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

杨翼鹰; 陈伟燕; 孙秀亭; 王翔; 李政勋; 梁美玲; 石卉; 杨至圣; 曾武涛

doi:10.13418/j.issn.1001-165x.2016.05.009

中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (5) : 517-522. DOI: 10.13418/j.issn.1001-165x.2016.05.009

实验研究

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

杨翼鹰¹，陈伟燕²，　孙秀亭¹，　王翔³，　李政勋¹，　梁美玲⁴，　石卉¹，　杨至圣¹，　曾武涛¹

作者信息 +

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

YANG Yi-ying¹, CHEN Wei-yan², SUN Xiu-ting¹, WANG Xiang³, LI Zheng-xun¹, LIANG Mei-ling⁴, SHI Hui¹, YANG Zhi-sheng¹, ZENG Wu-tao¹

Author information +

文章历史 +

摘要

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p38 MAPK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、p38 MAPK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。

Abstract

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．

导出引用

杨翼鹰,陈伟燕,孙秀亭,王翔,李政勋,梁美玲,石卉,杨至圣,曾武涛. 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[J]. 中国临床解剖学杂志. 2016, 34(5): 517-522 https://doi.org/10.13418/j.issn.1001-165x.2016.05.009

YANG Yi-ying, CHEN Wei-yan, SUN Xiu-ting, WANG Xiang, LI Zheng-xun, LIANG Mei-ling, SHI Hui, YANG Zhi-sheng, ZENG Wu-tao. Shexiang baoxin pill protects cardiomyocytes against high glucose-induced injury by inhibiting the p38 MAPK and NF-κB pathway[J]. Chinese Journal of Clinical Anatomy. 2016, 34(5): 517-522 https://doi.org/10.13418/j.issn.1001-165x.2016.05.009

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