黄连素通过ERK1/2通路降低内脏脂肪素诱导HUVEC分泌TNF-α和IL-6的研究  

Berberine Reduce TNF-α and IL-6 Secreted in HUVEC Induced by Visfatin through ERK1 /2 Signal Pathway

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

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14期125-129,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072776);广州市海珠区科技计划项目(2013-cg-35)

摘　　要：目的:观察细胞外调节蛋白激酶(ERK1/2)通路在内脏脂肪素(visfatin)诱导人脐静脉内皮细胞(HUVEC)分泌肿瘤坏死因子-α(TNF-α)及白介素-6(IL-6)中的作用及黄连素(berberine,Ber)的干预作用。方法:体外培养HUVEC,用visfatin不同质量浓度(0,50,100,200μg·L-1)、不同时间(0,6,12,24,48 h)作用HUVEC,MTT法测HUVEC增殖,ELISA法测HUVEC上清液中TNF-α及IL-6含量;Western blotting法测HUVEC中p-ERK1/2蛋白表达;并用50μmol·L-1Ber及ERK1/2通路特异性抑制剂PD98059 20μmol·L-1干预HUVEC,检测Ber的干预作用。结果:与对照组相比,100μg·L-1visfatin能显著抑制HUVEC增殖,升高HUVEC上清液中TNF-α及IL-6含量并能上调p-ERK1/2蛋白表达;与visfatin组比,Ber及PD98059均能显著减轻visfatin对HUVEC增殖的抑制,减少visfatin诱导HUVEC上清液中TNF-α及IL-6的含量并能下调HUVEC中pERK1/2蛋白的表达。结论:Ber可通过抑制炎症因子TNF-α及IL-6的分泌,减轻visfatin诱导的HUVEC损伤,其机制可能与抑制ERK1/2通路的激活有关。Objective： To study the role of extra cellular regulated protein kinasses （ERK1/2） pathway in tumor necrosis factor-alpha （TNF-α） and interleukin-6 （IL-6） secreted in human umbilical vein endothelial cell （HUVEC） induced by visfatin and the interventive effect of berberine. Method： HUVEC were cultured for the experiment, added concentration gradient group of visfatin （0, 50, 100, 200μg·L^-1） and time gradient group of visfatin （0, 6, 12, 24, 48 h） in HUVEC, MTT assay was used to detect the proliferation of HUVEC,the contents of TNF-α and IL-6 in HUVEC supernatant were determined by ELISA assay, the expression of p- ERK1/2 in HUVEC lysate were determined by Western blotting. Berberine （50 μmol ·L^-1） and PD98059 （20 μmol · L ^-1 ） , the specific inhibitor of ERK1/2 pathway, were added into HUVEC to detected the interventive effect of berberine. Result： Compared with the control group, visfatin （100 μg·L^-1） can significantly decrease the proliferation of HUVEC and increase the TNF-α and IL-6 contents in HUVEC supernatant. Meanwhile, it can increase p-ERK1/2 expression at 24 hours. Compared with the visfatin group, berberine and PD98059 could increase the proliferation of HUVEC and decrease the TNF-α and IL-6 contents in HUVEC supernatant, at the same time, it could restrain p-ERK1/2 expression in HUVEC. Conclusion： Berberine can decrease the contents of TNF-α and IL-6 in HUVEC cultural supernatant and the injury induced by visfatin, the protective mechanism is related to the inhibition of ERK1/2 signal pathway activation.

关 键 词：内脏脂肪素 细胞外调节蛋白激酶 黄连素 肿瘤坏死因子-α 白细胞介素-6 

分 类 号：R285.5[医药卫生—中药学]