基于ERK/p38 MAPK信号通路探讨芍药苷对顺铂诱导大鼠心功能障碍及心肌细胞损伤的影响

Study of the Effects of Paeoniflorin on Cardiac Dysfunction and Myocardial Cell Injury Induced by Cisplatin Based on ERK/p38 MAPK Signaling Pathway

关键词:  心功能障碍  PF  炎症  凋亡  ERK/p38 MAPK

Key words:  cardiac dysfunction  PF  Inflammation  Apoptosis  ERK/p38 MAPK

摘要:

目的 探究芍药苷(paeoniflorin，PF)对顺铂(cisplatin,CDDP)诱导大鼠心功能障碍及心肌细胞损伤的保护作用。方法 SD雄性大鼠随机分为对照组(CN组)、CDDP组、CDDP+PF低剂量干预组(CDDP+PF1组)及CDDP+PF高剂量干预组(CDDP+PF2组)，Powerlab多功能记录仪进行左心室插管检测心功能相关指标：左室内压峰值(LVSP)、左室舒张末压(LVEDP)、左室内压变化速率(±dp/dt)的数值变化；取各组大鼠血清，测炎性因子TNF-α、IL-1β和IL-6水平；取心肌组织染色，观察组织结构变化。H9c2心肌细胞分为对照组(CN组)、CDDP组、PF组及CDDP+PF组，CCK-8测各组细胞活性；流式细胞术检测各组心肌细胞凋亡情况；蛋白免疫印迹法(Western Blot)检测心肌细胞中MAPK信号通路相关蛋白p38、ERK、JNK及其磷酸化蛋白表达和凋亡相关蛋白Bax、Bcl-2、Casp3、Cl-casp3的表达情况。结果 与CN组相比，CDDP组LVSP(P < 0.01)、±dp/dt (P < 0.01)下降，LVEDP升高 (P < 0.01)；PF低剂量和高剂量预处理均可升高LVSP(P < 0.01)及±dp/dt数值 (P < 0.01)、降低LVEDP (P < 0.01)；且PF低剂量和高剂量预处理均可降低大鼠血清中炎性因子TNF-α(P < 0.01)、IL-1β(P < 0.01)和IL-6 (P < 0.01)含量。细胞水平结果显示：与CN相比，CDDP组细胞活性下降，凋亡相关蛋白Bax (P < 0.01)、Cl-casp3(P < 0.01)表达增加，抗凋亡蛋白Bcl-2 (P < 0.01)表达降低，MAPK通路p38及ERK磷酸化表达升高；与CDDP组相比，PF可恢复细胞活性，下调凋亡相关蛋白Bax (P < 0.01)、Cl-casp3 (P < 0.01)表达，增加抗凋亡蛋白Bcl-2 (P < 0.01)表达，抑制MAPK通路p38 (P < 0.01)及ERK (P < 0.01)磷酸化表达。结论 PF可恢复CDDP诱导大鼠心功能障碍及心肌细胞损伤，其作用可能通过调控ERK/p38 MAPK信号抑制炎症反应和细胞凋亡。

Abstract:

ABSTRACT: OBJECTIVE To investigate the protective effect of paeoniflorin on cardiac dysfunction and myocardial cell injury induced by cisplatin in rats. METHODS SD male rats were randomly divided into control group, cisplatin group, cisplatin + paeoniflorin low-dose intervention group and cisplatin + paeoniflorin high-dose intervention group. Powerlab multifunctional recorder was used to detect the related indexes of cardiac function: The changes of left ventricular peak pressure (LVSP), left ventricular end-diastolic pressure (LVDEP) and left ventricular pressure change rate (±dp/dt); Serum levels of inflammatory factors TNF-α, IL-1β and IL-6 were measured in each group. Myocardial tissue was stained to observe the changes of tissue structure. H9c2 cardiomyocytes were divided into control group, cisplatin group, paeoniflorin group and cisplatin + paeoniflorin group. The activity of H9C2 cardiomyocytes was measured by CCK-8. The apoptosis of cardiomyocytes in each group was detected by flow cytometry. The expressions of MAPK signaling pathway related proteins p38, ERK, JNK and their phosphorylated proteins and apoptosis-related proteins Bax, Bcl-2, casp3、cl-casp3 were detected in cardiomyocytes by Western Blot. RESULTS Compared with the control group, LVSP (P < 0.01), ±dp/dt (P < 0.01) decreased, LVDEP increased (P < 0.01); Both low-dose and high-dose PF pretreatment increased LVSP (P < 0.01) and ±dp/dt value (P < 0.01), decreased LVDEP (P < 0.01); Both low-dose and high-dose PF pretreatment could decrease the serum inflammatory factor TNF-α(P < 0.01), IL-1β(P < 0.01) and IL-6 (P < 0.01). Cell level results showed that compared with control group, CDDP group decreased cell activity and apoptosis-related protein BAX (P < 0.05), cl-casp3 (P < 0.01) increased expression of anti-apoptotic protein bcl-2 (P < 0.05). And the expression of p38 and ERK phosphorylation also increased; Compared with CDDP group, PF can restore cell activity and down-regulate apoptosis-related protein BAX (P < 0.01), cl-casp3 (P < 0.01) and increased anti-apoptotic protein bcl-2 (P < 0.01) expression, inhibit MAPK pathway p38 (P < 0.01) and ERK (P < 0.01) phosphorylation expression. CONCLUSION PF can restore cardiac dysfunction and myocardial cell injury induced by cisplatin in rats, which may be related to inhibiting inflammation and apoptosis by regulating ERK/p38 MAPK signal expression.