葛根素抑制阿霉素诱导的H9c2心肌细胞凋亡的实验研究
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摘要
目的观察葛根素对阿霉素诱导的H9c2心肌细胞凋亡的影响,并探讨其潜在的分子机制。方法H9c2心肌细胞分为空白对照组、阿霉素诱导损伤组和葛根素低、中、高浓度处理组。除空白对照组不做任何处理外,其他各组加入阿霉素及相应浓度葛根素。采用MTT法检测细胞活性,流式细胞术检测细胞凋亡率,Western blot法检测各组Caspase-3、Cleaved-caspase-3、Akt和p-Akt蛋白表达水平的变化。结果与空白对照组比较,阿霉素诱导损伤组细胞凋亡率、Cleaved-caspase-3/Caspase-3比值增加,p-Akt/Akt比值降低,差异均具有统计学意义(P<0.05)。与阿霉素诱导损伤组比较,葛根素高浓度处理组细胞凋亡率、Cleaved-caspase-3/Caspase-3比值降低,p-Akt/Akt比值增加,差异均具有统计学意义(P<0.05)。结论葛根素可能通过激活Akt蛋白来抑制阿霉素诱导的H9c2心肌细胞凋亡,提示葛根素可能是一种潜在的心肌保护剂。
Objective To observe the effect of puerarin on adriamycin-induced apoptosis of H9 c2 cells and to explore its molecular mechanism. Methods H9 c2 cells were divided into control group,adriamycin-induced injury group and puerarin low,medium and high concentration treatment groups. Except for control group,other groups were administrated with adriamycin and with different concentrations of puerarin. The cell viability was detected by MTT assay; flowcytometry was used to assess the apoptosis rate. The expression of Caspase-3,Cleaved-caspase-3,AKT and p-Akt were evaluated by Western blot. Results Compared with blank group,the apoptosis rate,the proportion of Cleaved-caspase-3/Caspase-3 increased,and the proportion of p-Akt/Akt decreased in adriamycin-induced injury group(P<0.05). Compared with adriamycin-induced injury group,the apoptosis rate,the proportion of Cleaved-caspase-3/Caspase-3,and the proportion of p-Akt/Akt increased in high concentration puerarin group(P<0.05). Conclusion Puerarin may inhibit the apoptosis of H9 c2 cells induced by adriamycin by activating Akt signaling,suggesting that puerarin may be a potential cardioprotective agent for use in the clinical treatment of cardiomyopathy triggered by adriamycin.
Objective To observe the effect of puerarin on adriamycin-induced apoptosis of H9 c2 cells and to explore its molecular mechanism. Methods H9 c2 cells were divided into control group,adriamycin-induced injury group and puerarin low,medium and high concentration treatment groups. Except for control group,other groups were administrated with adriamycin and with different concentrations of puerarin. The cell viability was detected by MTT assay; flowcytometry was used to assess the apoptosis rate. The expression of Caspase-3,Cleaved-caspase-3,AKT and p-Akt were evaluated by Western blot. Results Compared with blank group,the apoptosis rate,the proportion of Cleaved-caspase-3/Caspase-3 increased,and the proportion of p-Akt/Akt decreased in adriamycin-induced injury group(P<0.05). Compared with adriamycin-induced injury group,the apoptosis rate,the proportion of Cleaved-caspase-3/Caspase-3,and the proportion of p-Akt/Akt increased in high concentration puerarin group(P<0.05). Conclusion Puerarin may inhibit the apoptosis of H9 c2 cells induced by adriamycin by activating Akt signaling,suggesting that puerarin may be a potential cardioprotective agent for use in the clinical treatment of cardiomyopathy triggered by adriamycin.
引文
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[14]Xing G,Dong M,Li X,et al.Neuroprotective effects of puerarin against beta-amyloid-induced neurotoxicity in PC12 cells via a PI3K-dependent signaling pathway[J].Brain Res Bull,2011,85(3-4):212-218.
[15]李聪,周捷,黄海潮,等.葛根素对表柔比星致H9c2心肌细胞凋亡的影响[J].中国药房,2016(34):4807-4810.
[2]Tap WD,Papai Z,Van Tine BA,et al.Doxorubicin plus evofosfamide versus doxorubicin alone in locally advanced,unresectable or metastatic soft-tissue sarcoma(TH CR-406/SARC021):an international,multicentre,open-label,randomised phase 3 trial[J].Lancet Oncol,2017,18(8):1089-1103.
[3]Olivieri J,Perna GP,Bocci C,et al.Modern Management of Anthracycline-Induced Cardiotoxicity in Lymphoma Patients:Low Occurrence of Cardiotoxicity with Comprehensive Assessment and Tailored Substitution by Nonpegylated Liposomal Doxorubicin[J].Oncologist,2017,22(4):422-431.
[4]Shi Y,Moon M,Dawood S,et al.Mechanisms and management of doxorubicin cardiotoxicity[J].Herz,2011,36(4):296-305.
[5]胥甜甜.葛根素抗心肌细胞缺氧复氧损伤保护作用机制研究[D].南昌:南昌大学,2014.
[6]Zhou YX,Zhang H,Peng C.Puerarin:a review of pharmacological effects[J].Phytother Res,2014,28(7):961-975.
[7]Jiang M,Yun Q,Niu G,et al.Puerarin prevents inflammation and apoptosis in the neurocytes of a murine Parkinson's disease model[J].Genet Mol Res,2016,15(4):1-9.
[8]Zhou X,Bai C,Sun X,et al.Puerarin attenuates renal fibrosis by reducing oxidative stress induced-epithelial cell apoptosis via MAPK signal pathways in vivo and in vitro[J].Ren Fail,2017,39(1):423-431.
[9]Liu Y,Tang Q,Shao S,et al.Lyophilized Powder of Catalpol and Puerarin Protected Cerebral Vessels from Ischemia by Its Anti-apoptosis on Endothelial Cells[J].Int J Biol Sci,2017,13(3):327-338.
[10]Ma Y,Gai Y,Yan J,et al.Puerarin Attenuates Anoxia/Reoxygenation Injury Through Enhancing Bcl-2 Associated Athanogene 3 Expression,a Modulator of Apoptosis and Autophagy[J].Med Sci Monit,2016,22:977-983.
[11]Agarwala S,Kumar R,Bhatnagar V,et al.High incidence of adriamycin cardiotoxicity in children even at low cumulative doses:role of radionuclide cardiac angiography[J].J Pediatr Surg,2000,35(12):1786-1789.
[12]Cao Y,Shen T,Huang X,et al.Astragalus polysaccharide restores autophagic flux and improves cardiomyocyte function in doxorubicin-induced cardiotoxicity[J].Oncotarget,2017,8(3):4837-4848.
[13]Gan M,Yin X.Puerarin induced in mantle cell lymphoma apoptosis and its possible mechanisms involving multi-signaling pathway[J].Cell Biochem Biophys,2015,71(1):367-373.
[14]Xing G,Dong M,Li X,et al.Neuroprotective effects of puerarin against beta-amyloid-induced neurotoxicity in PC12 cells via a PI3K-dependent signaling pathway[J].Brain Res Bull,2011,85(3-4):212-218.
[15]李聪,周捷,黄海潮,等.葛根素对表柔比星致H9c2心肌细胞凋亡的影响[J].中国药房,2016(34):4807-4810.