复方补肾活血颗粒含药血清通过Trb3调控hBMSCs成骨成脂分化  

Compound Kidney-Invigorating Granule medicated serum regulates osteogenic and adipogenic differentiation of human bone marrow mesenchymal stem cells via Trb3

作　　者：王子文 王久香 吕浩 胡芷苜 江渟 WANG Ziwen;WANG Jiuxiang;LV Hao;HU Zhimu;JIANG Ting(Graduate School,Anhui University of Chinese Medicine,Hefei 230031,China;Experimental Center of Clinical Research,The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学研究生院,安徽合肥230031 [2]安徽中医药大学第一附属医院临床试验中心,安徽合肥230031

出　　处：《中国骨质疏松杂志》2023年第4期544-549,561,共7页Chinese Journal of Osteoporosis

基　　金：安徽省重点研究与开发计划(202104j07020010);安徽中医药大学第一附属医院临床科学研究项目(2020yfyzc27);新安医学教育部重点实验室开放项目(2022XAYX12)。

摘　　要：目的研究复方补肾活血颗粒含药血清通过Trb3调节人骨髓间充质干细胞(human bone mesenchymal stem cells,hBMSCs)成骨/成脂分化。方法不同浓度复方补肾活血颗粒含药血清干预hBMSCs,通过CCK8法检测细胞活力,ALP活性、茜素红染色观察hBMSCs成骨分化,油红O染色鉴定脂肪分化。Western blot检测成骨/成脂标志物蛋白表达。Western blot和qPCR检测Trb3蛋白和基因表达。Trb3 siRNA转染hBMSCs后通过Western blot观察成骨成脂相关因子蛋白表达情况。结果复方补肾活血颗粒含药血清以浓度依赖性促进hBMSCs增殖。复方补肾活血颗粒含药血清增强hBMSCs中ALP活性及矿化结节,上调Runx2、Osterix蛋白表达并抑制PPARγ、FABP4蛋白表达和脂质积累。机制研究发现,复方补肾活血颗粒含药血清促进Trb3表达,当内源性Trb3敲低时,逆转了复方补肾活血颗粒含药血清促进成骨分化抑制脂肪分化的作用。结论复方补肾活血颗粒含药血清通过Trb3以牺牲脂肪分化为代价,促进hBMSCs成骨分化。Objective To investigate the regulation of osteogenic/adipogenic differentiation of human bone marrow mesenchymal stem cells via Trb3 through Compound Kidney-Invigorating Granules(CKG)medicated serum.Methods hBMSCs were intervened with different concentrations of CKG-medicated serum.Cell viability was measured by CCK8 assay,hBMSCs osteogenic differentitation was observed by ALP activity and alizarin red staining,and adipogenic differentiation by oil red O staining.hBMSCs were examined by Western blot for osteogenic/adipogenic marker protein expression.Western blot and qPCR were performed to defect Trb3 protein and gene expression.Trb3 siRNA was transfected with hBMSCs to observe the expression of osteogenic/adipogenic related factors by Western blot.Results CKG medicated serum promoted hBMSCs proliferation in a concentration-dependent manner.CKG medicated serum enhanced ALP activity and mineralized nodules.It upregulated Runx2 and Osterix protein expression,and inhibited PPARγand FABP4 protein expression and lipid accumulation.Mechanistic studies revealed that Trb3 knockdown reversed the ability of CKG medicated serum to promote osteogenic differentiation and inhibit adipogenic differentiation.Conclusion CKG-medicated serum promotes osteogenic differentiation of hBMSCs via Trb3 at the expense of adipogenic differentiation.

关 键 词：骨质疏松症 复方补肾活血颗粒 TRB3 成骨分化 成脂分化 

分 类 号：R285.5[医药卫生—中药学]