辛伐他汀可刺激骨髓间充质干细胞的成骨分化  

Simvastatin stimulates osteogenic differentiation of bone marrow mesenchymal stem cells

作　　者：郭志斌 吴春芳 刘子洪[2] 张钰英 迟博婧 王宝 马超[2] 张国彬 田发明 Guo Zhibin;Wu Chunfang;Liu Zihong;Zhang Yuying;Chi Bojing;Wang Bao;Ma Chao;Zhang Guobin;Tian Faming(Department of Orthopedic Surgery,Kailuan Linxi Hospital,Tangshan 063000,Hebei Province,China;Department of Orthopedic Surgery,Kailuan Hospital,Tangshan 063000,Hebei Province,China;Medical Research Center,North China University of Science and Technology,Tangshan 063000,Hebei Province,China)

机构地区：[1]开滦总医院林西医院骨外科,河北省唐山市063000 [2]开滦总医院骨外科,河北省唐山市063000 [3]华北理工大学医学实验研究中心,河北省唐山市063000

出　　处：《中国组织工程研究》2021年第19期2963-2968,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81874029),项目负责人:田发明;河北省医学科学研究重点课题(20190152),项目负责人:郭志斌;河北省医学科学研究重点课题(20160722),项目负责人:张国彬;河北省自然科学基金(H2013209255),项目负责人:田发明。

摘　　要：背景:辛伐他汀可显著刺激骨髓间充质干细胞成骨分化,但机制不明。近年研究证实p38 MAPK信号通路参与调控骨髓间充质干细胞向成骨细胞分化的过程。目的:观察p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路在辛伐他汀体外干预刺激骨髓间充质干细胞成骨分化中的作用。方法:分离培养SD大鼠股骨和胫骨骨髓间充质干细胞,第2代骨髓间充质干细胞分为3组:对照组、辛伐他汀组(加入10^-7 mol/L辛伐他汀)、阻断剂组(加入辛伐他汀30 min前给予10μmol/L p38MAPK信号通路特异性阻断剂SB203580),均使用含有10 mmol/Lβ-甘油酸磷酸钠、50 mg/L抗坏血酸的DMEM完全培养基进行成骨诱导分化。各组干预第6天进行碱性磷酸酶染色;第6天和第12天采用Western blot法检测p38 MAPK和磷酸化p38 MAPK的表达;第12天采用免疫荧光染色和实时荧光定量聚合酶链反应检测骨钙素、Ⅰ型胶原的表达;第21天茜素红染色观察钙结节形成情况。结果与结论:①辛伐他汀组碱性磷酸酶表达和基质矿化能力显著高于对照组,阻断剂组显著低于辛伐他汀组(P<0.05);②辛伐他汀组磷酸化p38 MAPK与p38 MAPK比值高于对照组(P<0.05),阻断剂组低于辛伐他汀组(P<0.05);③辛伐他汀可以促进骨钙素和Ⅰ型胶原的表达,而阻断剂组显著低于辛伐他汀组(P<0.05);④结果表明,辛伐他汀可以促进骨髓间充质干细胞向成骨细胞分化,这一作用可能与促进p38 MAPK磷酸化进而提高该通路活性有关。BACKGROUND: Simvastatin can remarkably stimulate the osteogenic differentiation of bone marrow mesenchymal stem cells, but the mechanism is unknown. Recent studies have confirmed that p38 mitogen-activated protein kinase(MAPK) signaling pathway is involved in regulating the differentiation of bone marrow mesenchymal stem cells into osteoblasts. OBJECTIVE: To observe the role of p38 MAPK signaling pathway in simvastatin stimulated osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells from the femur and tibia of Sprague-Dawley rats were cultured in vitro, and the second generation of bone marrow mesenchymal stem cells was divided into three groups: control group, simvastatin group(10-7 mol/L simvastatin), and blocking agent group(10 μmol/L p38 MAPK signaling pathway specific blocker SB203580 30 minutes before adding simvastatin). Osteogenic differentiation was induced in DMEM complete medium containing 10 mmol/L β-glycerophosphate and 50 mg/L ascorbic acid. Alkaline phosphatase staining was performed at 6 days after intervention in each group. The expressions of p38 MAPK and phosphorylated p38 MAPK were detected by western blot assay at 6 and 12 days. The expression of osteocalcin and collagen type Ⅰ was detected by immunofluorescence and real-time fluorescent quantitative polymerase chain reaction at 12 days. The formation of calcium nodules was observed by alizarin red staining at 21 days. RESULTS AND CONCLUSION:(1) Alkaline phosphatase expression and matrix mineralization ability were significantly higher in the simvastatin group than those of control group, and significantly lower in the blocker group than those in the simvastatin group(P < 0.05).(2) The ratio of p38 MAPK in simvastatin group was significantly higher than that in control group(P < 0.05), and that in blocker group was significantly lower than that in simvastatin group(P < 0.05).(3) Simvastatin could promote the expression levels of osteocalcin and collagen type Ⅰ, while above expressi

关 键 词：干细胞 骨髓间充质干细胞 辛伐他汀 P38 MAPK 信号通路 成骨分化 成骨细胞 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]