【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。更多还原

【Abstract】 Objective To determine whether JNK signaling pathway is involved in high glucose(HG)-induced proliferation 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 indicated 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 stimulates the proliferation of CFs partially through JNK pathway activation.更多还原