左归丸含药血清通过p38 MAPK信号通路诱导MC3T3-E1成骨细胞的分化  

Effects of p38 signal pathway on osteoblastic cell differentiation by Zuogui pill medicated serum in MC3T3-E1 cell

作　　者：刘立萍[1] 任艳玲[1] 李然[1] 王智民[2] 赵金茹[3] 蒿长英[1] 宋囡[1] 

机构地区：[1]辽宁中医药大学基础医学院,辽宁沈阳110847 [2]辽宁中医药大学第一临床学院 [3]辽宁中医药大学实验中心

出　　处：《中国老年学杂志》2013年第7期1579-1581,共3页Chinese Journal of Gerontology

基　　金：国家自然基金项目(No.30873226);教育部高等学校博士学科点专项科研基金(No.20102133110001);辽宁省高等学校优秀人才支持计划资助(No.LR201025);辽宁省教育厅创新团队项目(No.LT2010068)

摘　　要：目的探讨p38 MAPK信号通路对左归丸含药血清干预MC3T3-E1成骨细胞分化的影响。方法制备大鼠含药血清,选用p38特异阻滞剂SB203580,实验分为空白对照组、SB203580组、左归丸组、左归丸加SB203580组、倍美力组、倍美力加SB203580组。孵育48 h后,采用PNPP法检测碱性磷酸酶(ALP)活性,采用Western印迹法分析ALP蛋白表达水平;孵育7 d,采用改良钙钴染色法检测ALP活性;孵育14 d,采用茜素红染色法测定矿化沉积情况。结果与空白对照组比较,左归丸组显著促进MC3T3-E1成骨细胞分化,明显上调ALP蛋白表达水平(P<0.01);与左归丸组比较,左归丸加SB203580组显著抑制左归丸含药血清对MC3T3-E1成骨细胞的分化的促进作用,明显下调ALP蛋白表达水平(P<0.01)。结论左归丸含药血清可能部分通过p38 MAPK信号通路干预MC3T3-E1成骨细胞分化。Objective To investigate the role of p38 MAPK in osteoblastic call differentiation stimulated by the serum of Zuogui pill in MC3T3-E1 osteoblastic cells. Methods Cells were cultured in different rat serums with or without specific inhibitors of p38 SB203580. Cells were divided into control, SB203580, Zuogui pill, Zuogui pill plus SB203580, Premarin, Premarin plus SB203580 groups. Alkaline phosphatase （ALP） activity was investigated by using PNPP method and the expression of ALP protein was analyzed by immunoblotting with- in 48 h. ALP staining was used to detect ALP activity within 7 d and alizarin red staining to detect mineralized nodules within 14 d. Results Compared with the control group, Zuogui pill could Stimulate ALP activity, expression of ALP protein, and mineralized nodules significant- ly （P 〈 0. 01 ）. Compared with the Zuogui pill group, the ALP activity and mineralized nodules treated with Zuogui pill plus SB203580 were decreased significantly （P 〈0. 01 ）. Conclusions Zuogui pill regulates MC3T3-E1 osteoblastic cell differentiation through the activation of p38 MAPK signal transduction pathways.

关 键 词：左归丸 成骨细胞 P38MAPK信号通路 碱性磷酸酶 矿化沉积 

分 类 号：R285[医药卫生—中药学]