摘要
目的探讨芍药苷(PF)对高浓度葡萄糖(HG)诱导H9c2细胞损伤的作用及机制。方法H9c2细胞分别用PF 250,300,350μmol·L^-1、葡萄糖35 mmol·L^-1(HG组)、SB2035803μmol·L^-1、SP60012510μmol·L^-1、U012615μmol·L^-1单独或联合PF 350μmol·L^-1处理3 h,加葡萄糖35 mmol·L^-1继续孵育21 h。CCK-8法检测细胞存活率,流式细胞术检测细胞凋亡,Western印迹法检测活化胱天蛋白酶3、Bax和Bcl-2蛋白表达水平及P38促分裂原活化的蛋白激酶(P38 MAPK)、细胞外信号调节蛋白激酶1/2(ERK1/2)和c-Jun N端激酶激酶(JNK)蛋白磷酸化水平。结果与HG组相比较,HG+PF 300和350μmol·L^-1组、HG+SB2035803μmol·L^-1组、HG+SP60012510μmol·L^-1组、HG+U012615μmol·L^-1组细胞存活率显著升高(P<0.05,P<0.01),细胞凋亡率显著降低(P<0.05,P<0.01),胱天蛋白酶3和Bax蛋白显著下调(P<0.05,P<0.01),Bcl-2蛋白显著上调(P<0.05)。与HG组相比较,HG+PF 300和350μmol·L^-1组P38 MAPK,ERK1/2和JNK磷酸化水平显著下调(P<0.05,P<0.01),HG+SB2035803μmol·L^-1组P38 MAPK磷酸化水平显著下调(P<0.01),HG+SP60012510μmol·L^-1组JNK磷酸化水平显著下调(P<0.01),HG+U012615μmol·L^-1组ERK1/2磷酸化水平显著下调(P<0.01)。结论PF通过抑制MAPK通路活化来抑制HG诱导的H9c2心肌细胞毒性和凋亡,从而减轻HG诱导的H9c2心肌细胞损伤。
OBJECTIVE To investigate the effect and mechanism of paeoniflorin(PF) on H9c2 cell injury induced by high glucose(HG). METHODS H9 c2 cells were treated with PF 250, 300 and 350 μmol·L^-1, glucose 35 mmol·L^-1(HG group), SB203580 3 μmol·L^-1, SP600125 10 μmol·L^-1 and U0126 15 μmol·L^-1 alone or in combination for 3 h, followed by glucose 35 mmol·L^-1 for 21 h. Cell viability was detected by CCK-8 assay, apoptosis was detected by flow cytometry, and the expression levels of cleaved-caspase 3, Bax, and Bcl-2 proteins, and the phosphorylation levels of P38 mitogenactivated protein kinase(P38 MAPK), extracellular signal-regulated protein kinase 1/2(ERK1/2) and c-Jun N-terminal kinase(JNK) proteins were detected by Western blotting. RESULTS Compared with HG group, the survival rate of HG+PF 300 and 350 μmol·L^-1 groups, HG+SB203580 3 μmol·L^-1 group,HG+SP600125 10 μmol·L^-1 group and HG+U0126 15 μmol·L^-1 group were significantly increased(P<0.05, P<0.01), the apoptosis rate was significantly decreased(P<0.05, P<0.01), caspase 3 and Bax were significantly down-regulated(P<0.05, P<0.01), and the expression of Bcl-2 protein was significantly up-regulated(P<0.05). Compared with HG group, the phosphorylation levels of P38 MAPK, ERK1/2 and JNK were significantly down-regulated in HG+PF 300 and 350 μmol·L^-1 groups(P<0.05, P<0.01),the phosphorylation level of P38 MAPK was significantly down-regulated in HG + SB203580 3 μmol·L^-1 group(P<0.01), that of JNK was significantly down-regulated in HG+SP600125 10 μmol·L^-1 group(P<0.01), and that of ERK1/2 was significantly down-regulated in HG+U0126 15 μmol·L^-1 group(P<0.01).CONCLUSION PF can inhibit HG-induced H9 c2 cells toxicity and apoptosis by inhibiting MAPK pathway activation, thereby inhibiting HG-induced damage to H9c2 cells.
作者
王艾青
陈玉芳
申洪娇
杨卓
田振涛
WANG Ai-qing;CHEN Yu-fang;SHEN Hong-jiao;YANG Zhuo;TIAN Zhen-tao(Office of Academic Affairs,Henan Vocational College of Nursing,Anyang 455000,China;Department of General Medicine,Anyang District Hospital of Puyang City,Anyang 455000,China;Department of Traditional Chinese Medicine,Anyang District Hospital of Puyang City,Anyang 455000,China)
出处
《中国药理学与毒理学杂志》
CAS
北大核心
2020年第10期721-728,共8页
Chinese Journal of Pharmacology and Toxicology
基金
河南省医学研究项目(Wjlx2016043)。
