摘要
目的:探讨JNK-c-Jun/AP-1信号转导通路在血管紧张素Ⅱ(AngⅡ)诱导的肾小球系膜细胞(MC)增殖及细胞周期调控中的作用。方法:应用3H-胸腺嘧啶(3H-TdR)掺入法及流式细胞术测定MC增殖和细胞周期的变化。应用凝胶电泳迁移率(EMSA)和非放射性激酶活性检测法检测系膜细胞内活化蛋白-1(AP-1)及c-Jun氨基末端激酶(JNK)活性。结果:AngⅡ可呈时间依赖性地诱导MC内JNK活化,AngⅡ刺激30min后,JNK活性达到高峰,1h几乎恢复至正常水平;AngⅡ刺激后MC内AP-1活性显著增强,3H-TdR掺入量明显增加,S期和G2/M期细胞数显著增多;JNK特异性抑制剂SP600125显著抑制AngⅡ诱导AP-1活化及MC增殖。结论:AngⅡ→JNK/SAPK→c-Jun/AP-1信号通路在MC增殖中发挥一定的作用。JNK特异性抑制剂SP600125能部分抑制AngⅡ诱导的AP-1活化及细胞增殖,从而可能具有一定的治疗作用。
Objective:To investigated the role of JNK-c-Jun/AP-1 signal pat hway in the regulation of angiotensin (Ang) Ⅱon human mesangial cells (MC) prol iferation at the level of the cell-cycle. Methods:The incorporation of 3H-thy midine (3H-TdR) was used as the measure of MC proliferation. MC cell-cycle was analyzed by flow cytometry. Electrophoretic mobility shift assay (EMSA) and non -radioactive kinase assay were used to detect the activity of AP-1 and JNK. Re sults:Ang Ⅱenhanced the activities of JNK in a time-dependent manner, with a peak at 30 min and the activities of JNK returned to the basal levels after 1 h. EMSA showed that Ang Ⅱactivated AP-1 in MC. Ang Ⅱpromtoed a 2.24-fold stimu lation of 3H-TdR incorporation. Significant down-regulation of JNK and AP-1 a ctivation, and MC proliferation were observed in Ang Ⅱ-treated MC pretreated w ith JNK specific inhibitor SP600125. Conclusion:JNK-c-Jun/AP-1 signal pathwa y may play an important role in the proliferation of MC induced by Ang Ⅱ.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2004年第1期4-8,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助项目(30100081)
江苏省教育厅自然科学基金资助项目(02KJD320011
01KJD320009)
