JNK通路对左归丸含药血清调控MC3T3成骨细胞Runx2 mRNA表达的影响  

Effects of JNK Pathway on Runx2 mRNA Expression by Contained Zuogui Pill Serum in MC3T3 Cell

作　　者：刘立萍[1] 任艳玲[1] 李然[1] 赵金茹[2] 蒿长英[1] 宋囡[1] 

机构地区：[1]辽宁中医药大学基础医学院,沈阳110847 [2]辽宁中医药大学实验中心,沈阳110847

出　　处：《中国实验方剂学杂志》2012年第14期143-146,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(30873226);教育部高等学校博士学科点专项科研基金(20102133110001);辽宁省高等学校优秀人才支持计划项目(LR201025)

摘　　要：目的:探讨c-Jun氨端激酶(JNK)信号通路在左归丸含药血清调控成骨前体细胞(MC3T3-E1)增殖和成骨特异转录因子核心结合因子(Runx2)mRNA表达中的作用。方法:以MC3T3-E1为研究对象,制备左归丸含药血清,选用JNK特异抑制剂SP 600125,实验分为空白对照组、SP 600125组、左归丸组、左归丸加SP 600125组、倍美力组、倍美力加SP 600125组。孵育48 h后,采用噻唑蓝(MTT)法检测SP600125对左归丸含药血清干预MC3T3-E1成骨前体细胞增殖作用的影响,采用Western blot法分析JNK蛋白磷酸化水平,采用Real Time RT-PCR法检测成骨细胞特异转录因子Runx2 mRNA表达情况。结果:与空白对照组比较,左归丸含药血清组显著促进细胞增殖,明显上调p-JNK蛋白和Runx2 mRNA表达(P<0.01);SP600125显著抑制左归丸含药血清诱导的增殖和p-JNK蛋白表达(P<0.01),对Runx2 mRNA表达的影响不显著。结论:JNK信号通路的激活可能参与了左归丸含药血清诱导的MC3T3-E1成骨前体细胞增殖,但左归丸含药血清诱导的Runx2mRNA高表达对JNK信号通路依赖不显著。Objective：To examine the role of c-Jun N-terminal kinase（JNK）in proliferation and bone-specific transcription factor runt-related transcription factor 2（Runx2） mRNA expression stimulated by the serum of Zuogui pill in MC3T3-E1 osteoblastic cells.Method： We used model of MC3T3-E1 osteoblastic cells cultured in different serums with or without specific inhibitors of JNK SP 600125 within 48 hours.Grouping：control group,SP 600125 group,Zuogui pill group,Zuogui pill plus SP 600125 group,premarin group,premarin plus SP600125 group.We investigated proliferation using thiazolyl blue（MTT）assay.We analyzed JNK,and p-JNK by immunoblotting.We examined Runx2 mRNA expression in MC3T3-E1 osteoblastic cells by Real Time RT-PCR.Result： Compared with the control group,Zuogui pill group was found to stimulate proliferation significantly,the expression of p-JNK protein and Runx2 mRNA of the Zuogui pill group were increased significantly（P0.01）.SP 600125 decreased the proliferation and expression of p-JNK induced by Zuogui pill（P0.01）,while the expression of Runx2 mRNA was not altered significantly.Conclusion： JNK signaling pathway may be involved in the proliferation induced by Zuogui pill in MC3T3-E1 osteoblastic cells,while Runx2 mRNA expression induced by Zuogui pill was independent of JNK signaling.

关 键 词：左归丸 MC3T3-E1 JNK信号通路 核心结合因子 

分 类 号：R285.5[医药卫生—中药学]