摘要
目的明确C-JUN氨基末端激酶(JNK)信号通路在高糖诱导大鼠心肌成纤维细胞增殖中的作用。方法提取大鼠原代心肌成纤维细胞,观察不同糖浓度(12、18和25 mmol/L葡萄糖)和不同刺激时间(0、12、24和48 h)对JNK通路的影响。实验分为对照组(5.5 mmol/L葡萄糖)、高糖组(25 mmol/L葡萄糖)、JNK抑制剂SP600125组(25 mmol/L葡萄糖+SP600125 10、20μmol/L)和高渗组(5.5 mmol/L葡萄糖+19.5 mmol/L甘露醇)培养48 h,应用MTT测定细胞增殖、Weastern blot测定JNK磷酸化水平和增殖细胞核抗原(PCNA)的蛋白表达、RT-PCR检测C-JUN基因表达。结果高糖呈时间和剂量依赖性增加JNK磷酸化水平。与对照组比较,高糖显著地促进了心肌成纤维细胞的增殖(0.44±0.02 vs 0.31±0.02,P<0.01),并上调了C-JUN的基因表达和PCNA蛋白水平。SP600125能够浓度依赖性地抑制高糖诱导的细胞增殖和JNK通路的激活。结论 JNK信号通路部分地介导了高糖诱导的大鼠心肌成纤维细胞增殖。
Objective To determine whether JNK signaling pathway is involved in high glucose （HG） -induced pro- liferation of cardiac fibroblasts （CFs） in rats. Methods Rat CFs were cultured in DMEM （ NG： 5.5 mmol/L D-glucose; HG： 12, 18, 25 mmol/L D-glucose; OSM： 5.5 mmol/L D-glucose ＋ 19. 5 mmol/L mannitol） for in- dicated time periods（0, 12, 24, 48 h）. Meanwhile, rat CFs were cultured in DMEM （25 mmol/L D-glucose） with SP600125（ 10, 20 μmol/L）, a JNK inhibitor, for 48 h. Proliferation was measured by MTT. The expression of proliferating cell nuclear antigen （PCNA）, total JNK and phosphorylation of JNK were detected by Western blot. C-JUN mRNA expression was assessed by RT-PCR. Results Treatment of CFs with HG significantly increased phosphorylation of JNK in time-and glucose concentration-dependent manner. Compared with NG group, exposure of CFs to HG （ 25 mmol/L） significantly promoted the proliferation of CFs （ 0. 44 ±0. 02 vs 0. 31 ± 0. 02, P 〈 0. 01）, and upregulated C-JUN mRNA expression and PCNA levels. SP600125 significantly suppressed HG-induced the proliferation of CFs and the activity of JNK in a dose-dependent manner. Conclusion HG stimulatesthe proliferation of CFs partially through JNK pathway activation.
出处
《基础医学与临床》
CSCD
北大核心
2013年第2期199-204,共6页
Basic and Clinical Medicine
基金
河北省卫生厅重大课题资助项目(20090007)
