FCM) respectively. Hep-2 cells became rounded and detached from the culture dish after being treated with Celecoxib for 24 h, however, they remained morphologically unchanged with Sc-58125. Sc-58125 could increase G 2 phase cells, whereas, Celecoxib rose G 1 phase cells. Both of the two effects were dose-dependent. Moreover, the Hep-2 cells cultured with 50 μmol/L and 100 μmol/L Celecoxib showed obvious apoptosis, with the nuclear DNA of cells exhibiting characteristic DNA ladder. So Sc-58125 could inhibit the proliferation of Hep-2 cells by altering the G 2 phase cells. However, Celecoxib had the same effect by changing the G 1 phase cells and inducing apoptosis at higher concentration.展开更多
Acacia karroo Hayne is an arbor species widely distributed in South Africa with the characteristics of fast growth and drought resistance. The species was introduced to China recently. In vitro culture is an effective...Acacia karroo Hayne is an arbor species widely distributed in South Africa with the characteristics of fast growth and drought resistance. The species was introduced to China recently. In vitro culture is an effective method to rapidly produce plants and a strategy to minimize somaclonal variation among regenerated plants. Browning, however, is a problem in establishing the in vitro culture system. The present study diminished the problem by selecting explants, using different browning inhibitors and chilling treatment. Results showed that the use of embryos as explants reduced the browning ratio to 46.7%, whilst stem segment explants were browned up to 56.7%. The adventitious buds were successfully induced in the modified tissue culture medium supplemented with 5.0 mg·L^-1 6-BA and 0.1 mg·L^-1 NAA. The proliferation coefficient of adventitious buds is 2.8.展开更多
Objective:To observe the effects of platelet-rich plasma(PRP)on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue.Methods:Fibroblasts were separated from human ...Objective:To observe the effects of platelet-rich plasma(PRP)on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue.Methods:Fibroblasts were separated from human chronic refractory wound granulation tissue and then were identified.The obtained fibroblasts were divided into fetal bovine serum(FBS)group,hydrogel group and PRP group,and the three groups were cultured with culture mediums containing FBS,hydrogel and PRP respectively,in order to observe the growth of fibroblasts.The wound scratch assay was used to observe the migration of fibroblasts.Results:PRP group had more fibroblasts than FBS group and hydrogel group since Day 5 of culture,and exhibited greater fibroblast scratch migration area than FBS group on 48 h and 72 h of wound scratch assay(all p<.05).Conclusions:Compared with FBS,human fibroblasts cultured by PRP can more effectively promote the proliferation and migration of fibroblasts.展开更多
The inhibition of a-glucosidase and glucose-6-phosphatase,two enzymes involved in the carbohydrate metabolism,is an important target to control glycaemia on individuals with type 2 diabetes.In this work we report for ...The inhibition of a-glucosidase and glucose-6-phosphatase,two enzymes involved in the carbohydrate metabolism,is an important target to control glycaemia on individuals with type 2 diabetes.In this work we report for the first time the inhibition of both enzymes by the antihyperglycemic n-butanol extract from Genista tenera(Fabaceae).This extract decreased a-glucosidase and glucose-6-phosphatase activities to 0.97 and 80.25%,respectively,being more effective than acarbose,and phlorizin,the positive controls,which reduced enzymes activities only to 17.39 and 96.06%.Once inflammation and oxidative stress are related to diabetic impairments,the anti-inflammatory activity of the extract was also evaluated,through its inhibitory activity over COX-1 enzyme(47.5%inhibition).Moreover,after induction of oxidative stress by UV radiation,the viability of irradiated rat liver hepatoma cells exposed to the extract was significantly higher(67.82%)than that promoted by ascorbic acid,the positive control(45.05%).In addition,the stability of the extract under gastrointestinal conditions was evaluated by HPLC–DAD-ESI–MS/MS.Flavonoid diglycosides were identified as the main constituents of the extract,and no alterations in the chemical composition nor in the antioxidant activity were observed after in vitro digestion with artificial gastric and pancreatic juices.展开更多
文摘Zhuo et al looked into the part of transmembrane 9 superfamily member 1(TM9SF1)in bladder cancer(BC),and evaluated if it can be used as a therapeutic target.They created a permanent BC cell line and tested the effects of TM9SF1 overexpression and suppression on BC cell growth,movement,invasion,and cell cycle advancement.Their results show that TM9SF1 can boost the growth,movement,and invasion of BC cells and their access into the G2/M stage of the cell cycle.This research gives a novel direction and concept for targeted therapy of BC.
基金This work was supported by a grant from the Scientific Research Foundation of Ministry of Public Health of PR China (No. 96-1-204).
文摘Objective: To study the expression of the inducible nitric oxide synthase (iNOS) gene and the effects of tumor necrosis factor-α(TNF-a) and interferon-γ(IFN-g)on proliferation of the continuous cultured human colon cancer cell line CCL229. Methods: Using the molecular and biochemical techniques and electron microscopy to analyze the expression of iNOS, production of NO and growth characteristics of human colon cancer cells. Results: cytokine treatment can induce expression of the iNOS gene and production of nitric oxide was significantly higher after treatment of CCL229 cells with TNF-αor IFN-γ. Treatment with either cytokine or a combination of both significantly increased levels of Malondialdehyde (MDA) over control. Furthermore, cytokine treatment increased the proliferation inhibition rate as assessed in vitro and decreased the cell proliferation index on flow cytometry. Electron microscopy showed that cells treated with cytokines had fewer pseudopodia or cell processes than control cells and that cytokine treated cells had dilatation of the mitochondria and endoplasmic reticulum and dilated vesicular or tubular cisternae. Conclusion: Our findings indicate that TNF-α and IFN-γ induce the expression of iNOS gene in CCL229 cells, which increases the production of nitric oxide, inhibits proliferation, causes lipid peroxidation, and results in ultrastructural changes.
文摘The undecapeptide substance P(SP) was shown to be intimately involved in both the structural and functional aspects of the anterior pituitary. Yet, in addition to its influences on hormonal secretion, SP may well possess more actions in this master gland. The present study was ftherefore aimed to investigate the effect of SP on the proliferation of rat anterior pituitary cells in primary culture. It was found that SP could dose-dependently increase the ineorporation of tritiated thymidine (3H-TdR)into cultured anterior pituitary cells. Other mammalian tachykinins such as neurokinin A and neurokinin B had similar effect but to varying degrees. The equipotent analogue of SP, Norleucine(11) -SP(Nle(11)-SP), also acted likewise, with its action antagonizable by spantide, a SP receptor blocker. To further characterize the nature of cells responsive to the challenge of SP, immunocytochemical staining against S-100 protein and some adenohypophyseal hormones was performed alone or plus autoradiography The results showed that the percentage of S-100 proteinimmunoreactive cells was apparently elevated by the addtion of Nle(11)-SP for 48 h, which indicates a preferential proliferation of folliculo-stellate cells under the regime. This was confirmed by increases in immunocytochemical or autoradiographical labelling indices of anterior pituitary cells treated similarly. Taken togethbr, these results reveal that the trophic action of SP observed previously in other tissues is also present at least in cultured rat anterior pituitary cells, with responding cells being predominantly folliculo-stellate cells as typified by S-100 proteinimmunoreactivity. Therefore, an intra-pituitary trophic action of SP in vivo could be anticipated.
基金supported by a grant from the National Natural Sciences Foundation of China (No. 30570568)
文摘Astrocytes play a major role in the reactive processes in response to neuronal injuries in the brain. Excessive gliosis is detrimental and can contribute to neuronal damage. CDS1 (TAPA), a member of the tetraspanin family of proteins, is upregulated by astrocytes after traumatic injury to the rat central nervous system (CNS). To further understand the role of CD81 in the inhibition of astrocytes, we analyzed the effects of a CD81 antibody, on cultured rat astrocytes. The results indicated that the effect worked in a dose-dependent manner with certain dosage range. It, however, reached a dosage equilibrium at a high dosage. Furthermore, anti-CD81 antibody remarkably inhibited the proliferation of astrocytes after incubation with astrocytes for different periods of time and the effect presented a time-dependent fashion. However, anti-CDS1 antibody substantially inhibited the proliferation of astrocytes at low density and middle density but slightly inhibited the proliferation of as- trocytes at high density, suggesting that the effect was positively correlated with the proliferative ability of astrocytes. Finally, the cell cycle of astrocytes exposured to anti-CD81 antibody was arrested in S phase at the initial stage and at G0/GI phase over time. These findings indicated that CD81 exert significant inhibitory effect, dose-dependently and time-dependently, on the proliferation of astrocytes and the effect is positively correlated with the proliferative capability of astrocytes.
基金supported by Jiangsu Province Natural Science Foundation of China(js2817298)
文摘Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequencing,and then cloned into the eukaryotic expression vector pIRES2-EGFP,which encodes green fluorescent protein(GFP),to construct p1RES2-EGFP-NDRG2 plasmid.OS-RC-2 cells with NDRG2 negative expression were transfected with p1RES2-EGFP-NDRG2 plasmid.The growth of transfected OS-RC-2 cells was observed under light and fluorescence microscopes.After colony-forming cell assays,cell proliferation detection and MTT assays,the growth curves of cells in each group were plotted to investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of OS-RC-2 cells.Cell cycle was determined by flow cytometry.Confocal laser scanning microscopy showed that NDRG2 protein was specifically located on subcellular organelle.Results:A eukaryotic expression vector p1RES2-EGFP-NDRG2 was successfully constructed.After NDRG2 transfection,the growth of OS-RC-2 cells was inhibited.Flow cytometry showed that cells were arrested in S phase but the peak of cell apoptosis was not present,and confocal laser scanning microscopy showed that NDRG2 protein was located in mitochondrion.Conclusions:NDRG2 can significantly inhibit the proliferation of OS-RC-2 cells in vitro and its protein is specifically expressed in the mitochondrion.
文摘The effects of bleomycin A5 (BLM A5) alone and combined with calmodulin inhibitor N-(4-aminobutyl)-5-chloro-2-naphthalene sulfonamide (W-13) on the proliferation on S-180 cells in vitro were studied. IC50 of BLM used alone for the cells was about 2.63 μg/ml, but it was reduced to 1/3.8 and 1/9.5 of 2.63 μg/ml when plus W-13 1, 5 μg/ml respectively. The results indicated that nontoxic doses of W-13 enhanced the hinibition of cell proliferation under the condition of BLM 0.5 - 2.5 μg/ ml. In colony forming test, the survival fraction of S-180 cells treated with BLM plus W-13 was decreased to 1/87 - 240 of that of the cells treated with BLM alone. The results suggest that W-13 can enhance antitumor activity of BLM in vitro and may be used as an synergist of BLM A5 in vivo.
基金Supported by Higher Education Commission,Islamabad,Pakistan grant,No.20-17590/NRPU/R&D/HEC/20212021.
文摘BACKGROUND Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages.In humans,their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs.Studies suggested that mesenchymal stem cells(MSCs),necessary for repair and regeneration via transplantation,require doses ranging from 10 to 400 million cells.Furthermore,the limited expansion of MSCs restricts their therapeutic application.AIM To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.METHODS Human umbilical cord(hUC)tissue derived MSCs were obtained and re-cultured.These cultured cells were subjected to the following evaluation pro-cedures:Immunophenotyping,immunocytochemical staining,trilineage differentiation,population doubling time and number,gene expression markers for proliferation,cell cycle progression,senescence-associatedβ-galactosidase assay,human telomerase reverse transcriptase(hTERT)expression,mycoplasma,cytomegalovirus and endotoxin detection.RESULTS Analysis of pluripotent gene markers Oct4,Sox2,and Nanog in recultured hUC-MSC revealed no significant differences.The immunophenotypic markers CD90,CD73,CD105,CD44,vimentin,CD29,Stro-1,and Lin28 were positively expressed by these recultured expanded MSCs,and were found negative for CD34,CD11b,CD19,CD45,and HLA-DR.The recultured hUC-MSC population continued to expand through passage 15.Proliferative gene expression of Pax6,BMP2,and TGFb1 showed no significant variation between recultured hUC-MSC groups.Nevertheless,a significant increase(P<0.001)in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs.Cellular senescence markers(hTERT expression andβ-galactosidase activity)did not show any negative effect on recultured hUC-MSCs.Additionally,quality control assessments consistently confirmed the absence of mycoplasma,cytomegalovirus,and endotoxin contamination.CONCLUSION This study proposes the development of a novel protocol for efficiently expanding stem cell population.This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies.
文摘A study was done to produce a rapid in vitro propagation of three strawberry genotypes and tested in the field under Bangladeshi circumstances. Festival, RABI-3, and Neho strawberry genotypes’ runner tips were cultivated in vitro to induce root induction and multiple shoot proliferation. MS (Murashige and Skoog) media that were basally containing three different concentrations at 1.0, 1.5, and 2.0 of BA (6-benzyl adenine), KIN (6-furfuryl amino purine), or GA<sub>3</sub> (gibberellic acid) at 0.5 mg/L increasing tips of the runner was attained. The culture grew on the medium provided with 1.5 mg/L 6-benzyl adenine and 0.5 mg/L 6-furfuryl amino acid to increase shoot at the best level. Micro-cuttings were rooted on MS media at half strength combined with 0.5 mg/L - 1.5 mg/L IBA (indole butyric acid) or IAA (indole acetic acid). IBA attained 4 - 9 roots and 91% - 96% rooting at 1.0 mg/L. The resulting plantlets grew into hardy plants and took root in the earth. The genotype festival had the highest response rate, followed by RABI-3 and Neho.
文摘BACKGROUND Zinc(Zn)is the second most abundant trace element after Fe,present in the human body.It is frequently reported in association with cell growth and proliferation,and its deficiency is considered to be a major disease contributing factor.AIM To determine the effect of Zn on in vitro growth and proliferation of human umbilical cord(hUC)-derived mesenchymal stem cells(MSCs).METHODS hUC-MSCs were isolated from human umbilical cord tissue and characterized based on immunocytochemistry,immunophenotyping,and tri-lineage differentiation.The impact of Zn on cytotoxicity and proliferation was determined by MTT and Alamar blue assay.To determine the effect of Zn on population doubling time(PDT),hUC-MSCs were cultured in media with and without Zn for several passages.An in vitro scratch assay was performed to analyze the effect of Zn on the wound healing and migration capability of hUC-MSCs.A cell adhesion assay was used to test the surface adhesiveness of hUC-MSCs.Transcriptional analysis of genes involved in the cell cycle,proliferation,migration,and selfrenewal of hUC-MSCs was performed by quantitative real-time polymerase chain reaction.The protein expression of Lin28,a pluripotency marker,was analyzed by immunocytochemistry.RESULTS Zn at lower concentrations enhanced the rate of proliferation but at higher concentrations(>100μM),showed concentration dependent cytotoxicity in hUC-MSCs.hUC-MSCs treated with Zn exhibited a significantly greater healing and migration rate compared to untreated cells.Zn also increased the cell adhesion rate,and colony forming efficiency(CFE).In addition,Zn upregulated the expression of genes involved in the cell cycle(CDC20,CDK1,CCNA2,CDCA2),proliferation(transforming growth factorβ1,GDF5,hypoxia-inducible factor 1α),migration(CXCR4,VCAM1,VEGF-A),and self-renewal(OCT4,SOX2,NANOG)of hUC-MSCs.Expression of Lin28 protein was significantly increased in cells treated with Zn.CONCLUSION Our findings suggest that zinc enhances the proliferation rate of hUC-MSCs decreasing the PDT,and maintaining the CFE.Zn also enhances the cell adhesion,migration,and self-renewal of hUC-MSCs.These results highlight the essential role of Zn in cell growth and development.
基金Supported by Major Project on Health Care and Innovation of Guangzhou,China(No.201400000003-3)Science and Technology Planning Project of Guangdong Province,China(No.2015A020212026)
文摘AIM: To explore whether resveratrol (Res) can inhibit human retinal pigment epithelial cell (ARPE-19 cell) proliferation and migration, and to research the molecular mechanisms.METHODS: ARPE-19 cells were pretreated with various concentrations at 0, 50, 100, 150, 200 and 300 μmol/L of Res, and with 0 μmol/L Res as the control for 24, 48 and 72h. The cell proliferation, apoptosis and migration were measured with cell counting kit-8 (CCK-8), flow cytometry, and wound-healing and Transwell assays, respectively. The expression of proliferating cell nuclear antigen (PCNA), P21 and P27, as well as matrix metalloproteinase-9 (MMP-9) and p38 mitogen-activated protein kinases (p38MAPK) was identified by Western blot.RESULTS: Cell proliferation was effectively inhibited by Res (P〈0.05). When pretreated with Res, cells arrested in S-phase increased remarkably (P〈0.05), but the apoptosis ratios showed no significant difference between the treatment and control groups (P〉0.05). Cell migration was suppressed by Res both in wound-healing assay and Transwell migration assay (P〈0.05). Decreases of PCNA, MMP-9 and p38MAPK, as well as increases of P21 and P27 were detected by Western blot (P〈0.05). CONCLUSION: Res can inhibit APRE-19 cell proliferation and migration in a concentration-dependent manner with up-regulation of the expression of P21 and P27, and down-regulation of PCNA, MMP-9 and p38MAPK.
基金the Hubei Province Nature Science Foundation, No. 2005ABA121
文摘AIM: To evaluate the effects of adenovirus-mediated gene transfer of RhoA siRNA and RhoC siRNA on proliferation and invasion of SGC7901 cells by Rho/ PI3K/Akt pathway. METHODS: Plasmid of RhoA siRNA and RhoC siRNA were constructed and transfected into SGC7901 cells. siRNA and LY294002 (PI3K inhibitor) were designed as the control group. The mRNA and protein expressions of RhoA and RhoC were respectively detected with RT-PCR and western blotting. In order to fi nd out the changes of proliferation and invasion power of SGC7901 cell lines, we analyzed the data by MTT, Boyden chamber and evaluated apoptosis of cell with flow cytometry. We treated BALB /C nude mice with RhoA and RhoC-siRNA, and tumor control rate (%) in nude mice was calculated. RESULTS: RhoA and RhoC siRNA transfections specifically down-regulated the corresponding mRNA and protein levels in SGC7901 Cells.The experiment of permeated artificial basal membrane showed that the invasion power of SGC7901 cell lines are on the decline after treatment of Ad-RhoA and RhoC-siRNA (12.64 ± 3.27 vs 87.38 ± 17.38, P < 0.05). The values of 490 nm wavelength light absorption were different in the five groups. The number of alive cells in the group of RhoA and RhoC-siRNA was lower than others in the 6th d (0.71 ± 0.01 vs 3.82 ± 0,11 P < 0.05). The apoptosis rate of transfected RhoA and RhoC-siRNA group with FACS were 19.07% ± 1.78 and there were significant differences between treated and control groups (19.07 ± 1.78% vs 1.23 ± 0.11%, P < 0.01). The tumor transplantation experiment in BALB/C nude mice showed intratumoralinjection of RhoA or RhoC siRNA can inhibit tumor growth. CONCLUSION: RhoA and RhoC siRNA gene therapy mediated by adenovirus may be useful for inhibiting growth and invasion of SGC7901 through a PI3K/Akt pathway. These results provide a novel therapeutic target in preventing gastric cancer cell invasion and metastasis.
基金supported by a grant from the Natural Sciences Foundation of Hubei Province,China(No.2008CDA055)
文摘The effects of DK2,a peroxisome proliferator-activated receptor γ agonist,on cultured human pterygium fibroblasts (HPFs) in virto were studied.The HPFs were incubated with 0-200 μmol/L DK2 for 12-72 h.The MTT method was used to assay the bio-activity of DK2 at different doses and time.The cytotoxic effect of DK2 was measured by LDH release assay.The cell cycle distribution and apoptosis were flow cytometrically detected.The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by real-time PCR (RT-PCR) and Western blotting.The results showed that administration of 1-75 μmol/L DK2 for 12-72 h could significantly inhibit HPF proliferation in a dose-and time-dependent manner.DK2-treated cells did not release significant amount of LDH as compared with rosiglitazone-treated cells.After treatment with DK2 at concentrations of 15,25 μmol/L for 24 h,the number of HPFs in G 0 /G 1 phase was significantly increased while that in S phase was significantly decreased (P【0.05),leading to arrest at G 0 /G 1 phase.The apoptosis rates of HPF cells in drug-treated groups were significantly higher than the rate of control group (P【0.05).At the dosage range between 15-25 μmol/L,DK2 could inhibit the expression of PCNA mRNA and protein in HPFs in a dose-dependent fashion (P【0.05).It was concluded that PPARγ agonist can significantly inhibit HPF proliferation,resulting in the arrest at G 0 /G 1 phase,induce the apoptosis of HPFs,and suppress the synthesis of PCNA,in dose-and time-dependent manners.
基金supported by a grant from the National Key Technology Research and Development Program of the Ministry of Science and Technology of China,No.2014BAI01B00
文摘Neural stem cells promote neuronal regeneration and repair of brain tissue after injury,but have limited resources and proliferative ability in vivo.We hypothesized that nerve growth factor would promote in vitro proliferation of neural stem cells derived from the tree shrews,a primate-like mammal that has been proposed as an alternative to primates in biomedical translational research.We cultured neural stem cells from the hippocampus of tree shrews at embryonic day 38,and added nerve growth factor(100 μg/L) to the culture medium.Neural stem cells from the hippocampus of tree shrews cultured without nerve growth factor were used as controls.After 3 days,fluorescence microscopy after DAPI and nestin staining revealed that the number of neurospheres and DAPI/nestin-positive cells was markedly greater in the nerve growth factor-treated cells than in control cells.These findings demonstrate that nerve growth factor promotes the proliferation of neural stem cells derived from tree shrews.
文摘The effect of 1,25-dihydroxyvitamin D<sub>3</sub> [1,25(OH)<sub>2</sub>D<sub>3</sub>] and13-cis-retinoic acid(RA)on the proliferation of a novel human megakaryoblasticleukemia cell line(HIMeg)was investigated.At the concentration of 10<sup>9</sup> 10<sup>6</sup>mol/L,1,25(OH)<sub>2</sub>D<sub>3</sub> and RA showed significant inhibition of the proliferation of themegakaryoblastic leukemic cells,which was demonstrated by the count of survival cells,incorporation of<sup>3</sup>H-TdR and<sup>3</sup>H-UR,and cloning efficiency in dose-dependent and time-dependent manners.The results can further explain the mechanism of differentiation-inducing agents and the effect of 1 ,25(OH)<sub>2</sub>D<sub>3</sub> on myelofibrosis.It is possible for1,25(OH)<sub>2</sub>D<sub>3</sub> and RA to be used to treat malignant megakaryocytic diseases.
基金This project was supported by the Teaching and Research Award Program for Outstanding Young Teachers in Higher Education Institutions of MOE of China
文摘Summary: The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx Hep-2 in vitro and their corresponding mechanisms were investigated. Hep-2 cells were cultured with two kinds of selective cyclooxygenase-2 inhibitors (Sc-58125 and Celecoxib) at various concentrations for 24 h. Morphological changes were observed under the phase microscopy and the growth suppression was detected by using MTT colorimetric assay. Apoptotic DNA fragments were observed by agarose gel electrophoresis, and the cell cycle and apoptotic rate were detected by flow cytometry (FCM) respectively. Hep-2 cells became rounded and detached from the culture dish after being treated with Celecoxib for 24 h, however, they remained morphologically unchanged with Sc-58125. Sc-58125 could increase G 2 phase cells, whereas, Celecoxib rose G 1 phase cells. Both of the two effects were dose-dependent. Moreover, the Hep-2 cells cultured with 50 μmol/L and 100 μmol/L Celecoxib showed obvious apoptosis, with the nuclear DNA of cells exhibiting characteristic DNA ladder. So Sc-58125 could inhibit the proliferation of Hep-2 cells by altering the G 2 phase cells. However, Celecoxib had the same effect by changing the G 1 phase cells and inducing apoptosis at higher concentration.
文摘Acacia karroo Hayne is an arbor species widely distributed in South Africa with the characteristics of fast growth and drought resistance. The species was introduced to China recently. In vitro culture is an effective method to rapidly produce plants and a strategy to minimize somaclonal variation among regenerated plants. Browning, however, is a problem in establishing the in vitro culture system. The present study diminished the problem by selecting explants, using different browning inhibitors and chilling treatment. Results showed that the use of embryos as explants reduced the browning ratio to 46.7%, whilst stem segment explants were browned up to 56.7%. The adventitious buds were successfully induced in the modified tissue culture medium supplemented with 5.0 mg·L^-1 6-BA and 0.1 mg·L^-1 NAA. The proliferation coefficient of adventitious buds is 2.8.
文摘Objective:To observe the effects of platelet-rich plasma(PRP)on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue.Methods:Fibroblasts were separated from human chronic refractory wound granulation tissue and then were identified.The obtained fibroblasts were divided into fetal bovine serum(FBS)group,hydrogel group and PRP group,and the three groups were cultured with culture mediums containing FBS,hydrogel and PRP respectively,in order to observe the growth of fibroblasts.The wound scratch assay was used to observe the migration of fibroblasts.Results:PRP group had more fibroblasts than FBS group and hydrogel group since Day 5 of culture,and exhibited greater fibroblast scratch migration area than FBS group on 48 h and 72 h of wound scratch assay(all p<.05).Conclusions:Compared with FBS,human fibroblasts cultured by PRP can more effectively promote the proliferation and migration of fibroblasts.
基金The authors gratefully acknowledge Fundacāo para a Ciência e a Tecnologia for financial support of the projects PTDC/QUI/67165/2006 and PEst-OE/QUI/UI0612/2013The authors also thank the European Commission for approval of the INOVAFUNAGEING commitment and the support of the project“PERsonalised ICT supported Service for Independent Living and Active Ageing”,FP7-ICT-2013-10,Project Nr.610359,2013-2016The collaboration of the staff from the Herbarium of Jardim Botanico da Madeira,for providing plant material,is also acknowledged.
文摘The inhibition of a-glucosidase and glucose-6-phosphatase,two enzymes involved in the carbohydrate metabolism,is an important target to control glycaemia on individuals with type 2 diabetes.In this work we report for the first time the inhibition of both enzymes by the antihyperglycemic n-butanol extract from Genista tenera(Fabaceae).This extract decreased a-glucosidase and glucose-6-phosphatase activities to 0.97 and 80.25%,respectively,being more effective than acarbose,and phlorizin,the positive controls,which reduced enzymes activities only to 17.39 and 96.06%.Once inflammation and oxidative stress are related to diabetic impairments,the anti-inflammatory activity of the extract was also evaluated,through its inhibitory activity over COX-1 enzyme(47.5%inhibition).Moreover,after induction of oxidative stress by UV radiation,the viability of irradiated rat liver hepatoma cells exposed to the extract was significantly higher(67.82%)than that promoted by ascorbic acid,the positive control(45.05%).In addition,the stability of the extract under gastrointestinal conditions was evaluated by HPLC–DAD-ESI–MS/MS.Flavonoid diglycosides were identified as the main constituents of the extract,and no alterations in the chemical composition nor in the antioxidant activity were observed after in vitro digestion with artificial gastric and pancreatic juices.