麝香保心丸通过抑制p38 MAPK和NF-κB信号传导通路表达对抗高糖诱导的心肌细胞损伤

Shexiang baoxin pill protects cardiomyocytes against high glucose-induced injury by inhibiting the p38 MAPK and NF-κB pathway

目的探讨麝香保心丸(SBP)是否通过抑制p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)和核因子kappa B(nuclear factor kappa B,NF-κB)通路保护H9c2心肌细胞对抗高浓度葡萄糖(高糖,HG)引起的损伤。方法应用35 mmol/L的HG处理H9c2心肌细胞24 h,建立HG诱导的心肌细胞损伤模型;细胞计数试剂盒8测定细胞存活率;Hoechst 33258核染色荧光显微镜照相法测定细胞凋亡;双氯荧光素(2’,7’-dichlorfluorescein-diacetate,DCFH-DA)染色/荧光显微镜照相法测定胞内活性氧(ROS)水平;罗丹明123(Rh123)染色法测定线粒体膜电位(MMP);Western blot法测定p38MAPK和NF-κB p65蛋白表达水平。结果 HG处理H9c2心肌细胞24 h能引起细胞明显的损伤,使细胞存活率降低,凋亡细胞数量和细胞内ROS生成增多,MMP丢失;HG能增加p38 MAPK和NF-κB p65磷酸化水平;SBP预处理能明显抑制HG上调p38 MAPK和NF-κB p65磷酸化水平这一作用;SBP、p38MAPK通路抑制剂SB203580和NF-κB通路抑制剂PDTC均能阻断HG对心肌细胞的上述损伤作用,包括细胞毒性、凋亡、ROS生成增多及MMP丢失等。结论麝香保心丸(SBP)可通过抑制p38 MAPK和NF-κB信号传导通路保护H9c2心肌细胞对抗高糖(HG)引起的损伤。

Objective To explore whether Shexiang Baoxin Pill（SXBXP/SBP）protects H9c2 cardiac cells against high glucose（HG）-induced injury by inhibition of p38 MAPK and NF-κB pathway. Methods H9c2 cardiac cells were treated with 35 mmol/L glucose（HG）for 24h to establish a model of HG-induced injury;Cell counter kit-8（CCK-8）was used to measure cell viability;Apoptotic cells were tested by Hoechst 33258 nuclear staining followed by fluorescence imaging;Intracellular levels of reactive oxygen species （ROS）were detected by 2’,7’-dichlorfluorescein-diacetate（DCFH-DA）staining,followed by fluorescence imaging; Mitochondrial membrane potential （MMP）was measured by a fluorescent dye,rhodamine 123 （Rh123）,followed by fluorescence imaging;The expression levels of p38 MAPK and NF-κB p65 protein were detected by Western blot assay. Result Treatment of H9c2 cardiac cells with HG for 24h signally induced injuries,proved by a decrease in cell viability,an increase in amount of apoptotic cells and intracellular ROS production,as well as a loss of MMP;HG strengthened the expression of phosphorylated （p）-p38 MAPK and p-NF-κB p65;Pretreatment with SBP notably inhibited the up-regulation of expression of p-p38 MAPK and p-NF-κB p65 induced by HG; Pretreatment with SBP or SB203580,an inhibitor of p38 pathway or PDTC, an inhibitor of NF-κB pathway,attenuated the above HG-induced injuries,including cytotoxicity,apoptosis,increase in ROS production and a loss of MMP,in H9c2 cardiac cells. Conclusion SXBXP（SBP）may protect H9c2 cardiac cells against the HG-induced injuries by inhibiting the p38 MAPK and NF-κB pathway.