葛根素通过JNK通路降低内脏脂肪素诱导VEC分泌TNF-α、hs-CRP和IL-6的研究  

Puerarin reduces the secretion of TNF-α,hs-CRP and IL-6 from VEC induced by visfatin through JNK pathway

作　　者：崔小冰[1] 贾钰华[2] 欧阳平[1] 周凤华[2] 万强[3] 邹志明[1] 刘娟[1] 丁园英[1] 邓远燕[1] 郭钰[1] 刘世钊[1] 

机构地区：[1]南方医科大学中西医结合医院,广东广州510310 [2]南方医科大学中医药学院,广东广州510515 [3]江西省中医院,江西南昌330006

出　　处：《广东药学院学报》2015年第6期803-807,共5页Academic Journal of Guangdong College of Pharmacy

基　　金：广东省中医药局课题(20122083);国家自然科学基金项目(81373574);广东省自然科学基金博士启动项目(2014A030310150)

摘　　要：目的观察c-Jun氨基末端激酶(JNK)通路在内脏脂肪素诱导血管内皮细胞(VEC)分泌肿瘤坏死因子-α(TNF-α)、高敏C反应蛋白(hs-CRP)、白介素-6(IL-6)中的作用及葛根素对其分泌的干预作用。方法体外培养VEC,不同质量浓度(0、50、100、200μg/L)内脏脂肪素不同时间点(0、6、12、24、48 h)作用于VEC,并用100μmol/L葛根素及10μmol/L SP600125分别干预VEC,采用MTT法测VEC增殖,ELISA法测VEC上清液中TNF-α、hs-CRP及IL-6含量;Western blotting法测VEC中JNK及其p-JNK表达。结果与对照组相比,100μg/L内脏脂肪素能显著抑制VEC增殖,升高VEC上清液中TNF-α、hsCRP及IL-6含量且能上调p-JNK蛋白表达;与内脏脂肪素组比较,葛根素及SP600125均能显著减轻内脏脂肪素对VEC增殖的抑制,减少内脏脂肪素诱导VEC上清液中TNF-α、hs-CRP及IL-6的含量并能下调VEC中p-JNK蛋白的表达。结论葛根素通过调控JNK通路保护由内脏脂肪素诱导的VEC损伤。Objective To study the role of JNK pathway in VEC secretion of tumor necrosis factor-α （TNF- α） ,hs-CRP and IL-6 induced by visfatin induction,and the intervention effect of Puerarin. Methods VEC were cultured and added different concentrations of visfatin （0,50,100 and 200 μg/L） at different times （0,6,12,24 and 48 h）. Meantime, puerarin （ 100 μmol/L） and SP600125 （ 10 μmol/L）, the specificinhibitor of JNK pathway, were added into VEC culture. Methyl thiazolyl tetrazolium （MTT） method was used to detect the proliferation of VEC. The contents of TNF-α, hs-CRP and IL-6 derived from VEC supernatant were measured by ELISA. The protein expressions of p-JNK and JNK in VEC were detected by western blotting. Results Compared with the control group, 100 μg/L visfatin significantly decreased the proliferation of VEC and increased the contents of TNF-α, hs-CRP and IL-6 in VEC supernatant. Meanwhile, visfatin increased p-JNK expression at 24 hours. Compared with the visfatin group, puerarin and SP600125 enhanced the proliferation of VEC, decreased the levels of TNF-α, hs-CRP and IL-6 in VEC supernatant, and restrained p-JNK expression in VEC. Conclusion Puerarin reduces VEC injury induced by visfatin through JNK pathway.

关 键 词：内脏脂肪素 葛根素 C-JUN氨基末端激酶 肿瘤坏死因子-α 高敏C反应蛋白 白细胞介素-6 血管内皮细胞 

分 类 号：R285.5[医药卫生—中药学]