补肾活血胶囊介导Hedgehog信号通路调控大鼠骨髓间充质干细胞成骨成脂分化  

Bushen Huoxue capsule regulates osteogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells via Hedgehog signaling pathway

作　　者：梁学振 杨曦[2] 李嘉程 骆帝 许波 李刚 Liang Xuezhen;Yang Xi;Li Jiacheng;Luo Di;Xu Bo;Li Gang(First Clinical Medical College of Shandong University of Traditional Chinese Medicine,Jinan 250355,Shandong Province,China;Laboratory Department of Xinjiang Military Region General Hospital,Urumqi 830000,Xinjiang Uygur Autonomous Region,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,Shandong Province,China)

机构地区：[1]山东中医药大学第一临床医学院,山东省济南市250355 [2]新疆军区总医院检验科,新疆维吾尔自治区乌鲁木齐市830000 [3]山东中医药大学附属医院,山东省济南市250014

出　　处：《中国组织工程研究》2022年第7期1020-1026,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81774333),项目负责人:李刚。

摘　　要：背景:激素性股骨头坏死是骨科常见的进行性难治性疾病,严重影响患者的正常生活和工作。骨髓间充质干细胞成骨成脂分化异常被证实参与了激素性股骨头坏死的发生发展及防治,但其作用机制尚不清楚。目的:观察补肾活血胶囊对糖皮质激素处理的骨髓间充质干细胞成骨-成脂分化及Hedgehog信号通路相关因子的影响。方法:分离培养SD大鼠骨髓间充质干细胞,运用不同剂量的补肾活血胶囊含药血清及糖皮质激素干预,CCK-8法检测细胞增殖活性,茜素红染色和油红O染色观察骨髓间充质干细胞成骨和成脂分化情况,Western blot检测细胞中成骨、成脂及Hedgehog信号通路相关因子的蛋白表达变化。结果与结论:①补肾活血胶囊含药血清不仅能促进正常大鼠骨髓间充质干细胞的增殖,而且能明显改善糖皮质激素对骨髓间充质干细胞增殖的抑制效应;②成骨诱导3周后,补肾活血胶囊含药血清组与空白对照组相比成骨诱导能力增强,尤其是补肾活血胶囊中剂量组表现出更高的成骨活性;③成脂诱导2周后,油红O染色显示补肾活血胶囊含药血清组与空白对照组相比阳性染色面积均明显减少,其中补肾活血胶囊中剂量组抑制程度最为明显;④各组干预3 d后,Western blot检测显示补肾活血胶囊含药血清均能增强骨髓间充质干细胞中成骨因子Runx2、Col1a1和Hedgehog信号通路相关因子Shh、Gli1的蛋白表达,减弱成脂因子PPARγ、C/EBPα的蛋白表达;⑤结果表明,补肾活血胶囊含药血清能介导Hedgehog信号通路促进骨髓间充质干细胞成骨分化、抑制其成脂分化。BACKGROUND:Steroid-induced osteonecrosis of femoral head,a common,progressive and refractory orthopedics disease,seriously affected the normal life and work of the patients.The imbalance of osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was confirmed to involve in the pathogenesis,prevention,and treatment of steroid-induced osteonecrosis of femoral head.However,the mechanism is unclear.OBJECTIVE:To observe the effect of Bushen Huoxue capsule containing serum on osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells treated with glucocorticoids and Hedgehog signal pathway related factors.METHODS:Bone marrow mesenchymal stem cells were isolated from SD rats and cultured,and intervened with different doses of Bushen Huoxue capsule containing serum and glucocorticoids.CCK-8 assay was used to observe the proliferation of cells.The osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells in different groups were detected by alizarin red staining and oil red“O”staining.The changes of protein expression of osteogenic factors and adipogenic factors and Hedgehog signal pathway related factors of bone marrow mesenchymal stem cells in different groups were analyzed by western blot assay.RESULTS AND CONCLUSION:(1)The Bushen Huoxue capsule containing serum not only could promote the proliferation of normal rat bone marrow mesenchymal stem cells,but also could obviously increase the proliferation activity of bone marrow mesenchymal stem cells treated by the glucocorticoids.(2)After 3 weeks of bone induction,osteogenic induction abilities in the low dose group,middle dose group and high dose group of Bushen Huoxue capsule containing serum were significantly higher than that of the blank control group,especially the middle dose group.(3)After 2 weeks of lipid induction,the positive staining areas revealed by oil red“O”staining in the low dose group,middle dose group and high dose group of Bushen Huoxue capsule containing serum were lower than

关 键 词：干细胞 骨髓间充质干细胞 激素性股骨头坏死 补肾活血胶囊 HEDGEHOG信号通路 成骨分化 成脂分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]