左、右归丸对大鼠骨髓间充质干细胞骨向分化中TGF-β_1 mRNA的影响  

Effect of Zuogui Wan and Yougui Wan on TGF-β_1 on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

作　　者：宋囡[1] 何文智[2] 王智民[3] 任艳玲[1] 

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

出　　处：《中国实验方剂学杂志》2013年第18期210-214,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81373527);辽宁省教育厅创新团队项目(LT2010068);辽宁省高等学校优秀人才支持计划(LR201025);辽宁省科技厅自然科学基金项目计划(201102148)

摘　　要：目的:研究左、右归丸含药血清对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)成骨分化的作用。方法:将28只大鼠随机分为4组,分别以左归丸(ZGW)、右归丸(YGW)、阳性对照药戊酸雌二醇片(estradiol valerate tablets,EVT)和蒸馏水给大鼠灌胃,各组灌胃剂量分别为18.9,20.52 g·kg-1,0.36 mg·kg-1,1.0 mL·kg-1,日2次,于第4天给药2 h后,腹主动脉取血,离心,56℃水浴灭活30 min后,大鼠含药血清制备完成。再配制10%各组含药血清的α-MEM分别加入诱导剂(YDJ)(10-7mol·L-1地塞米松、50μmol·L-1维生素C、10 mmol·L-1β-甘油磷酸钠)与单纯诱导剂组共5组对BMSCs(1×105/L)进行干预9 d,用Westernblot法检测碱性磷酸酶(alkaline phosphatase,ALP)、Ⅰ型胶原(typeⅠcollagen,COLⅠ)蛋白表达,用RT-PCR法检测转化生长因子β1(transforming growth factor beta 1,TGF-β1)mRNA表达。结果:Control组比较:ZGW+YDJ组、YGW+YDJ组、EVT+YDJ组均可促进BMSCs的ALP,COLⅠ蛋白的表达,上调BMSCs TGF-β1mRNA表达(P<0.05);与YDJ组比较:ZGW+YDJ组、YGW+YDJ组可促进BMSCs的ALP,COLⅠ蛋白的表达,上调BMSCs TGF-β1mRNA表达(P<0.05);且ZGW+YDJ组优于YGW+YDJ组(P<0.05)。结论:左、右归丸含药血清能协同诱导剂促进BMSCs成骨分化,且左归丸组更为有效,表明中医"滋肾阴"对BMSCs的成骨诱导更为有效。Objective： To observe the effects of serum containing Zuogui Wan and Yougui Wan on the osteogenic differentiation in bone marrow mesenchymal stem cells（BMSCs）.Method： To assign 28 rats into 4 groups at random,then the rats were respectedly gavaged with Zuogui Wan,Yougui Wan,positive control drug（estradiol valerate,EVT） and distilled water.The dosage of gavage was respectively 18.9,20.52 g·kg-1,0.36 mg·kg-1,1.0 mL·kg-1,two times one day.On the 4th day,after being gavaged 2 h,get the containing serum from aorta abdominalis,centrifuge,inactivate（56 ℃,30 min）.The four kinds of 10% containing serum separately adding inducer（10-7mol ·L-1dexamethasone,50 μmol ·L-1vitamin C,10 mmol ·L-1sodium β-glycerophosphate）,with inducer group,simultaneously intervene the BMSCs（1 × 105/ L） for about 9 days.Then,to show the expression of alkaline phosphatase（ALP） and type Ⅰ collagen（COL Ⅰ） protein with the method of Western blot,to test expressing transforming growth factor beta 1（TGF-β 1） mRNA by RT-PCR.Result： Compared with control group,Zuigui Wan,Yougui Wan,EVT can promote BMSCs ALP,COL Ⅰ expression,upregulate BMSCs TGF-β 1 mRNA（P 0.05）;compared with inducer group,Zuigui Wan,Yougui Wan can promote BMSCs ALP,COLⅠexpression,upregulate BMSCs TGF-β 1 mRNA,also Zuigui Wan is better than Yougui Wan（P 0.05）.Conclusion： Serum containing Zuigui Wan and Yougui Wan can help inducer on the osteogenic differentiation in BMSCs,and Zuigui Wan is better,which indicates nourishing kidney-Yin is more effective on osteogenic differentiation in BMSCs.

关 键 词：左归丸 右归丸 骨髓间充质干细胞 碱性磷酸酶 Ⅰ型胶原 转化生长因子Β1 

分 类 号：R285.5[医药卫生—中药学]