Using electroacupuncture and moxibustion to treat peripheral nerve injury is highly efficient with low side effects. However, the electroacupuncture-and moxibustion-based mechanisms underlying nerve repair are still u...Using electroacupuncture and moxibustion to treat peripheral nerve injury is highly efficient with low side effects. However, the electroacupuncture-and moxibustion-based mechanisms underlying nerve repair are still unclear. Here, in vivo and in vitro experiments uncovered one mechanism through which electroacupuncture and moxibustion affect regeneration after peripheral nerve injury. We first established rat models of sciatic nerve injury using neurotomy. Rats were treated with electroacupuncture or moxibustion at acupoints Huantiao （GB30） and Zusanli （ST36）. Each treatment lasted 15 minutes, and treatments were given six times a week for 4 consecutive weeks. Behavioral testing was used to determine the sciatic functional index. We used electrophysiological detection to measure sciatic nerve conduction velocity and performed hematoxylin-eosin staining to determine any changes in the gastrocnemius muscle. We used immunohistochemistry to observe changes in the expression of S100—a specific marker for Schwann cells—and an enzyme-linked immunosorbent assay to detect serum level of nerve growth factor. Results showed that compared with the model-only group, sciatic functional index, recovery rate of conduction velocity, diameter recovery of the gastrocnemius muscle fibers, number of S100-immunoreactive cells,and level of nerve growth factor were greater in the electroacupuncture and moxibustion groups. The efficacy did not differ between treatment groups. The serum from treated rats was collected and used to stimulate Schwann cells cultured in vitro. Results showed that the viability of Schwann cells was much higher in the treatment groups than in the model group at 3 and 5 days after treatment. These findings indicate that electroacupuncture and moxibustion promoted nerve regeneration and functional recovery; its mechanism might be associated with the enhancement of Schwann cell proliferation and upregulation of nerve growth factor.展开更多
Objective： To observe the effect of Bushen Tiaochong Recipe （补肾调冲方, BSTCR） on rats＇ ovarian granulosa cell （GC） proliferation, steroidogenesis and follicle-stimulating hormone receptor （FSHR）, and insulin...Objective： To observe the effect of Bushen Tiaochong Recipe （补肾调冲方, BSTCR） on rats＇ ovarian granulosa cell （GC） proliferation, steroidogenesis and follicle-stimulating hormone receptor （FSHR）, and insulin-like growth factor-1 （IGF-1） mRNA expression using serum pharmacological method. Methods： Rats＇ GCs were incubated with 10% blank serum （as negative control group）, follicle- stimulating hormone （FSH）-containing serum （S-FSH, as positive control group）, or BSTCR （in different dosages） containing serum （S-BSTCR, as the BSTCR groups） for 48 h. ^3H-TdR incorporation was then performed; DNA was measured to analyze the distribution of GCs in the cell cycle and their proliferation index （PI） using a flow cytometer; estradiol （E2） and progesterone （P） content in the culture fluid were examined by radioimmunoassay; and levels of FSHR and IGF-1 mRNA expression in GCs were measured by real-time RT-PCR. Results： A dose-dependent increase of ^3H-TdR incorporation in GC was shown in the BSTCR groups. Cells in Go/G1 phase had markedly less, while those in S phase had a significantly higher increase in the BSTCR groups compared with the negative control group. A high value of PI was also shown in the BSTCR groups, especially in the high dose group where the influence of cell proliferation was stronger than that in the positive control group. The levels of E2 and P in the BSTCR groups of all dosages were significantly higher than those in the negative control group, and did not show any significant difference compared with those in the positive control group. Levels of FSHR and IGF-1 mRNA expression in the BSTCR groups increased in a dose-dependent manner at levels higher than those in the negative control group. Conclusion： S-BSTCR can obviously stimulate the proliferation and steroidogenesis of ovarian GCs.It is speculated that BSTCR could play a regulatory action on ovarian function through two different pathways of endocrine and autocrine by promoting FSHR and IGF-1 mRNA expression.展开更多
BACKGROUND： How to induce endogenous neural stem cells （NSCs） to differentiate into needed neural cell types is a hot spot of current researches. OBJECTIVE： To compare differences between fetal bovine serum and Ch...BACKGROUND： How to induce endogenous neural stem cells （NSCs） to differentiate into needed neural cell types is a hot spot of current researches. OBJECTIVE： To compare differences between fetal bovine serum and Chinese herbal formula Naoluoxintong serum supplementation for inducing proliferation and differentiation in rat embryonic NSCs. DESIGN, TIME AND SETTING： An in vitro, serum pharmacology, comparative, observation study was performed from March to September in 2008 at the Laboratory of Neurodegenerative Diseases, College of Life Science in University of Science and Technology of China, the Key Laboratory Breeding Base of Acupuncture Foundation and Technology in Anhui University of Traditional Chinese Medicine, the Anhui Province Key Laboratory of R ＆ D of Chinese Medicine, and at the Level 3 Laboratory of Molecular Biology of the State Administration of Traditional Chinese Medicine. MATERIALS： The Chinese herbal formula Naoluoxintong was produced by Radix Astragali, Radix Notoginseng, Rhizoma Chuanxiong, Scolopendra at Anhui University of Traditional Chinese Medicine. Mouse anti-rat nestin, gliat fibrillary acidic protein, and galactocerebroside monoclonal antibodies, as well as rabbit anti-neuron-specific enolase polyclonal antibody were produced by Chemicon, Billerica, MA, USA. METHODS： Wistar rats aged 3 months were intragastrically infused with Naoluoxintong. Wistar rat embryonic NSCs （passage 8） were induced to proliferate and differentiate using 10% fetal bovine serum, 10% Naoluoxintong serum, and 10% rat serum. MAIN OUTCOME MEASURES： Phenotypic changes in cultured cells were detected using phase contrast microscopy, and cell proliferation and differentiation were observed using immunofluorescence staining. RESULTS： Proliferation and differentiation of embryonic NSCs was induced by three different types of blood serum. Although the differentiation time course with Nao/uoxintongserum was later than with the other two methods, the differentiated cells were morphologically similar to mature neurons to a greater extent. CONCLUSION： Nao/uoxintong serum supplementation induced differentiation of NSCs into neuronal-like cells and stimulated neuronal maturation.展开更多
Glutamate application is an established method of inducing PC12 cell injury. PC12 cells were cultured with serum containing Tongqiaohuoxue decoction consisting of moschus, Carthamus tinctonus, Rhizoma chuanxiong, Seme...Glutamate application is an established method of inducing PC12 cell injury. PC12 cells were cultured with serum containing Tongqiaohuoxue decoction consisting of moschus, Carthamus tinctonus, Rhizoma chuanxiong, Semen pruni persicae, and Radix Paeoniae Rubra. After 24 hours of co-cultivation, glutamate （12.5 mM） was added to the culture medium. We found that serum containing Tongqiaohuoxue decoction prevented the increase in reactive oxygen species, and the decreases in superoxide dismutase and Na＋-K＋-ATPase activity, induced by glutamate. It also reduced the concentration of malondialdehyde, enhanced the mitochondrial transmembrane potential, inhibited the elevation of cellular calcium, and decreased phosphorylation of calmodulin-dependent protein kinase I1. Thus, serum containing Tongqiaohuoxue decoction had protective effects on cell proliferation and membrane permeability in glutamate-injured PC12 cells.展开更多
Background:Abnormal proliferation of T cells plays an essential role in the pathogenesis of Systemic lupus erythematosus(SLE).The pharmaceutical effect of Hedyotis Diffusa Willd(HDW)on SLE has been investigated previo...Background:Abnormal proliferation of T cells plays an essential role in the pathogenesis of Systemic lupus erythematosus(SLE).The pharmaceutical effect of Hedyotis Diffusa Willd(HDW)on SLE has been investigated previously.Nevertheless,the biomedical mechanism is still left unclear.Objective:This study has been arranged to evaluate the therapeutic effect of the ethyl acetate fraction of HDW(EAHDW)on lupus mice and explore the potential therapeutic mechanism.Methods:EAHDW was prepared with 80%ethanol reflex extraction followed by successive extraction,and ana-lyzed with HPLC and UPLC-Q/TOP-MS.The potential targets and STAT3 affinity regulators were predicted with network pharmacology.The pharmaceutic effect of EAHDW was studied with MRL/lpr mice.Cytokines and au-toantibodies were quantified with ELISA assays.The pathological damage of glomerulus and STAT3 expression in the kidney was detected with histochemical and immunohistochemical techniques.The cell cycle properties in cell proliferation were identified with the flow cytometry.The western blot and dual-Luciferase reporter assay were applied to evaluate translational and transcriptional activity of STAT3,respectively.Results:In this study,the extraction ratio of EAHDW was 2.7±1%,in which 19 ingredients were identified.Network pharmacological analysis showed that the target genes of EAHDW were highly focused on influencing the abnormal T cell proliferation in SLE.EAHDW showed the beneficial effects on pathological changes and STAT3 expression in the glomerulus of lupus mice,and the levels of cytokines and autoantibodies in serum.In cytological study,EAHDW treatment attenuated the transcription and phosphorylation of STAT3,which inhibited T cell proliferation by prolonged S-phase of the cell cycle.A total of 5 compounds in EAHDW exhibited high docking affinity to the DNA-binding site of STAT3.Conclusion:EAHDW could reduce the inflammatory response and inhibit the proliferation of T cells by interfering with the STAT3 signaling pathway,thereby playing a therapeutic effect on SLE.展开更多
BACKGROUND Pachymic acid(PA)is derived from Poria cocos.PA has a variety of pharmacological and inhibitory effects on various tumors.However,the mechanism of action of PA in gastric cancer(GC)remains unclear.AIM To in...BACKGROUND Pachymic acid(PA)is derived from Poria cocos.PA has a variety of pharmacological and inhibitory effects on various tumors.However,the mechanism of action of PA in gastric cancer(GC)remains unclear.AIM To investigate the mechanism of PA in treating GC via the combination of network pharmacology and experimental verification.METHODS The GeneCards and OMIM databases were used to derive the GC targets,while the Pharm Mapper database provided the PA targets.Utilizing the STRING database,a protein-protein interaction network was constructed and core targets were screened.The analyses of Gene Ontology,Kyoto Encyclopedia of Genes and Genomes(KEGG),and gene set enrichment analysis were conducted,and molecular docking and clinical correlation analyses were performed on the core targets.Ultimately,the network pharmacology findings were validated through in vitro cell assays,encompassing assessments of cell viability,apoptosis,cell cycle,cloning,and western blot analysis.RESULTS According to network pharmacology analysis,the core targets were screened,and the PI3K/AKT signaling pathway is likely to be the mechanism by which PA effectively treats GC,according to KEGG enrichment analysis.The experimental findings showed that PA could control PI3K/AKT signaling to prevent GC cell proliferation,induce apoptosis,and pause the cell cycle.CONCLUSION Network pharmacology demonstrated that PA could treat GC by controlling a variety of signaling pathways and acting on a variety of targets.This has also been supported by in vitro cell studies,which serve as benchmarks for further research.展开更多
The functional formula diet AHP, containing polysaccharides from Auricularia auricular, polyphenolic compounds from Hawthorn (Crataegus) and Pueraria radix, has been recently developed as a dietary intervention agains...The functional formula diet AHP, containing polysaccharides from Auricularia auricular, polyphenolic compounds from Hawthorn (Crataegus) and Pueraria radix, has been recently developed as a dietary intervention against dyslipidemia in our previous study. In the present study, its antioxidant activities and protective effects against proliferation of vascular smooth muscle cells (VSMCs) were investigated. AHP possessed the potent radical-scavenging effects against hydroxyl and superoxide radicals, and also the inhibitory effects against peroxidation of low density lipoprotein induced by Cu2+ in vitro. The protective effects of AHP against proliferation of VSMCs were evaluated through the methodology of serum pharmacology. The serum containing AHP significantly inhibited the proliferation of VSMCs induced by oxidized low density lipoprotein in a time- and dose-dependent manner, and also promoted the nitric oxide production of VSMCs. Our study indicated that this functional formula diet would be a potent alternative as a functional diet to prevent atherosclerosis at early stage.展开更多
[Objectives]To observe the effect of drug-containing serum from different extracts of Phyllanthus reticulatus leaf on the proliferation of human gastric cancer SGC-7901 cells and the effect of in vivo on the life exte...[Objectives]To observe the effect of drug-containing serum from different extracts of Phyllanthus reticulatus leaf on the proliferation of human gastric cancer SGC-7901 cells and the effect of in vivo on the life extension rate of mice with H22 ascites tumor,and to investigate the effects of chemical components such as Corilagin,ethyl gallate,ethyl brevifolincarboxylate,and gallic acid on the proliferation of human gastric cancer SGC-7901 cells.[Methods]MTT method was used to observe the inhibitory effect of drug-containing serum of different extracts of P.reticulatus leaf on human gastric cancer SGC-7901 cells in vitro to determine its active site.The active site was used as the research object to establish a Kunming mouse ascites tumor model,to investigate the effect on the life extension rate of mice with H22 ascites tumor,to further separate the monomer components from the effective fraction and investigate its effect on human gastric cancer SGC-7901 cells.[Results]Compared with the 10%blank serum control group,10%drug-containing serum of ethyl acetate and n-butanol in P.reticulatus leaf had significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and the inhibition rate was 34.99%and 28.68%,respectively(P<0.05).Compared with the blank group,the survival time of the high dose group of ethyl acetate in P.reticulatus leaf was significant(P<0.05).Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester had a significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,with IC 50 of 26.52,70.45,and 158.86μg/mL,respectively.Ethyl gallate had no inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and its IC 50 was 251.96μg/mL.[Conclusions]The drug-containing serum of ethyl acetate and n-butanol extract of P.reticulatus can inhibit the proliferation of human gastric cancer SGC-7901 cells.Ethyl acetate of P.reticulatus leaves can increase the life extension rate of mice with H22 ascites tumor.Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester isolated from its active site are the material basis for inhibiting the proliferation of human gastric cancer SGC-7901 cells.展开更多
Objective： To investigate the effect of Jinmaitong （筋脉通,JMT） serum on the proliferation of rat Schwann cells （SCs） primarily cultured in high glucose medium. Method： SOs were primarily cultured in Dulbecco＇s...Objective： To investigate the effect of Jinmaitong （筋脉通,JMT） serum on the proliferation of rat Schwann cells （SCs） primarily cultured in high glucose medium. Method： SOs were primarily cultured in Dulbecco＇s minmum essential medium （DMEM control）, 50 mmol/L glucose medium （50 mmol/L Glu）, 75 mmol/L glucose medium （75 mmol/L Glu）, as well as 50 mmol/L glucose medium, with different concentrations of JMT serum （undiluted, 1：2 diluted and 1：8 diluted） and Neurotropin （Ntp）, respectively. The proliferation of SCs under different conditions was detected by MTT. Result： SCs grew exuberantly in DMEM within 24-72 h, but slowed down at 96 h. The proliferation of SCs was inhibited in 50 mmol/L Glu and 75 mmol/L Glu after cultures of 48, 72 and 96 h, which showed that both were significantly different compared to the control group （P〈0.01）. The inhibition was more significant in 75 mmol/L Glu than in 50 mmol/L Glu （P〈0.05）. Spearman＇s rho analysis revealed that the proliferation of SCs had a negative correlation with the concentration of glucose （r=-0.471, P〈0.01）. Excluding the time factor, partial correlation showed similar results （r =-0.679, P〈0.01）. After 48 h, the proliferation of SCs increased significantly in JMT 1：2 and Ntp compared with 50 mmol/L Glu （control 0.437±0.019, 50 mmol/ L Glu 0.367±0.035, JMT1：2 0.426±0.024, Ntp 0.422±0.013; P〈0.01）, and there were no statistically significant differences among the JMT groups, the Ntp group and the control group （P〉0.05）. Conclusions： The proliferation of SCs was inhibited in high glucose medium, and the inhibition was reduced by different concentrations of JMT serum, especially at JMT1：2.展开更多
Background The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood （PB-MSCs） were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the f...Background The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood （PB-MSCs） were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair. Methods MSCs were mobilized into peripheral blood by granulocyte colony stimulating factor （G-CSF） and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension （20% 02 and 2% O2）, with or without 10% fetal bovine serum （FBS） conditions： 20% 02 and 10% FBS complete medium （normal medium, N）, 20% 02 and serum deprivation medium （D）, 2% 02 and 10% FBS complete medium （hypoxia, H）, 2% 02 and serum deprivation （HD）. Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling （TUNEL） staining. Results Spindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow （BM-MSCs）, and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II （MHC II）. They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia. Conclusions PB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.展开更多
基金supported by the National Natural Science Foundation of China,No.81373754,81102670
文摘Using electroacupuncture and moxibustion to treat peripheral nerve injury is highly efficient with low side effects. However, the electroacupuncture-and moxibustion-based mechanisms underlying nerve repair are still unclear. Here, in vivo and in vitro experiments uncovered one mechanism through which electroacupuncture and moxibustion affect regeneration after peripheral nerve injury. We first established rat models of sciatic nerve injury using neurotomy. Rats were treated with electroacupuncture or moxibustion at acupoints Huantiao （GB30） and Zusanli （ST36）. Each treatment lasted 15 minutes, and treatments were given six times a week for 4 consecutive weeks. Behavioral testing was used to determine the sciatic functional index. We used electrophysiological detection to measure sciatic nerve conduction velocity and performed hematoxylin-eosin staining to determine any changes in the gastrocnemius muscle. We used immunohistochemistry to observe changes in the expression of S100—a specific marker for Schwann cells—and an enzyme-linked immunosorbent assay to detect serum level of nerve growth factor. Results showed that compared with the model-only group, sciatic functional index, recovery rate of conduction velocity, diameter recovery of the gastrocnemius muscle fibers, number of S100-immunoreactive cells,and level of nerve growth factor were greater in the electroacupuncture and moxibustion groups. The efficacy did not differ between treatment groups. The serum from treated rats was collected and used to stimulate Schwann cells cultured in vitro. Results showed that the viability of Schwann cells was much higher in the treatment groups than in the model group at 3 and 5 days after treatment. These findings indicate that electroacupuncture and moxibustion promoted nerve regeneration and functional recovery; its mechanism might be associated with the enhancement of Schwann cell proliferation and upregulation of nerve growth factor.
基金Supported by Jiangsu Provincial Natural Science Foundation (No.BK2005006)
文摘Objective： To observe the effect of Bushen Tiaochong Recipe （补肾调冲方, BSTCR） on rats＇ ovarian granulosa cell （GC） proliferation, steroidogenesis and follicle-stimulating hormone receptor （FSHR）, and insulin-like growth factor-1 （IGF-1） mRNA expression using serum pharmacological method. Methods： Rats＇ GCs were incubated with 10% blank serum （as negative control group）, follicle- stimulating hormone （FSH）-containing serum （S-FSH, as positive control group）, or BSTCR （in different dosages） containing serum （S-BSTCR, as the BSTCR groups） for 48 h. ^3H-TdR incorporation was then performed; DNA was measured to analyze the distribution of GCs in the cell cycle and their proliferation index （PI） using a flow cytometer; estradiol （E2） and progesterone （P） content in the culture fluid were examined by radioimmunoassay; and levels of FSHR and IGF-1 mRNA expression in GCs were measured by real-time RT-PCR. Results： A dose-dependent increase of ^3H-TdR incorporation in GC was shown in the BSTCR groups. Cells in Go/G1 phase had markedly less, while those in S phase had a significantly higher increase in the BSTCR groups compared with the negative control group. A high value of PI was also shown in the BSTCR groups, especially in the high dose group where the influence of cell proliferation was stronger than that in the positive control group. The levels of E2 and P in the BSTCR groups of all dosages were significantly higher than those in the negative control group, and did not show any significant difference compared with those in the positive control group. Levels of FSHR and IGF-1 mRNA expression in the BSTCR groups increased in a dose-dependent manner at levels higher than those in the negative control group. Conclusion： S-BSTCR can obviously stimulate the proliferation and steroidogenesis of ovarian GCs.It is speculated that BSTCR could play a regulatory action on ovarian function through two different pathways of endocrine and autocrine by promoting FSHR and IGF-1 mRNA expression.
基金National Natural Science Foundation of China,No.30873293,30672592Natural Science Foundation of Anhui Province,No.070413125,050430904+1 种基金Dr.Yafang Lü Graduate Research Foundation of Beijing University of Chinese Medicine,No.2004Natural Science Research Fund of Education Department of Anhui Province,No.2006KJ382B
文摘BACKGROUND： How to induce endogenous neural stem cells （NSCs） to differentiate into needed neural cell types is a hot spot of current researches. OBJECTIVE： To compare differences between fetal bovine serum and Chinese herbal formula Naoluoxintong serum supplementation for inducing proliferation and differentiation in rat embryonic NSCs. DESIGN, TIME AND SETTING： An in vitro, serum pharmacology, comparative, observation study was performed from March to September in 2008 at the Laboratory of Neurodegenerative Diseases, College of Life Science in University of Science and Technology of China, the Key Laboratory Breeding Base of Acupuncture Foundation and Technology in Anhui University of Traditional Chinese Medicine, the Anhui Province Key Laboratory of R ＆ D of Chinese Medicine, and at the Level 3 Laboratory of Molecular Biology of the State Administration of Traditional Chinese Medicine. MATERIALS： The Chinese herbal formula Naoluoxintong was produced by Radix Astragali, Radix Notoginseng, Rhizoma Chuanxiong, Scolopendra at Anhui University of Traditional Chinese Medicine. Mouse anti-rat nestin, gliat fibrillary acidic protein, and galactocerebroside monoclonal antibodies, as well as rabbit anti-neuron-specific enolase polyclonal antibody were produced by Chemicon, Billerica, MA, USA. METHODS： Wistar rats aged 3 months were intragastrically infused with Naoluoxintong. Wistar rat embryonic NSCs （passage 8） were induced to proliferate and differentiate using 10% fetal bovine serum, 10% Naoluoxintong serum, and 10% rat serum. MAIN OUTCOME MEASURES： Phenotypic changes in cultured cells were detected using phase contrast microscopy, and cell proliferation and differentiation were observed using immunofluorescence staining. RESULTS： Proliferation and differentiation of embryonic NSCs was induced by three different types of blood serum. Although the differentiation time course with Nao/uoxintongserum was later than with the other two methods, the differentiated cells were morphologically similar to mature neurons to a greater extent. CONCLUSION： Nao/uoxintong serum supplementation induced differentiation of NSCs into neuronal-like cells and stimulated neuronal maturation.
基金supported by the National Natural Science Foundation of China, No. 30973979the Science-Technology Foundation for Excellent Young Scholar of Anhui Province China, No. 10040606Y17Kanion Innovation Fund of Traditional Chinese Medicine, No. KYCX2010007
文摘Glutamate application is an established method of inducing PC12 cell injury. PC12 cells were cultured with serum containing Tongqiaohuoxue decoction consisting of moschus, Carthamus tinctonus, Rhizoma chuanxiong, Semen pruni persicae, and Radix Paeoniae Rubra. After 24 hours of co-cultivation, glutamate （12.5 mM） was added to the culture medium. We found that serum containing Tongqiaohuoxue decoction prevented the increase in reactive oxygen species, and the decreases in superoxide dismutase and Na＋-K＋-ATPase activity, induced by glutamate. It also reduced the concentration of malondialdehyde, enhanced the mitochondrial transmembrane potential, inhibited the elevation of cellular calcium, and decreased phosphorylation of calmodulin-dependent protein kinase I1. Thus, serum containing Tongqiaohuoxue decoction had protective effects on cell proliferation and membrane permeability in glutamate-injured PC12 cells.
文摘Background:Abnormal proliferation of T cells plays an essential role in the pathogenesis of Systemic lupus erythematosus(SLE).The pharmaceutical effect of Hedyotis Diffusa Willd(HDW)on SLE has been investigated previously.Nevertheless,the biomedical mechanism is still left unclear.Objective:This study has been arranged to evaluate the therapeutic effect of the ethyl acetate fraction of HDW(EAHDW)on lupus mice and explore the potential therapeutic mechanism.Methods:EAHDW was prepared with 80%ethanol reflex extraction followed by successive extraction,and ana-lyzed with HPLC and UPLC-Q/TOP-MS.The potential targets and STAT3 affinity regulators were predicted with network pharmacology.The pharmaceutic effect of EAHDW was studied with MRL/lpr mice.Cytokines and au-toantibodies were quantified with ELISA assays.The pathological damage of glomerulus and STAT3 expression in the kidney was detected with histochemical and immunohistochemical techniques.The cell cycle properties in cell proliferation were identified with the flow cytometry.The western blot and dual-Luciferase reporter assay were applied to evaluate translational and transcriptional activity of STAT3,respectively.Results:In this study,the extraction ratio of EAHDW was 2.7±1%,in which 19 ingredients were identified.Network pharmacological analysis showed that the target genes of EAHDW were highly focused on influencing the abnormal T cell proliferation in SLE.EAHDW showed the beneficial effects on pathological changes and STAT3 expression in the glomerulus of lupus mice,and the levels of cytokines and autoantibodies in serum.In cytological study,EAHDW treatment attenuated the transcription and phosphorylation of STAT3,which inhibited T cell proliferation by prolonged S-phase of the cell cycle.A total of 5 compounds in EAHDW exhibited high docking affinity to the DNA-binding site of STAT3.Conclusion:EAHDW could reduce the inflammatory response and inhibit the proliferation of T cells by interfering with the STAT3 signaling pathway,thereby playing a therapeutic effect on SLE.
基金Supported by Ningxia Science and Technology Benefiting People Program,No.2022CMG03064National Natural Science Foundation of China,No.82260879Ningxia Natural Science Foundation,No.2022AAC03144 and 2022AAC02039.
文摘BACKGROUND Pachymic acid(PA)is derived from Poria cocos.PA has a variety of pharmacological and inhibitory effects on various tumors.However,the mechanism of action of PA in gastric cancer(GC)remains unclear.AIM To investigate the mechanism of PA in treating GC via the combination of network pharmacology and experimental verification.METHODS The GeneCards and OMIM databases were used to derive the GC targets,while the Pharm Mapper database provided the PA targets.Utilizing the STRING database,a protein-protein interaction network was constructed and core targets were screened.The analyses of Gene Ontology,Kyoto Encyclopedia of Genes and Genomes(KEGG),and gene set enrichment analysis were conducted,and molecular docking and clinical correlation analyses were performed on the core targets.Ultimately,the network pharmacology findings were validated through in vitro cell assays,encompassing assessments of cell viability,apoptosis,cell cycle,cloning,and western blot analysis.RESULTS According to network pharmacology analysis,the core targets were screened,and the PI3K/AKT signaling pathway is likely to be the mechanism by which PA effectively treats GC,according to KEGG enrichment analysis.The experimental findings showed that PA could control PI3K/AKT signaling to prevent GC cell proliferation,induce apoptosis,and pause the cell cycle.CONCLUSION Network pharmacology demonstrated that PA could treat GC by controlling a variety of signaling pathways and acting on a variety of targets.This has also been supported by in vitro cell studies,which serve as benchmarks for further research.
文摘The functional formula diet AHP, containing polysaccharides from Auricularia auricular, polyphenolic compounds from Hawthorn (Crataegus) and Pueraria radix, has been recently developed as a dietary intervention against dyslipidemia in our previous study. In the present study, its antioxidant activities and protective effects against proliferation of vascular smooth muscle cells (VSMCs) were investigated. AHP possessed the potent radical-scavenging effects against hydroxyl and superoxide radicals, and also the inhibitory effects against peroxidation of low density lipoprotein induced by Cu2+ in vitro. The protective effects of AHP against proliferation of VSMCs were evaluated through the methodology of serum pharmacology. The serum containing AHP significantly inhibited the proliferation of VSMCs induced by oxidized low density lipoprotein in a time- and dose-dependent manner, and also promoted the nitric oxide production of VSMCs. Our study indicated that this functional formula diet would be a potent alternative as a functional diet to prevent atherosclerosis at early stage.
基金Supported by Natural Science Research Project of Guangxi University of Chinese Medicine in 2015"Study on Anti-tumor Effect of Phyllanthus reticulatus Leaf Extract"(p15028)Independent Research Program of Guangxi Key Laboratory of Traditional Chinese Medicine Pharmacology in 2015"Study on Phyllanthus reticulatus Leaf 7404 Nude Mice Xenografts"+1 种基金Young Teachers Enhancement Project of Guangxi Department of Education in 2018"Study on the Mechanism of Zhuang Medicine Phyllanthus reticulatus Leaf against Liver Cancer"(2018KY0300)2019-2021 Guangxi First-class Discipline Construction Open Project Fund for Young Scholars of Guangxi University of Chinese Medicine"Anti-hepatocarcinoma Mechanism of Autophagy Induced by Ethyl Acetate in Phyllanthus reticulatus Leaf Based on PI3K/AKT and STAT3/m TOR Signaling Pathway"(2019XK089)
文摘[Objectives]To observe the effect of drug-containing serum from different extracts of Phyllanthus reticulatus leaf on the proliferation of human gastric cancer SGC-7901 cells and the effect of in vivo on the life extension rate of mice with H22 ascites tumor,and to investigate the effects of chemical components such as Corilagin,ethyl gallate,ethyl brevifolincarboxylate,and gallic acid on the proliferation of human gastric cancer SGC-7901 cells.[Methods]MTT method was used to observe the inhibitory effect of drug-containing serum of different extracts of P.reticulatus leaf on human gastric cancer SGC-7901 cells in vitro to determine its active site.The active site was used as the research object to establish a Kunming mouse ascites tumor model,to investigate the effect on the life extension rate of mice with H22 ascites tumor,to further separate the monomer components from the effective fraction and investigate its effect on human gastric cancer SGC-7901 cells.[Results]Compared with the 10%blank serum control group,10%drug-containing serum of ethyl acetate and n-butanol in P.reticulatus leaf had significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and the inhibition rate was 34.99%and 28.68%,respectively(P<0.05).Compared with the blank group,the survival time of the high dose group of ethyl acetate in P.reticulatus leaf was significant(P<0.05).Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester had a significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,with IC 50 of 26.52,70.45,and 158.86μg/mL,respectively.Ethyl gallate had no inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and its IC 50 was 251.96μg/mL.[Conclusions]The drug-containing serum of ethyl acetate and n-butanol extract of P.reticulatus can inhibit the proliferation of human gastric cancer SGC-7901 cells.Ethyl acetate of P.reticulatus leaves can increase the life extension rate of mice with H22 ascites tumor.Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester isolated from its active site are the material basis for inhibiting the proliferation of human gastric cancer SGC-7901 cells.
基金Supported by the Natural Science Foundation of China(No.30572438)
文摘Objective： To investigate the effect of Jinmaitong （筋脉通,JMT） serum on the proliferation of rat Schwann cells （SCs） primarily cultured in high glucose medium. Method： SOs were primarily cultured in Dulbecco＇s minmum essential medium （DMEM control）, 50 mmol/L glucose medium （50 mmol/L Glu）, 75 mmol/L glucose medium （75 mmol/L Glu）, as well as 50 mmol/L glucose medium, with different concentrations of JMT serum （undiluted, 1：2 diluted and 1：8 diluted） and Neurotropin （Ntp）, respectively. The proliferation of SCs under different conditions was detected by MTT. Result： SCs grew exuberantly in DMEM within 24-72 h, but slowed down at 96 h. The proliferation of SCs was inhibited in 50 mmol/L Glu and 75 mmol/L Glu after cultures of 48, 72 and 96 h, which showed that both were significantly different compared to the control group （P〈0.01）. The inhibition was more significant in 75 mmol/L Glu than in 50 mmol/L Glu （P〈0.05）. Spearman＇s rho analysis revealed that the proliferation of SCs had a negative correlation with the concentration of glucose （r=-0.471, P〈0.01）. Excluding the time factor, partial correlation showed similar results （r =-0.679, P〈0.01）. After 48 h, the proliferation of SCs increased significantly in JMT 1：2 and Ntp compared with 50 mmol/L Glu （control 0.437±0.019, 50 mmol/ L Glu 0.367±0.035, JMT1：2 0.426±0.024, Ntp 0.422±0.013; P〈0.01）, and there were no statistically significant differences among the JMT groups, the Ntp group and the control group （P〉0.05）. Conclusions： The proliferation of SCs was inhibited in high glucose medium, and the inhibition was reduced by different concentrations of JMT serum, especially at JMT1：2.
基金the National Natural Science Foundation of China，the Key Doctoral Foundation of Chinese Education Commission，the Beijing Natural Science Foundation
文摘Background The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood （PB-MSCs） were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair. Methods MSCs were mobilized into peripheral blood by granulocyte colony stimulating factor （G-CSF） and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension （20% 02 and 2% O2）, with or without 10% fetal bovine serum （FBS） conditions： 20% 02 and 10% FBS complete medium （normal medium, N）, 20% 02 and serum deprivation medium （D）, 2% 02 and 10% FBS complete medium （hypoxia, H）, 2% 02 and serum deprivation （HD）. Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling （TUNEL） staining. Results Spindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow （BM-MSCs）, and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II （MHC II）. They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia. Conclusions PB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.
