摘要
目的观察活化蛋白1(AP-1)在高糖诱导MC3T3-E1成骨细胞凋亡中的作用。方法将体外培养的MC3T3-E1细胞分为正常对照组和实验组,实验组包括22.0 mmol/L D-葡萄糖组(高糖组)、P38MAPK-shRNA慢病毒转染组、P38MAPK信号转导阻断剂组、无关shRNA转染组、AP-1抑制剂SP600125组。采用TUNEL和流式细胞术检测细胞凋亡,用EMSA检测MC3T3-E1细胞AP-1的活性。结果与正常对照组相比,高糖组MC3T3-E1成骨细胞P38MAPK表达、AP-1活性、细胞凋亡显著增加。P38MAPK-shRNA慢病毒转染组和P38MAPK信号转导阻断剂组MC3T3-E1细胞AP-1活性较高糖组分别下降57.9%(P<0.01)和45.6%(P<0.05)。AP-1抑制剂组MC3T3-E1细胞凋亡率较高糖组下降39.7%(P<0.01)。结论高糖通过激活P38MAPK增加AP-1活性,诱导MC3T3-E1成骨细胞凋亡,抑制AP-1活性后高糖诱导的MC3T3-E1细胞凋亡显著下降。
Objective To investigate the effect of AP-1 on the apoptosis of MC3T3-E1 osteoblast induced by high-glucose.Methods MC3T3-E1 cells cultured in vitro were divided into normal control group and treated groups that including 22.0 mmol/L D-glucose group, P38MAPK-shRNA transfection group, P38MAPK signal transduction in-hibitor group, transfection with negative control siRNA group and AP-1 inhibitor SP600125 group. Apoptosis of MC3T3-E1 cell was measured with TUNEL and flow cytometry;activities of activator protein-1 （ AP-1） were measured with EMSA.Results Compared with normal glucose group, P38MAPK expression, AP-1 activity and MC3T3-E1 cell apoptosis were increased obviously in high-glucose group.After P38MAPK-shRNA transfection and use of P38MAPK inhibitor, AP-1 activity in MC3T3-E1 cell decreased by 57.9% （P〈0.01） and 45.6% （P〈0.05） respectively. The apoptosis of MC3T3-E1 cell in AP-1 inhibitor group decreased by 39.7% comparing to high-glucose group （ P〈0.01）.Conclusion High-glucose induces the apoptosis of MC3T3-E1 cell through activation of P38MAPK and subse-quently activation of AP-1, and decreased activity of AP-1 can ameliorate the apoptosis of MC3T3-E1 cell induced by high-glucose.
出处
《中华骨质疏松和骨矿盐疾病杂志》
2014年第3期258-262,共5页
Chinese Journal Of Osteoporosis And Bone Mineral Research
基金
国家临床重点专科建设项目
重庆市卫生局科研基金(09-02-053)
