基于lncRNA H19/miR-22/Wnt通路探索补肾强督方干预人骨髓间充质干细胞成骨分化的作用机制  

Exploration on the mechanism of Bushen Qiangdu Formula intervention on osteogenic differentiation of human bone marrow mesenchymal stem cells based on the lncRNA H19/miR-22/Wnt pathway

作　　者：孙文婷 吴娟 穆闻君 郑丹妮 夏启胜[2] 陈志华[2] 周童亮[2] 刘婷[1] 陶庆文[1,3] 阎小萍 孔维萍[1,3] SUN Wenting;WU Juan;MU Wenjun;ZHENG Danni;XIA Qisheng;CHEN Zhihua;ZHOU Tongliang;LIU Ting;TAO Qingwen;YAN Xiaoping;KONG Weiping(Department of Rheumatology,China-Japan Friendship Hospital,Beijing 100029,China;Institute of Clinical Medicine,China-Japan Friendship Hospital,Beijing 100029,China;Key Laboratory for Immune Inflammatory Disease of Beijing,Beijing 100029,China)

机构地区：[1]中日友好医院中医风湿病科,北京100029 [2]中日友好医院临床研究所,北京100029 [3]免疫炎性疾病北京市重点实验室,北京100029

出　　处：《中华中医药杂志》2025年第1期145-150,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82374369);中央高水平医院临床科研业务费资助(No.2022-NHLHCRF-LX-02-0104);中日友好医院临床研究与转化跃升项目(No.2023-NHLHCRF-YYPPLC-TJ-02);首都卫生发展科研专项(No.2024-2-40613)。

摘　　要：目的:研究补肾强督方对人骨髓间充质干细胞(hBMSCs)成骨分化的影响及其作用机制。方法:用补肾强督方含药血清处理hBMSCs,诱导成骨分化14 d,茜素红染色评估矿化程度,RT-PCR检测成骨细胞标志物及H19、miR-22的表达,Western Blot检测β-catenin和DKK-1的表达。构建H19过表达的hBMSCs,并加入补肾强督方高剂量含药血清干预,诱导成骨分化14 d后再次检测上述指标。结果:与空白对照组比较,补肾强督方含药血清抑制hBMSCs成骨分化,补肾强督方高剂量组可降低Wnt通路中的β-catenin表达(P<0.01),上调DKK-1的表达(P<0.01),并降低H19的表达(P<0.05),增加miR-22的表达(P<0.01)。与H19过表达组比较,补肾强督方高剂量含药血清能部分挽救H19过表达引起的hBMSCs成骨分化(P<0.01,P<0.05),调节DKK-1和β-catenin的表达(P<0.01,P<0.05)。结论:补肾强督方可能通过lncRNA H19/miR-22/Wnt通路干预hBMSCs的成骨分化,为该方在AS病理性成骨中的应用提供了理论依据。Objective:To investigate the effects and mechanisms of Bushen Qiangdu Formula(BSQD) on the osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs).Methods:hBMSCs were treated with BSQDcontaining serum to induce osteogenic differentiation for 14 d.Alizarin Red staining was performed to evaluate the degree of mineralization.RT-PCR was used to detect osteogenic cell markers,as well as the expression of H19 and miR-22.Western Blot was conducted to examine the expression of β-catenin and DKK-1.hBMSCs with overexpression of H19 were constructed,and high-dose BSQD-containing serum was added to intervene.After inducing osteogenic differentiation for 14 d,the abovementioned indicators were re-evaluated.Results:Compared with control group,BSQD-containing serum could inhibit the osteogenic differentiation of hBMSC,high-dose BSQD-containing serum could reduce the expression of β-catenin in the Wnt pathway(P<0.01),upregulate the expression of DDK-1(P<0.01),and decrease the expression of H19(P<0.05) while increasing the expression of miR-22(P<0.01).Compared with H19 overexpression group,high-dose BSQD-containing serum could partially rescue the osteogenesis caused by H19 overexpression in hBMSCs(P<0.01,P<0.05),regulate the expression of DKK-1 and β-catenin(P<0.01,P<0.05).Conclusion:BSQD may intervene in the bone formation and differentiation of hBMSCs through the lncRNA H19/miR-22/Wnt pathway,providing a theoretical basis for the application of BSQD in pathological osteogenesis.

关 键 词：补肾强督方 人骨髓间充质干细胞 成骨分化 强直性脊柱炎 

分 类 号：R285[医药卫生—中药学]