Objectives:This study aimed to reveal the role and possible mechanism of the ubiquitin-conjugating enzyme 2T(UBE2T)in the biological activities of breast cancer stem cells(BCSCs).Methods:The specific protein and gene ...Objectives:This study aimed to reveal the role and possible mechanism of the ubiquitin-conjugating enzyme 2T(UBE2T)in the biological activities of breast cancer stem cells(BCSCs).Methods:The specific protein and gene expression were quantified by Western blotting and quantitative real-time polymerase chain reaction,the proportion of BCSCs was examined by flow cytometry,and the self-renewal and proliferation of BCSCs were verified by serial sphere formation and soft agar.Results:Increasing expression of UBE2T was drastically found in breast cancer than that in adjacent tissues.Furthermore,UBE2T overexpression significantly increased the proportion of BCSCs in breast cancer cells and promoted their self-renewal and proliferation.Silent UBE2T exhibited the opposite functions.UBE2T increased the levels of the mammalian target of rapamycin and the phosphorylated mammalian target of rapamycin.Mammalian target of rapamycin(mTOR)inhibitor rapamycin inhibited the function of UBE2T in BCSCs.Conclusion:UBE2T plays a role in BCSCs through mTOR pathway and may suggest a novel therapeutic strategy for breast cancer.展开更多
Background: The absence of a safe and effective therapy for recurrent spontaneous abortion due to a maternofetal failure in immunological tolerance remains an intractable clinical obstacle for surgeons. Recently, trad...Background: The absence of a safe and effective therapy for recurrent spontaneous abortion due to a maternofetal failure in immunological tolerance remains an intractable clinical obstacle for surgeons. Recently, traditional Chinese medicine has become a feasible alternative for certain diseases, including recurrent spontaneous abortion. However, because of the complex composition of the traditional Chinese medicine formula, its action mechanism remains unclear. Methods: We selected two isolated active ingredients (RAMP and baicalin) from the traditional Chinese medicine formula and used an abortion-prone CBA/J × DBA/2 model to simulate human RSA and compared the changes in fetal resorption rate, Treg cell percentage, and relevant cytokines before and after combination therapy. In addition, The mechanisms were preliminarily discussed using in vitro differentiation models. Results: In CBA/J × DBA/2 abortion-prone mice, the combination therapy resulted in a lower embryo resorption rate compared to that obtained with individual delivery of either RAMP or baicalin, thereby playing an embryo-protective role through the increase in Treg cells for the maintenance of maternal-fetal immune tolerance. In in vitro primary cell differentiation experiments, the concentration of Treg cells significantly increased from 11% to 17.9% after the combination therapy compared to that of the single administration group. Conclusion: the synergistic effects of RAMP and baicalin were responsible for Treg differentiation. The present study provides a solid basis for improving the applicability of traditional Chinese herbs in the treatment of recurrent spontaneous abortion.展开更多
Objective:To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline(TG)for reducing the H_2O_2induced toxicity in H9c2 cells.Methods:Cyto...Objective:To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline(TG)for reducing the H_2O_2induced toxicity in H9c2 cells.Methods:Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit.RNA extraction and cDNA synthesized according to the kit manufacture protocol.Apoptosis was measured by the Flowcytometry,general PCR and qPCR.Results:It was found that the TG significantly rescued the morphology of the H9c2 cells.Treatment of cells with TG attenuated H_2O_2 induced cell deaths and improved the antioxidant activity.In addition,TG regulated the apoptotic gene caspase-3,caspase-9 and anti-apoptotic gene Bcl-2.Bcl-XL during H_2O_2 induced oxidative stress in H9c2 cells.These results were comparable with quercetin treatment.For evident,flow cytometer results also confirmed the TG significantly reduced the H_2O_2 induced necrosis and apoptosis in H9c2 cells.However,further increment of TG concentration against H_2O_2 could induce the necrosis and apoptosis along with H_2O_2.Conclusions:It is suggested that less than 125μM of TG could protect the cells from H_2O_2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression.Therefore,we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.展开更多
Pathological cardiac hypertrophy induced by angiotensin Ⅱ (Ang Ⅱ ) can subsequently give rise to heart failure, a leading cause of mortality. Nardosinone is a pharmacologically active compound extracted from the r...Pathological cardiac hypertrophy induced by angiotensin Ⅱ (Ang Ⅱ ) can subsequently give rise to heart failure, a leading cause of mortality. Nardosinone is a pharmacologically active compound extracted from the roots ofNardostachys chinensis, a well-known traditional Chinese medicine. In order to investigate the effects of nardosinone on Ang Ⅱ-induced cardiac cell hypertrophy and the related mechanisms, the myoblast cell line H9c2, derived from embryonic rat heart, was treated with nardosi- none (25, 50, 100, and 200μmol/L) or Ang Ⅱ (1 μmol/L). Then cell surface area and mRNA expression of classical markers of hypertrophy were detected. The related protein levels in PI3K/Akt/mTOR and MEK/ERK signaling pathways were examined by Western blotting. It was found that pretreatment with nardosinone could significantly inhibit the enlargement of cell surface area induced by Ang Ⅱ. The mRNA expression of ANP, BNP and 13-MHC was obviously elevated in Ang Ⅱ-treated H9c2 cells, which could be effectively blocked by nardosinone at the concentration of 100μmol/L. Further study revealed that the protective effects of nardosinone might be mediated by repressing the phosphorylation of related proteins in PI3K/Akt and MEK/ERK signaling pathways. It was suggested that the inhibitory effect of nardosinone on Ang Ⅱ-induced hypertrophy in H9c2 cells might be mediated by targeting PI3K/Akt and MEK/ERK signaling pathways.展开更多
This work aimed to study the beneficial role of human umbilical cord blood-derived mesenchymal stem cellconditioned medium(MSC-CM)in hypoxia-induced apoptosis in H9c2 cardiomyoblasts,in which the serine/heroine kinase...This work aimed to study the beneficial role of human umbilical cord blood-derived mesenchymal stem cellconditioned medium(MSC-CM)in hypoxia-induced apoptosis in H9c2 cardiomyoblasts,in which the serine/heroine kinases(Akt)pathway would be involved.For this,CM was collected by culturing MSCs in serum-free DMEM medium for 24 h,and paracrine factors were analyzed by protein chip.H9c2 cells were divided into the following groups:control group,hypoxia group,MSC-CM intervention group(CM group),MSC-CM+Akt phosphorylation inhibitor(LY294002)group(LY group).Apoptosis of the H9c2 cells was tested with chromatin dye Hoechst 33342 and FITC-conjugated Annexin V apoptosis detection kit by flow cytometer after a hypoxia/serum deprivation(H/SD)for 24 h.The apoptosis-related proteins were evaluated by Western blot.MSC-CM displayed significantly elevated levels of growth factors,anti-inflammatory,and anti-apoptosis cytokines.On Hoechst 33342 apoptosis staining,the H9c2 cell morphology displayed a lower proportion of apoptosis in the CM group than those in the hypoxia group,while apoptosis was increased in LY group.Flow cytometer analysis revealed the apoptosis ratio in the CM group was lower than the hypoxia group(12.34±2.00%vs.21.73±2.58%,p<0.05),while the LY group was significantly higher(22.54±3.89%).Active caspase-3 expression was increased in hypoxia group than control group(p<0.05),but decreased in CM group(p<0.01).Umbilical cord blood-derived mesenchymal stem cell-conditioned media secrete multiple paracrine factors that are able to inhibit hypoxia-induced H9c2 cardiomyoblasts apoptosis,and in which the activation of Akt phosphorylation is involved to achieve the protective effect.展开更多
This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T(HEK293T) cells induced by cadmium chloride(CdCl2) and nanometer titanium dioxide(nano-TiO2).RT-PCR technique was used t...This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T(HEK293T) cells induced by cadmium chloride(CdCl2) and nanometer titanium dioxide(nano-TiO2).RT-PCR technique was used to detect the expressions of Heme oxygenase-1(HO-1) and 8-oxoguanine DNA glycosylase(OGG1).The activities of superoxide dismutase(SOD) and catalase enzyme(CAT) and concentrations of reactive oxygen species(ROS) and maldondialdehyde(MDA) were measured by different approaches.The results showed that CdCl2 and nano-TiO2 at a low concen-tration of 0.75 total toxic unit(TU) exerted an additive effects on HO-1 gene expression,CAT activities and MDA concentrations.When the total TU was increased to 1 or 1.25 TU,the interaction was syner-getic.Moreover,the mixture with high proportion of CdCl2 produced an additive effect on the OGG1 gene expression,and the interaction was changed to be synergetic when the concentration of CdCl2 was lower than or equal to that of nano-TiO2.Synergetic effects of CdCl2 and nano-TiO2 on cellular oxida-tive damage of HEK293T cells were found as indicated by the changes in the SOD activities and ROS concentrations.It was concluded that CdCl2 and nano-TiO2 exerts synergistic effects on the cellular oxidative damage of HEK293T cells,and the sensitivity of these indicators of oxidative damage varies with the proportion of CdCl2 and nano-TiO2 in the mixture.展开更多
OBJECTIVE We aimed identification of cell surface molecules, which might serve as diagnostic biomarkers or useful targets for therapies, in breast cancer. METHODS We developed unique DNA microarray coupled with spheri...OBJECTIVE We aimed identification of cell surface molecules, which might serve as diagnostic biomarkers or useful targets for therapies, in breast cancer. METHODS We developed unique DNA microarray coupled with spherical self-organizing map (sSOM) analysis to characterize cells and tissues by the cell surface markers. In the microarray 1,797 probes for human genes coding membrane bound proteins were spotted. With this microarray the gene expression profiles of eight breast carcinoma cell lines were compared to identify the genes that were commonly expressed in breast carcinomas but not in normal cells. RESULTS The gene expression profiles of sSOM from the eight breast carcinoma cell lines were successfully distinguished from that of normal breast tissue derived cells suggesting the presence of genes of interest, sSOMon the data extensively filtered revealed several candidate genes, of which expression was significant in carcinoma cells but low in normal cells. Finally, TM9SF2 was nominated through validations of PCR procedures together with CD24 and ErbB3, which are known breast carcinoma markers. TMgSF2 expression was further confirmed by immunological staining. Interestingly, TMgSF2 was found to be expressed in all the cell lines evaluated while CD24 and ErbB3 were not in all of the carcinoma cells, supporting their relationship in sSOM. Although physiological significance of TMgSF2 is unknown yet, siRNA treatment significantly inhibited the growth of MDA- MB-231 cells. CONCLUSION We propose TM9SF2 as a novel and useful diagnostic marker as well as a potential molecular target specific to breast carcinoma cells covering wide range of breast cancer.展开更多
OBJECTIVE Liguzinediol is a derivative of the natural active ingredient ligustrazine,and we found that liguzinediol has significant positive inotropic effects,which are stronger than that of TMP.Besides,it does not le...OBJECTIVE Liguzinediol is a derivative of the natural active ingredient ligustrazine,and we found that liguzinediol has significant positive inotropic effects,which are stronger than that of TMP.Besides,it does not lead to arrhythmia,hypotension and other side effects.This study aims to investigate the anti-apoptotic effects of liguzinediolon H9C2 cells.METHODS Apoptotic H9C2 cells induced by DOX were observed by electron microscope and FCM analysis.The protein expressions of Bax,Bcl-2,caspases 3 and NF-κB were detected by Western blotting.RESULTS Apoptotic H9C2 cells induced by DOX were observed,but without apoptotic bodies in liguzinediol group.Declined peak of H9C2 cell apoptosis was seen in liguzinediol group by FCM analysis.And downregulation of Bax,caspases 3,NF-κB and upregulation of Bcl-2 were found by Western blotting.CONCLUSION Liguzinediol protected cardiomyocytes against apoptosis through downregulation of Bax and caspases 3 and upregulation of Bcl-2.Liguzinediol can inhibited cardiomyocyte apoptosis through the NF-κB signal pathway.展开更多
Cilia depend on their highly differentiated structure, a 9 + 2 arrangement, to remove particles from the lung and to transport reproductive cells. Immortalized cells could potentially be of great use in cilia researc...Cilia depend on their highly differentiated structure, a 9 + 2 arrangement, to remove particles from the lung and to transport reproductive cells. Immortalized cells could potentially be of great use in cilia research. Immortalization of cells with cilia structure containing the 9 + 2 arrangement might be able to generate cell lines with such cilia structure. How- ever, whether immortalized cells can retain such a highly differentiated structure remains unclear. Here we demonstrate that (1) using Ela gene transfection, tracheal cells are immortalized; (2) interestingly, in a gel culture the immortalized cells form spherical aggregations within which a lumen is developed; and (3) surprisingly, inside the aggregation, cilia containing a 9 + 2 arrangement grow from the cell's apical pole and protrude into the lumen. These results may influence future research in many areas such as understanding the mechanisms of cilia differentiation, cilia generation in other existing cell lines, cilia disorders, generation of other highly differentiated structures besides cilia using the gel culture, immortalization of other ciliated cells with the Ela gene, development of cilia motile function, and establishment of a research model to provide uniform ciliated cells.展开更多
OBJECTIVE Aconitine(ACO)as the main active component in Aconitum carmichaelii debeaux(family Ranunlaceae),has highly toxicity in heart and the mechanisms are not clear yet.Paeoniflorin(PF),the main chemical ingredient...OBJECTIVE Aconitine(ACO)as the main active component in Aconitum carmichaelii debeaux(family Ranunlaceae),has highly toxicity in heart and the mechanisms are not clear yet.Paeoniflorin(PF),the main chemical ingredient in Herbaceous peony,can protect heart hurt by antioxidant,vasodilator effect and other effects.In this study,we focused on investigating the mechanism of PF reducing the cardiotoxicity of ACO.METHODS We chose H9c2 cells as experimental subject.MTT,Western blotting and real-time PCR were used to measure cell proliferation,apoptosis,ion channels and oxidative stress.RESULTS Cell proliferation in ACO+PF group was significantly increased compared with ACO group;the ratio with Bcl-2 and Bax and the level of p53 were upregulated by PF,while the level of caspase-3 was lightly reduced.The expression of SCN5A mRNA significantly was increased in ACO+PF group,while the expres⁃sion of RyR2 and Cx43 mRNA was dropped.Compared with ACO group,extracellular LDH and intracellular MDA were highly decreased,while intracellular SOD was regulated.CONCLUSION Cardiotoxicity of ACO in H9c2 cells was signifi⁃cantly decreased by PF.展开更多
The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative di...The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative diseases. In the present study, we observed the effects of transfecting three amyloid precursor-like protein 2 (APLP2) C-terminal fragments (CTFs; C57, C50 and C31) in the pEGFP-N1 vector on S100A9 expression in BV2 cells. Reverse transcription-PCR, western blot assay and immunocytochemistry revealed that S100A9 protein and mRNA expression was greater in BV2 cells after CTF transfection than after mock transfection with an empty vector. Furthermore, transfection of full-length APLP2-751 resulted in low levels of S100A9 protein ex-pression. Our results show that APLP2-CTFs upregulate S100A9 protein and mRNA expression in BV2 cells, and identify a novel pathway involved in neuronal injury and apoptosis, and repair and protection in Alzheimer’s disease.展开更多
Daji (Cirsium japonicum) has been applied against gastric disorders, lung diseases, and cardiovascular problems in thetraditional Chinese medicinal system. The present study was to investigate the protective effects...Daji (Cirsium japonicum) has been applied against gastric disorders, lung diseases, and cardiovascular problems in thetraditional Chinese medicinal system. The present study was to investigate the protective effects of Daji (Cirsiumjaponicum) polysaccharide extracts (CJP) against hydrogen peroxide (H2O2) shock in rat H9c2 myocardial cells. First,CJP was isolated by hot water extraction and ethanol precipitation; it was then characterized by high performance liquidchromatography and infrared spectrum analysis. Rat H9c2 cells were subjected to H2O2 treatment to establish a cellinjury model. The 3- (4,5- dimethylthiazol- 2-yl)-2,5- diphenyltetrazolium bromide assay showed that CJP pretreatmentsignificantly ameliorated the H2O2 injury in a dose-dependent manner. Furthermore, the cell apoptosis induced by H2O2was markedly inhibited by CJP pretreatment, whereas the cleavage level of caspase-3, -8, and -9 was reduced. Inaddition, the p38 mitogen-activated protein kinase pathway might be involved in the protective effect of CJP onmyocardial cells. Therefore, we conclude that polysaccharide extracts of Daji (Cirsium japonicum) protect rat H9c2myocardial cells from oxidative stress induced by H2O2.展开更多
[Objective] The study aims to determine the optimal concentration of trypsin for the proliferation of avian influenza virus (AIV) H9N2 subtype in Madin- Darby canine kidney (MDCK) cells. [Method] Three AIV H9 subt...[Objective] The study aims to determine the optimal concentration of trypsin for the proliferation of avian influenza virus (AIV) H9N2 subtype in Madin- Darby canine kidney (MDCK) cells. [Method] Three AIV H9 subtype isolates were inoculated on MDCK cells respectively. Then, DMEM containing different concentrations of trypsin as maintenance media were added to MDCK monolayer cells. The cytopathic effect (CPE) was observed once every 24 h, and the HA titer of the supematant was measured by HA assay. [Result] When the trypsin concentration was 10 -20 μg/ml in DMEM, the HA titer of virus culture reached 7 log2 (1:128). Almost all cells were cytopathic after 96 h post inoculation with 1:1 000 or 1:10 000 dilution of AIV culture, and the virus titer reached a peak after 72 -96 h. [ Conclusion] The optimal concentration of trypsin is 10 -20 pg/ml for proliferation of AIV H9N2 subtype in MDCK cells.展开更多
Objective: To study the effect of different doses of doxorubicin on H9C2 cells and to provide a reference for the clinical study of doxorubicin. Methods: Doxorubicin (1, 2, 4, 6, 10 ug/ml) was co-cultured with H9C2 ce...Objective: To study the effect of different doses of doxorubicin on H9C2 cells and to provide a reference for the clinical study of doxorubicin. Methods: Doxorubicin (1, 2, 4, 6, 10 ug/ml) was co-cultured with H9C2 cells for 6, 12 and 24 hours. The morphological changes of cells were observed, and the cell inhibition rates of different time and drug concentration were calculated. Results: Doxorubicin could inhibit the activity of cardiomyocytes in a dose-dependent manner from 1 to 10 ug/ml. Conclusion: A certain dose of doxorubicin has a toxic effect on cardiomyocytes and can cause cardiomyocyte necrosis and apoptosis.展开更多
Developing reliable and efficient anode materials is essential for the successfully practical application of sodium-ion batteries.Herein,employing a straightforward and rapid chemical vapor deposition technique,two-di...Developing reliable and efficient anode materials is essential for the successfully practical application of sodium-ion batteries.Herein,employing a straightforward and rapid chemical vapor deposition technique,two-dimensional layered ternary indium phosphorus sulfide(In_(2)P_(3)S_(9)) nanosheets are prepared.The layered structure and ternary composition of the In_(2)P_(3)S_(9) electrode result in impressive electrochemical performance,including a high reversible capacity of 704 mA h g^(-1) at 0.1 A g^(-1),an outstanding rate capability with 425 mA h g^(-1) at 5 A g^(-1),and an exceptional cycling stability with a capacity retention of88% after 350 cycles at 1 A g^(-1).Furthermore,sodium-ion full cell also affords a high capacity of 308 and114 mA h g^(-1) at 0.1 and 5 A g^(-1).Ex-situ X-ray diffraction and ex-situ high-resolution transmission electron microscopy tests are conducted to investigate the underlying Na-storage mechanism of In_(2)P_(3)S_(9).The results reveal that during the first cycle,the P-S bond is broken to form the elemental P and In_(2)S_(3),collectively contributing to a remarkably high reversible specific capacity.The excellent electrochemical energy storage results corroborate the practical application potential of In_(2)P_(3)S_(9) for sodium-ion batteries.展开更多
基金This research was partly supported by the Fundamental Research Funds of Shandong University(21510078614097)the Shandong Natural Science Foundation General Project(ZR2022MC093).
文摘Objectives:This study aimed to reveal the role and possible mechanism of the ubiquitin-conjugating enzyme 2T(UBE2T)in the biological activities of breast cancer stem cells(BCSCs).Methods:The specific protein and gene expression were quantified by Western blotting and quantitative real-time polymerase chain reaction,the proportion of BCSCs was examined by flow cytometry,and the self-renewal and proliferation of BCSCs were verified by serial sphere formation and soft agar.Results:Increasing expression of UBE2T was drastically found in breast cancer than that in adjacent tissues.Furthermore,UBE2T overexpression significantly increased the proportion of BCSCs in breast cancer cells and promoted their self-renewal and proliferation.Silent UBE2T exhibited the opposite functions.UBE2T increased the levels of the mammalian target of rapamycin and the phosphorylated mammalian target of rapamycin.Mammalian target of rapamycin(mTOR)inhibitor rapamycin inhibited the function of UBE2T in BCSCs.Conclusion:UBE2T plays a role in BCSCs through mTOR pathway and may suggest a novel therapeutic strategy for breast cancer.
基金the National Natural Science Foundation of China(81973221)National Natural Science Foundation for Young Scientists of China(81603647)+2 种基金the Women and Children Health Talent Project of Jiangsu Province(FRC201785)the Chinese Clinical Medicine Innovation Center of Obstetrics,Gynecology,and Reproduction in Jiangsu Province(ZX202102)the Women and Children Health Science Foundation of Jiangsu Province(F202206).
文摘Background: The absence of a safe and effective therapy for recurrent spontaneous abortion due to a maternofetal failure in immunological tolerance remains an intractable clinical obstacle for surgeons. Recently, traditional Chinese medicine has become a feasible alternative for certain diseases, including recurrent spontaneous abortion. However, because of the complex composition of the traditional Chinese medicine formula, its action mechanism remains unclear. Methods: We selected two isolated active ingredients (RAMP and baicalin) from the traditional Chinese medicine formula and used an abortion-prone CBA/J × DBA/2 model to simulate human RSA and compared the changes in fetal resorption rate, Treg cell percentage, and relevant cytokines before and after combination therapy. In addition, The mechanisms were preliminarily discussed using in vitro differentiation models. Results: In CBA/J × DBA/2 abortion-prone mice, the combination therapy resulted in a lower embryo resorption rate compared to that obtained with individual delivery of either RAMP or baicalin, thereby playing an embryo-protective role through the increase in Treg cells for the maintenance of maternal-fetal immune tolerance. In in vitro primary cell differentiation experiments, the concentration of Treg cells significantly increased from 11% to 17.9% after the combination therapy compared to that of the single administration group. Conclusion: the synergistic effects of RAMP and baicalin were responsible for Treg differentiation. The present study provides a solid basis for improving the applicability of traditional Chinese herbs in the treatment of recurrent spontaneous abortion.
文摘Objective:To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline(TG)for reducing the H_2O_2induced toxicity in H9c2 cells.Methods:Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit.RNA extraction and cDNA synthesized according to the kit manufacture protocol.Apoptosis was measured by the Flowcytometry,general PCR and qPCR.Results:It was found that the TG significantly rescued the morphology of the H9c2 cells.Treatment of cells with TG attenuated H_2O_2 induced cell deaths and improved the antioxidant activity.In addition,TG regulated the apoptotic gene caspase-3,caspase-9 and anti-apoptotic gene Bcl-2.Bcl-XL during H_2O_2 induced oxidative stress in H9c2 cells.These results were comparable with quercetin treatment.For evident,flow cytometer results also confirmed the TG significantly reduced the H_2O_2 induced necrosis and apoptosis in H9c2 cells.However,further increment of TG concentration against H_2O_2 could induce the necrosis and apoptosis along with H_2O_2.Conclusions:It is suggested that less than 125μM of TG could protect the cells from H_2O_2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression.Therefore,we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.
基金supported by the grants from the National Natural Science Foundation of China(No.30971245 and No.81000112)
文摘Pathological cardiac hypertrophy induced by angiotensin Ⅱ (Ang Ⅱ ) can subsequently give rise to heart failure, a leading cause of mortality. Nardosinone is a pharmacologically active compound extracted from the roots ofNardostachys chinensis, a well-known traditional Chinese medicine. In order to investigate the effects of nardosinone on Ang Ⅱ-induced cardiac cell hypertrophy and the related mechanisms, the myoblast cell line H9c2, derived from embryonic rat heart, was treated with nardosi- none (25, 50, 100, and 200μmol/L) or Ang Ⅱ (1 μmol/L). Then cell surface area and mRNA expression of classical markers of hypertrophy were detected. The related protein levels in PI3K/Akt/mTOR and MEK/ERK signaling pathways were examined by Western blotting. It was found that pretreatment with nardosinone could significantly inhibit the enlargement of cell surface area induced by Ang Ⅱ. The mRNA expression of ANP, BNP and 13-MHC was obviously elevated in Ang Ⅱ-treated H9c2 cells, which could be effectively blocked by nardosinone at the concentration of 100μmol/L. Further study revealed that the protective effects of nardosinone might be mediated by repressing the phosphorylation of related proteins in PI3K/Akt and MEK/ERK signaling pathways. It was suggested that the inhibitory effect of nardosinone on Ang Ⅱ-induced hypertrophy in H9c2 cells might be mediated by targeting PI3K/Akt and MEK/ERK signaling pathways.
基金the National Natural Science Fund of China(81600196)grants from Beijing key laboratory of metabolic disorder related cardiovascular disease.
文摘This work aimed to study the beneficial role of human umbilical cord blood-derived mesenchymal stem cellconditioned medium(MSC-CM)in hypoxia-induced apoptosis in H9c2 cardiomyoblasts,in which the serine/heroine kinases(Akt)pathway would be involved.For this,CM was collected by culturing MSCs in serum-free DMEM medium for 24 h,and paracrine factors were analyzed by protein chip.H9c2 cells were divided into the following groups:control group,hypoxia group,MSC-CM intervention group(CM group),MSC-CM+Akt phosphorylation inhibitor(LY294002)group(LY group).Apoptosis of the H9c2 cells was tested with chromatin dye Hoechst 33342 and FITC-conjugated Annexin V apoptosis detection kit by flow cytometer after a hypoxia/serum deprivation(H/SD)for 24 h.The apoptosis-related proteins were evaluated by Western blot.MSC-CM displayed significantly elevated levels of growth factors,anti-inflammatory,and anti-apoptosis cytokines.On Hoechst 33342 apoptosis staining,the H9c2 cell morphology displayed a lower proportion of apoptosis in the CM group than those in the hypoxia group,while apoptosis was increased in LY group.Flow cytometer analysis revealed the apoptosis ratio in the CM group was lower than the hypoxia group(12.34±2.00%vs.21.73±2.58%,p<0.05),while the LY group was significantly higher(22.54±3.89%).Active caspase-3 expression was increased in hypoxia group than control group(p<0.05),but decreased in CM group(p<0.01).Umbilical cord blood-derived mesenchymal stem cell-conditioned media secrete multiple paracrine factors that are able to inhibit hypoxia-induced H9c2 cardiomyoblasts apoptosis,and in which the activation of Akt phosphorylation is involved to achieve the protective effect.
基金the National Investigation Projects of Soil Pollution supported by a grant from Ministry of Environmental Protection of the People's Republic of China
文摘This study investigated the conjoined cellular oxidative damage of human embryo kidney 293T(HEK293T) cells induced by cadmium chloride(CdCl2) and nanometer titanium dioxide(nano-TiO2).RT-PCR technique was used to detect the expressions of Heme oxygenase-1(HO-1) and 8-oxoguanine DNA glycosylase(OGG1).The activities of superoxide dismutase(SOD) and catalase enzyme(CAT) and concentrations of reactive oxygen species(ROS) and maldondialdehyde(MDA) were measured by different approaches.The results showed that CdCl2 and nano-TiO2 at a low concen-tration of 0.75 total toxic unit(TU) exerted an additive effects on HO-1 gene expression,CAT activities and MDA concentrations.When the total TU was increased to 1 or 1.25 TU,the interaction was syner-getic.Moreover,the mixture with high proportion of CdCl2 produced an additive effect on the OGG1 gene expression,and the interaction was changed to be synergetic when the concentration of CdCl2 was lower than or equal to that of nano-TiO2.Synergetic effects of CdCl2 and nano-TiO2 on cellular oxida-tive damage of HEK293T cells were found as indicated by the changes in the SOD activities and ROS concentrations.It was concluded that CdCl2 and nano-TiO2 exerts synergistic effects on the cellular oxidative damage of HEK293T cells,and the sensitivity of these indicators of oxidative damage varies with the proportion of CdCl2 and nano-TiO2 in the mixture.
基金supported by the Grantin-Aid for scientific research(B)No.18300164"Screening of carcinoma cell surface markers and its application in molecular targeting with bionanocapsules"Japan Society for the Promotion of Science(JSPS).
文摘OBJECTIVE We aimed identification of cell surface molecules, which might serve as diagnostic biomarkers or useful targets for therapies, in breast cancer. METHODS We developed unique DNA microarray coupled with spherical self-organizing map (sSOM) analysis to characterize cells and tissues by the cell surface markers. In the microarray 1,797 probes for human genes coding membrane bound proteins were spotted. With this microarray the gene expression profiles of eight breast carcinoma cell lines were compared to identify the genes that were commonly expressed in breast carcinomas but not in normal cells. RESULTS The gene expression profiles of sSOM from the eight breast carcinoma cell lines were successfully distinguished from that of normal breast tissue derived cells suggesting the presence of genes of interest, sSOMon the data extensively filtered revealed several candidate genes, of which expression was significant in carcinoma cells but low in normal cells. Finally, TM9SF2 was nominated through validations of PCR procedures together with CD24 and ErbB3, which are known breast carcinoma markers. TMgSF2 expression was further confirmed by immunological staining. Interestingly, TMgSF2 was found to be expressed in all the cell lines evaluated while CD24 and ErbB3 were not in all of the carcinoma cells, supporting their relationship in sSOM. Although physiological significance of TMgSF2 is unknown yet, siRNA treatment significantly inhibited the growth of MDA- MB-231 cells. CONCLUSION We propose TM9SF2 as a novel and useful diagnostic marker as well as a potential molecular target specific to breast carcinoma cells covering wide range of breast cancer.
基金The project supported by the National Natural Science Foundation of China(81072542)Natural Science Foundation of Jiangsu Province(BK2011077)the Research Fund for the Doctoral Program of Higher Education(20123237110010)
文摘OBJECTIVE Liguzinediol is a derivative of the natural active ingredient ligustrazine,and we found that liguzinediol has significant positive inotropic effects,which are stronger than that of TMP.Besides,it does not lead to arrhythmia,hypotension and other side effects.This study aims to investigate the anti-apoptotic effects of liguzinediolon H9C2 cells.METHODS Apoptotic H9C2 cells induced by DOX were observed by electron microscope and FCM analysis.The protein expressions of Bax,Bcl-2,caspases 3 and NF-κB were detected by Western blotting.RESULTS Apoptotic H9C2 cells induced by DOX were observed,but without apoptotic bodies in liguzinediol group.Declined peak of H9C2 cell apoptosis was seen in liguzinediol group by FCM analysis.And downregulation of Bax,caspases 3,NF-κB and upregulation of Bcl-2 were found by Western blotting.CONCLUSION Liguzinediol protected cardiomyocytes against apoptosis through downregulation of Bax and caspases 3 and upregulation of Bcl-2.Liguzinediol can inhibited cardiomyocyte apoptosis through the NF-κB signal pathway.
文摘Cilia depend on their highly differentiated structure, a 9 + 2 arrangement, to remove particles from the lung and to transport reproductive cells. Immortalized cells could potentially be of great use in cilia research. Immortalization of cells with cilia structure containing the 9 + 2 arrangement might be able to generate cell lines with such cilia structure. How- ever, whether immortalized cells can retain such a highly differentiated structure remains unclear. Here we demonstrate that (1) using Ela gene transfection, tracheal cells are immortalized; (2) interestingly, in a gel culture the immortalized cells form spherical aggregations within which a lumen is developed; and (3) surprisingly, inside the aggregation, cilia containing a 9 + 2 arrangement grow from the cell's apical pole and protrude into the lumen. These results may influence future research in many areas such as understanding the mechanisms of cilia differentiation, cilia generation in other existing cell lines, cilia disorders, generation of other highly differentiated structures besides cilia using the gel culture, immortalization of other ciliated cells with the Ela gene, development of cilia motile function, and establishment of a research model to provide uniform ciliated cells.
文摘OBJECTIVE Aconitine(ACO)as the main active component in Aconitum carmichaelii debeaux(family Ranunlaceae),has highly toxicity in heart and the mechanisms are not clear yet.Paeoniflorin(PF),the main chemical ingredient in Herbaceous peony,can protect heart hurt by antioxidant,vasodilator effect and other effects.In this study,we focused on investigating the mechanism of PF reducing the cardiotoxicity of ACO.METHODS We chose H9c2 cells as experimental subject.MTT,Western blotting and real-time PCR were used to measure cell proliferation,apoptosis,ion channels and oxidative stress.RESULTS Cell proliferation in ACO+PF group was significantly increased compared with ACO group;the ratio with Bcl-2 and Bax and the level of p53 were upregulated by PF,while the level of caspase-3 was lightly reduced.The expression of SCN5A mRNA significantly was increased in ACO+PF group,while the expres⁃sion of RyR2 and Cx43 mRNA was dropped.Compared with ACO group,extracellular LDH and intracellular MDA were highly decreased,while intracellular SOD was regulated.CONCLUSION Cardiotoxicity of ACO in H9c2 cells was signifi⁃cantly decreased by PF.
基金supported by the Natural Science Foundation of Technology Gallery in Jilin Province of China,No.2011-15237the National Natural Science Foundation of China,No.81160159
文摘The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative diseases. In the present study, we observed the effects of transfecting three amyloid precursor-like protein 2 (APLP2) C-terminal fragments (CTFs; C57, C50 and C31) in the pEGFP-N1 vector on S100A9 expression in BV2 cells. Reverse transcription-PCR, western blot assay and immunocytochemistry revealed that S100A9 protein and mRNA expression was greater in BV2 cells after CTF transfection than after mock transfection with an empty vector. Furthermore, transfection of full-length APLP2-751 resulted in low levels of S100A9 protein ex-pression. Our results show that APLP2-CTFs upregulate S100A9 protein and mRNA expression in BV2 cells, and identify a novel pathway involved in neuronal injury and apoptosis, and repair and protection in Alzheimer’s disease.
文摘Daji (Cirsium japonicum) has been applied against gastric disorders, lung diseases, and cardiovascular problems in thetraditional Chinese medicinal system. The present study was to investigate the protective effects of Daji (Cirsiumjaponicum) polysaccharide extracts (CJP) against hydrogen peroxide (H2O2) shock in rat H9c2 myocardial cells. First,CJP was isolated by hot water extraction and ethanol precipitation; it was then characterized by high performance liquidchromatography and infrared spectrum analysis. Rat H9c2 cells were subjected to H2O2 treatment to establish a cellinjury model. The 3- (4,5- dimethylthiazol- 2-yl)-2,5- diphenyltetrazolium bromide assay showed that CJP pretreatmentsignificantly ameliorated the H2O2 injury in a dose-dependent manner. Furthermore, the cell apoptosis induced by H2O2was markedly inhibited by CJP pretreatment, whereas the cleavage level of caspase-3, -8, and -9 was reduced. Inaddition, the p38 mitogen-activated protein kinase pathway might be involved in the protective effect of CJP onmyocardial cells. Therefore, we conclude that polysaccharide extracts of Daji (Cirsium japonicum) protect rat H9c2myocardial cells from oxidative stress induced by H2O2.
基金funded by the Beijing Academy of Agriculture and Forestry Sciences (2010A007)
文摘[Objective] The study aims to determine the optimal concentration of trypsin for the proliferation of avian influenza virus (AIV) H9N2 subtype in Madin- Darby canine kidney (MDCK) cells. [Method] Three AIV H9 subtype isolates were inoculated on MDCK cells respectively. Then, DMEM containing different concentrations of trypsin as maintenance media were added to MDCK monolayer cells. The cytopathic effect (CPE) was observed once every 24 h, and the HA titer of the supematant was measured by HA assay. [Result] When the trypsin concentration was 10 -20 μg/ml in DMEM, the HA titer of virus culture reached 7 log2 (1:128). Almost all cells were cytopathic after 96 h post inoculation with 1:1 000 or 1:10 000 dilution of AIV culture, and the virus titer reached a peak after 72 -96 h. [ Conclusion] The optimal concentration of trypsin is 10 -20 pg/ml for proliferation of AIV H9N2 subtype in MDCK cells.
文摘Objective: To study the effect of different doses of doxorubicin on H9C2 cells and to provide a reference for the clinical study of doxorubicin. Methods: Doxorubicin (1, 2, 4, 6, 10 ug/ml) was co-cultured with H9C2 cells for 6, 12 and 24 hours. The morphological changes of cells were observed, and the cell inhibition rates of different time and drug concentration were calculated. Results: Doxorubicin could inhibit the activity of cardiomyocytes in a dose-dependent manner from 1 to 10 ug/ml. Conclusion: A certain dose of doxorubicin has a toxic effect on cardiomyocytes and can cause cardiomyocyte necrosis and apoptosis.
基金Financial supports from the National Natural Science Foundation of China(22265018 and 21961019)the Key Project of Natural Science Foundation of Jiangxi Province(20232ACB204010)。
文摘Developing reliable and efficient anode materials is essential for the successfully practical application of sodium-ion batteries.Herein,employing a straightforward and rapid chemical vapor deposition technique,two-dimensional layered ternary indium phosphorus sulfide(In_(2)P_(3)S_(9)) nanosheets are prepared.The layered structure and ternary composition of the In_(2)P_(3)S_(9) electrode result in impressive electrochemical performance,including a high reversible capacity of 704 mA h g^(-1) at 0.1 A g^(-1),an outstanding rate capability with 425 mA h g^(-1) at 5 A g^(-1),and an exceptional cycling stability with a capacity retention of88% after 350 cycles at 1 A g^(-1).Furthermore,sodium-ion full cell also affords a high capacity of 308 and114 mA h g^(-1) at 0.1 and 5 A g^(-1).Ex-situ X-ray diffraction and ex-situ high-resolution transmission electron microscopy tests are conducted to investigate the underlying Na-storage mechanism of In_(2)P_(3)S_(9).The results reveal that during the first cycle,the P-S bond is broken to form the elemental P and In_(2)S_(3),collectively contributing to a remarkably high reversible specific capacity.The excellent electrochemical energy storage results corroborate the practical application potential of In_(2)P_(3)S_(9) for sodium-ion batteries.