定心方经PI3K/Akt通路抑制人脐静脉内皮细胞迁移的研究  

Serum Containing Dingxin Recipe Inhibits Migration of HUVEC through Regulating PI3K/Akt Signal Pathway

作　　者：苏志杰[1] 赵丹丹[1] 周凤华[1] 程赛博[1] 徐煜凌 刘晓瑜[1] 张蕾[1] 张宇[1] 贾钰华[1] SU Zhijie;ZHAO Dandan;ZHOU Fenghua;CHENG Saibo;XU Yuling;LIU Xiaoyu;ZHANG Lei;ZHANG Yu;JIA Yuhua(College of Traditional Chinese Medicine Southern Medical Uuiversity, Guangzhou 510515,Guangdong,China)

机构地区：[1]南方医科大学中医药学院

出　　处：《辽宁中医杂志》2018年第12期2667-2670,2704,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81373574);国家自然科学基金青年基金(81403339);广东省自然科学基金博士启动项目(2014A030310150),广东省自然科学基金项目(2014A030313354)

摘　　要：目的:探讨定心方(DXR)对氧化低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVEC)迁移的分子机制。方法:将HUVEC分为空白组、模型组、DXR低、中、高剂量组和LY294002阻制剂组。除正常对照组外,其他组采用ox-LDL(终浓度为100mg/L)造成HUVEC损伤模型,用不同浓度定心方含药血清(终浓度为0%,5%,10%,20%)和PI3K特异性抑制剂LY294002(浓度为4μmol/L)干预培养24h。采用MTT法检测细胞增殖;细胞划痕实验和迁移实验检测各组细胞迁移情况;Western blotting法检测各组细胞中MMP9、PI3K、p-PI3K、Akt、p-Akt蛋白表达情况。结果:与空白组比较,模型组可显著促进HUVEC增殖和细胞迁移,上调HUVEC内p-PI3K、p-Akt、MMP9蛋白的表达(P<0.05);与模型组比较,DXR高、中剂量组和LY294002阻制剂组可显著抑制HUVEC增殖,降低细胞迁移数目,抑制HUVEC内p-PI3K、p-Akt、MMP9蛋白的表达(P<0.05)。结论:体外实验表明,DXR可抑制ox-LDL诱导的HUVEC迁移,其机制可能与下调PI3K/Akt通路抑制MMP9表达有关。Objective:To investigate the effects of Dingxin Recipe(DXR) in protecting human umbilical vein endothelial cells with ox-LDL-induced injury from migrating and explore the possible mechanism.Methods:Human umbilicalvein endothelial cells(HUVECs) were cultured in vitro and divided into six groups:normal control,oxidative stress injury model,PI3 K/Akt inhibitor group and low,medium and high dose DXR groups.Cell viability and cell migration were analyzed by MTT assay,scratch healing assay,transwell membrane assay and the expressions of MMP9,p-PI3 K and p-Akt proteins were determined by Western blot respectively after exerting into different concentrations f DXR(0,5%,10%,20%) and PI3 K/Akt inhibitor(4 μmol/L).Results:Compared with the control group,the model group could significantly promote the proliferation,scratch-healing and migration of HUVEC,up-regulate the expressions of p-PI3 K,p-Akt and MMP9 ratio in HUVEC.Compared with the model group,10% and 20% of Dingxin Recipe could significantly reduce the proliferation,scratch-healing and migration of HUVEC and down-regulate the expressions of p-PI3 K,p-Akt and MMP9 ratio in HUVEC.Conclusion:Dingxin Recipe could alleviate the cell migration in HUVEC induced by ox-LDL and the mechanism is related to the inhibition of MMP9 via PI3 K/Akt pathway.

关 键 词：定心方 PI3K/AKT信号通路 MMP9 细胞迁移 

分 类 号：R285.5[医药卫生—中药学]