中国中药杂志

2021, v.46(19) 5096-5102

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冠心止痛胶囊阻断MAPK信号通路改善棕榈酸诱导血管内皮细胞损伤的研究
Guanxin Zhitong Capsules attenuate human endothelial cell damage induced by palmitic acid via MAPK signaling pathway

王晓丽;邵静;吴冠信;张薇;周红艳;李开言;孙为;
WANG Xiao-li;SHAO Jing;WU Guan-xin;ZHANG Wei;ZHOU Hong-yan;LI Kai-yan;SUN Wei;Henan Academy of Chinese Medicine;the First Affiliated Hospital of Henan University of Chinese Medicine;

摘要(Abstract):

为了研究冠心止痛胶囊(GXZT)对血管内皮细胞脂毒性损伤的影响,探索GXZT防治动脉粥样硬化可能的作用机制,该实验用棕榈酸(palmitic acid, PA)刺激人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVEC)复制脂毒性损伤模型,将细胞分为正常对照组(NC,15%正常血清)、模型组(PA,0.6 mmol·L~(-1) PA+15%正常血清)、GXZT高剂量组(GXZT-H,0.6 mmol·L~(-1) PA+15%GXZT血清)、中剂量组(GXZT-M,0.6 mmol·L~(-1) PA+10%GXZT血清+5%正常血清)和低剂量组(GXZT-L,0.6 mmol·L~(-1) PA+5%GXZT血清+10%正常血清)。用CCK-8法检测不同组HUVEC的细胞活力,流式细胞术检测细胞凋亡,JC-1荧光探针标记后用激光共聚焦显微镜检测线粒体膜电位(mitochondrial membrane potential, MMP),Western blot检测丝裂原激活蛋白激酶(mitogen-activated protein kinases, MAPK)信号通路关键蛋白p38、ERK1/2、JNK1/2总蛋白和磷酸化蛋白表达,并计算蛋白的磷酸化水平。实验结果显示,与NC组相比,PA组的细胞活力和MMP显著降低(P<0.01,P<0.01),细胞凋亡率显著增加(P<0.01),MAPK信号通路p38、ERK1/2、JNK1/2的磷酸化水平显著增加(P<0.01,P<0.01,P<0.01)。与PA组相比,GXZT-H、GXZT-M、GXZT-L的细胞活力和MMP均显著升高(P<0.01,P<0.01,P<0.01),细胞凋亡率显著降低(P<0.01),p38、ERK1/2、JNK1/2 MAPK信号通路的磷酸化蛋白表达减少,磷酸化水平显著降低(P<0.01,P<0.01,P<0.01)。综上所述,GXZT 3个剂量组均可改善PA诱导的HUVEC脂毒性损伤,其机制可能是通过阻断MAPK信号通路减少内皮细胞凋亡而发挥作用。
The present study observed the effect of Guanxin Zhitong Capsules(GXZT) on the lipotoxicity of vascular endothelial cells and investigated the mechanism of GXZT in atherosclerosis treatment. The lipotoxicity model in human umbilical vein endothelial cells(HUVECs) was induced by palmitic acid(PA) stimulation. These cells were divided into a normal control group(NC, 15% normal serum), a model group(PA, 0.6 mmol·L~(-1) PA+15% normal serum), a high-dose GXZT group(GXZT-H, 0.6 mmol·L~(-1) PA+15% GXZT-medicated serum), a medium-dose GXZT group(GXZT-M, 0.6 mmol·L~(-1) PA+10% GXZT-medicated serum+5% normal serum) and a low-dose GXZT group(GXZT-L, 0.6 mmol·L~(-1) PA+5% GXZT-medicated serum+10% normal serum). HUVECs were detected for cell viability by cell counting kit-8(CCK-8) assay, apoptosis by flow cytometry, mitochondrial membrane potential(MMP) by JC-1 labeled laser scanning confocal microscopy, and total and phosphorylated proteins of p38, ERK1/2, and JNK1/2 in the mitogen-activated protein kinases(MAPK) signaling pathway by Western blot. The phosphorylated level was calcula-ted. Compared with the NC group, the PA group showed decreased cell viability and MMP(P<0.01, P<0.01), elevated apoptosis(P<0.01), and up-regulated phosphorylated levels of p38, ERK1/2, and JNK1/2(P<0.01, P<0.01, P<0.01). Compared with the PA group, the GXZT-H, GXZT-M, and GXZT-L groups showed increased cell viability and MMP(P<0.01, P<0.01, P<0.01), reduced apoptosis(P<0.01), and down-regulated protein expression and phosphorylated levels of p38, ERK1/2 and JNK1/2 in the MAPK signaling pathway(P<0.01, P<0.01, P<0.01). In conclusion, the results suggest that GXZT functions via blocking MAPK signaling pathway to relieve the damage of HUVECs induced by PA.

关键词(KeyWords): 冠心止痛胶囊;人脐静脉内皮细胞;棕榈酸;MAPK信号通路;动脉粥样硬化
Guanxin Zhitong Capsules;human umbilical vein endothelial cells;palmitic acid;MAPK signaling pathway;atherosclerosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81641147);; 河南省中医药科学研究专项课题(2017ZY2004,2018ZY2124)

作者(Author): 王晓丽;邵静;吴冠信;张薇;周红艳;李开言;孙为;
WANG Xiao-li;SHAO Jing;WU Guan-xin;ZHANG Wei;ZHOU Hong-yan;LI Kai-yan;SUN Wei;Henan Academy of Chinese Medicine;the First Affiliated Hospital of Henan University of Chinese Medicine;

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