p38MAPK通路在内脏脂肪素诱导HUVEC细胞分泌TNF-α的作用及定心方含药血清的影响  

Role of p38 MAPK Signal Pathway in Course of Visfatin Inducing HUVEC to Secrete TNF-α and Effect of Dingxin Recipe Containing Serum

作　　者：崔小冰[1] 周凤华[1] 万强[1] 贾钰华[1] 赵晓山[1] 孙学刚[2] 杨静华[3] 刁建新[2] 孙晓敏[1] 

机构地区：[1]南方医科大学中医药学院,广州510515 [2]南方医科大学中医分子生物实验室,广州510515 [3]南方医科大学南方医院,广州510515

出　　处：《中国实验方剂学杂志》2014年第7期135-139,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072776);广州市白云区科技计划(2010-kz-39);广东高校优秀青年创新人才培育项目(LYM09044);广东省自然科学基金博士启动项目(10451051501004674)

摘　　要：目的:观察相对分子质量为38 kD有丝分裂原活化蛋白质激酶(p38 MAPK)信号通路在内脏脂肪素(visfatin)诱导人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)分泌肿瘤坏死因子-α(TNF-α)作用以及定心方(Dingxin Recipe,DXR)的干预影响。方法:制备DXR含药血清;用不同质量浓度(0,50,100,200μg·L-1)及时间点(0,6,12,24,48 h)的visfatin和终浓度5%的DXR含药血清干预HUVEC,采用四氮唑盐(MTT)比色法检测细胞增殖,酶联免疫吸附测定法(ELISA)测定培养上清液中TNF-α含量,Western blotting方法检测细胞p38 MAPK及p-p38 MAPK蛋白表达。结果:与正常对照组相比,100μg·L-1的visfatin作用24 h能显著抑制细胞增殖,升高细胞上清液中TNF-α含量,增强p-p38 MAPK蛋白表达,差异有统计学意义(P<0.05或P<0.01)。DXR能降低HUVEC培养上清液中TNF-α含量,减弱细胞p-p38 MAPK表达,与visfatin组相比差异有统计学意义(P<0.05或P<0.01)。结论:定心方通过调控p38 MAPK通路保护visfatin诱导的HUVEC损伤。Objective： To observe the role of p38 mitogen-activated protein kinases（MAPK） signal pathway in the course of visfatin inducing human umbilical vein endothelial cell（HUVEC） to secrete tumor necrosis factor-α （TNF-α） and the effect of Dingxin recipe （DXR） containing serum. Method： Rats were treated with DXR to prepare DXR containing serum methyl thiazolyl tetrazolium （MTT） method was used to detect the proliferation of HUVEC and the contents of TNF-α a derived from HUVEC culture supernatant were detected by the method of ELISA and the protein expression of p38 MAPK and p-p38 MAPK in HUVEC lysate were detected by the method of Western blotting incorporated into vistation and final concentration 5% DXR containing blood serum, respectively after exerting into different concentration Risfation（0,50,100,200 μg·L^-1） in different time points（0, 6, 12, 24, 48 h）. Result： Compared with the control group, Visfatin（100 μg·L^-1） could significantly decrease the proliferation of HUVEC and increase the TNF-α contents in HUVEC cultural supernatant. Meanwhile, it could increase p-p38 MAPK protein expression at 24 hours. Compared with the visfatin group, DXR（5%） containing serum could decrease the TNF-α contents in HUVEC cultural supernatant, at the same time, it could restrain p-p38 MAPK expression in HUVEC. Conclusion： The p38 MAPK signal pathway plays an important role in HUVEC injury induced by visfatin, the protective mechanism of DXR containing serum is related to the p38 MAPK signal pathway in endothelial cells.

关 键 词：内脏脂肪素 P38 MAPK 定心方含药血清 肿瘤坏死因子-α 

分 类 号：R285.5[医药卫生—中药学]