补肾活血汤对地塞米松处理后大鼠骨髓间充质干细胞的影响  

Effect of Bushen Huoxue Decoction on Bone Marrow Mesenchymal Stem Cells in Rats Treated with Dexamethasone

作　　者：刘浩 骆帝 阎伟[2] 李金松[2] 闫德志 LIU Hao;LUO Di;YAN Wei;LI Jinsong;YAN Dezhi(The First Clinical Medical College,Shandong University of Traditional Chinese Medicine,Ji'nan 250013,China;Orthopedics Department,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Ji'nan 250013,China)

机构地区：[1]山东中医药大学第一临床医学院,济南250013 [2]山东中医药大学附属医院关节骨科,济南250013

出　　处：《世界中医药》2025年第3期411-418,423,共9页World Chinese Medicine

基　　金：国家自然科学基金项目(82074453,82205154);山东省重点研发计划(重大科技创新工程)项目(2021CXGC010501);山东省自然科学基金青年项目(ZR2021QH004);山东省自然科学基金联合专项基金项目(ZR2021LZY002);2020年齐鲁卫生与健康杰青人才培育工程项目(rc2021002-23)。

摘　　要：目的:基于刺猬(Hh)信号通路探讨补肾活血汤含药血清对地塞米松(Dex)处理后大鼠骨髓间充质干细胞(BMSC)增殖、迁移及成骨分化的影响。方法:贴壁法提取大鼠BMSC,评估纯度和活性。将大鼠BMSC分为8组:对照组、补肾活血汤低、中、高剂量组(0.662、1.323、2.646 g/kg补肾活血汤灌胃大鼠血清)、Dex组、Dex+补肾活血汤低、中、高剂量组(Dex+0.662、1.323、2.646 g/kg补肾活血汤灌胃大鼠血清)。干预48 h后,细胞计数试剂盒-8(CCK-8)法检测BMSC增殖,细胞划痕实验检测BMSC迁移,荧光定量聚合酶链反应(qPCR)检测BMSC中成骨相关指标[碱性磷酸酶(ALP)、Ⅰ型胶原蛋白(COLⅠ)、Runt相关转录因子2(RUNX2)、骨桥蛋白(OPN)]及Hh信号通路因子[音猬因子(Shh)、神经胶质瘤相关癌基因同源蛋白1(Gli1)、Gli2]mRNA表达。结果:贴壁法提取的BMSC具有较高纯度和良好的分化潜能,补肾活血汤含药血清能显著逆转Dex对BMSC增殖、迁移和成骨分化潜能的抑制作用。与Dex组比较,中剂量补肾活血汤含药血清上调BMSC中成骨因子(ALP、COL I、RUNX2)和Hh信号通路因子(Gli1、Gli2、Shh)mRNA和蛋白表达(均P<0.05),中、高剂量补肾活血汤含药血清上调OPN蛋白表达(均P<0.05)。结论:补肾活血汤含药血清能通过上调Hh信号通路因子,促进Dex处理后BMSC增殖、迁移及成骨分化。Objective:To investigate the effect of Bushen Huoxue Decoction(BHD)-medicated serum on the proliferation,migration,and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)treated with dexamethasone(Dex)via the Hedgehog(Hh)signaling pathway.Methods:BMSCs were isolated from rats using the adherent culture method,and their purity and viability were assessed.The BMSCs were divided into eight groups:control,low-,medium-,and high-dose BHD groups(serum from rats receiving 0.662,1.323,and 2.646 g/kg BHD,respectively),Dex group,and Dex+low-,medium-,and high-dose BHD groups(Dex+serum from rats receiving 0.662,1.323,and 2.646 g/kg BHD,respectively).After 48 hours of intervention,cell proliferation was assessed using the Cell Counting Kit-8(CCK-8),and migration was evaluated using a scratch assay.The mRNA expression of osteogenic differentiation markers[alkaline phosphatase(ALP),type I collagen(COL I),Runt-related transcription factor 2(RUNX2),and osteopontin(OPN)]and Hh signaling pathway factors[Sonic hedgehog(Shh),glioma-associated oncogene homolog 1(Gli1),and Gli2]was analyzed using quantitative real-time PCR(qPCR).Results:BMSCs isolated by the adherent culture method exhibited high purity and good differentiation potential.BHD-medicated serum significantly reversed the inhibitory effects of Dex on BMSC proliferation,migration,and osteogenic differentiation.Compared with the Dex group,the medium-dose BHD-medicated serum significantly upregulated the mRNA and protein expression of osteogenic markers(ALP,COL I,RUNX2)and Hh signaling pathway factors(Shh,Gli1,Gli2)in BMSCs(all P<0.05).Moreover,both medium-and high-dose BHD-medicated serum significantly increased OPN protein expression(both P<0.05).Conclusion:BHD-medicated serum promotes the proliferation,migration,and osteogenic differentiation of Dex-treated BMSCs by upregulating Hh signaling pathway factors.

关 键 词：补肾活血汤 含药血清 骨髓间充质干细胞 地塞米松 刺猬信号通路 细胞增殖 细胞迁移 细胞成骨分化 

分 类 号：R282[医药卫生—中药学]