左、右归丸含药血清通过p38MAPK信号通路干预BMSCs成骨诱导的研究  

Effects of Serum Containing Zuogui Pill,Yougui Pill and Their Disassembled Prescriptions on Osteogenic Differentiation of BMSCs through p38 Pathway

作　　者：曲宁宁[1] 何丽娟[1] 何文智[1] 任艳玲[1] 

机构地区：[1]辽宁中医药大学基础医学院,辽宁沈阳110847

出　　处：《中华中医药学刊》2016年第6期1326-1329,I0005,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81373527)

摘　　要：目的:基于p38信号通路探讨左、右归丸含药血清对BMSCs成骨诱导的机制。方法:运用全骨髓贴壁法分离和培养大鼠BMSCs;分别以左归丸、右归丸、两方共同药、滋肾阴药、补肾阳药、阳性对照药补佳乐制备的大鼠含药血清加诱导剂(地塞米松、维生素C、β-甘油磷酸钠)、诱导剂和空白含药血清组共8组对BMSCs进行干预,采用改良钙钴染色法检测碱性磷酸酶(ALP)表达,采用茜素红染色法检测钙化结节,采用Western blotting法检测核结合因子α1(Cbfα1)和Ⅰ型胶原(ColⅠ)、p38、p-p38蛋白表达,采用real time PCR法检测Cbfα1、ColⅠmRNA表达。结果:左、右归丸及滋阴药组可以上调ALP表达,促进BMSCs矿化结节形成,增强Cbfα1、ColⅠmRNA和蛋白的表达并且可以促进p38蛋白的磷酸化;给予p38特异性阻滞剂SB203580后,各组BMSCs ALP表达下调,矿化结节形成减少,p38蛋白磷酸化水平降低,并且Cbfα1、ColⅠmRNA和蛋白的表达下降。结论:左、右归丸及其拆方含药血清可能部分通过p38 MAPK信号通路对BMSCs成骨分化产生调控作用的。Objective： To explore the mechanism of serum containing Zuogui Pill and Yougui Pill on BMSCs through p 3 8 signaling pathway. Methods： The whole bone marrow adherence method was used to isolate and culture BMSCs.Zuogui Pill,Yougui Pill,combined formula,nourishing Yin drugs,supplementing kidney Yang drugs,positive control progynova model rats＇ serum plus inducers（ dexamethasone,vitamin C,β-glycerophosphate）,inducers and blank serum,all together 8 groups of BMSCs intervention. The modified calcium cobalt staining was used to detected alkaline phosphatase（ ALP） expression and alizarin red staining for calcified nodules and Western blotting for nuclear binding factor α1（ Cbfα1）,collagen Ⅰ（ ColⅠ）,p38 and p-p38 protein expression and real time PCR method for detecting Cbfα1 and ColⅠmRNA expression. Results： Zuogui Pill,Yougui Pill and nourishing Yin drug groups can increase ALP expression,promote BMSCs mineralized nodule formation and enhance Cbfα1,ColⅠ mRNA and protein expression and phosphorylation of p38 protein and after giving specific p38 sexual blocker SB203580,ALP expression in each group BMSCs was down,the formation of mineralized nodules reduced and p38 protein phosphorylation lower and Cbfα1,ColⅠ mRNA and protein expression decreased. Conclusion： Serum Containing Zuogui Pill and Yougui Pill and its decomposed formulae may be partially through p38 MAPK signaling pathway on osteogenic differentiation of BMSCs produce regulatory role.

关 键 词：左归丸 右归丸 骨髓间充质干细胞 P38 成骨诱导 

分 类 号：R285.5[医药卫生—中药学]