整合素α2在老龄骨质疏松BMSC成骨分化的调控机制  

Mechanisms of integrin alpha2 in osteogenic differentiation of BMSC in aged osteoporosis

作　　者：蒋代富 朱晓英 肖雪 JIANG Dai-Fu;ZHU Xiao-Ying;XIAO Xue(Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou, China)

机构地区：[1]遵义医科大学附属医院

出　　处：《中国老年学杂志》2019年第20期5055-5058,共4页Chinese Journal of Gerontology

基　　金：国家“十二五”科技支撑计划课题(2012BAI10B01)

摘　　要：目的 分析整合素α2在老龄骨质疏松骨髓间充质干细胞(BMSCs)成骨分化的调控机制。方法 检测慢病毒转染BMSCs效率、骨质疏松患者BMSCs中整合素α2转染后高表达,分析骨质疏松患者BMSCs成骨、增殖受到整合素α2高表达的影响及作用机制。结果 BMSCs对整合素α2进行转染3d后,荧光显微镜下可见有GFP表达,感染复数(MOI)=10、20、50、100时视野中分别有零星荧光、少许荧光、显著较多的荧光、最多的荧光。转染后BMSC整合素α2表达显著高于野生型(P<0.05)。整合素α2蛋白表达在慢病毒转染后显著提升,免疫荧光强度显著增强。培养4d后,细胞向增殖期进入,具有较大的生长曲线斜度,说明细胞增殖速度在转染后明显加快,骨质疏松患者BMSC整合素α高表达能够为细胞增殖提供良好的前提条件。对比转染后野生型Osterix、RUNX2表达显著提升。对整合素α2进行转染能够促进骨质疏松BMSC矿化能力的增强。骨质疏松患者BMSC整合素α高表达能够为成骨分化提供良好的前提条件。成骨诱导后30min,转染后细胞外信号调节激酶(ERK)快速活化,1h时显著低于未转染组(P<0.05),但是二者的蛋白激酶B(AKT)/JNK/P38之间的差异不显著(P>0.05)。整合素将ERK1/2通路活化,从而为细胞增殖、分化提供了良好的前提条件。PD98059能够抑制活化ERK、转染后RUNX2,进而抑制茜素红染色所体现的成骨分化、碱性磷酸酶(ALP)活性。结论 整合素α2/ERK/Runx2信号通路参与了老龄骨质疏松发病,为老龄骨质疏松的预防与治疗提供了新的分子机制。Objective To analyze the regulation mechanism of integrin alpha2 on osteogenic differentiation of BMSC in aged osteoporosis. Methods The efficiency of lentivirus transfection of BMSC and the high expression of integrin alpha2 in BMSC of osteoporosis patients were detected. The mechanism of BMSC osteogenesis and proliferation in osteoporosis patients affected by the high expression of integrin alpha2 and the osteogenesis and proliferation of BMSC in osteoporosis patients affected by the high expression of integrin alpha2 were analyzed. Results Three days after transfection of integrin alpha2 by MSCs, GFP expression was observed under fluorescence microscope. Sporadic fluorescence, significant fluorescence and maximum fluorescence were observed in the field of vision at MOI=10, 20, 50 and 100. The positive rate of GFP after transfection was found under fluorescence microscope to be MOI=50, above 90% at 100, and the optimal MOI=50. The expression of BMSC integrin alpha2 after transfection was significantly higher than that of wild type BMSC ( P <0.05). The expression of integrin alpha2 protein was increased significantly after lentivirus transfection, and the immunofluorescence intensity was increased significantly. After 4 days of culture, the cells entered the proliferative phase with a large slope of growth curve, indicating that the cell proliferation rate was significantly accelerated after transfection. The high expression of BMSC integrin alpha in osteoporosis patients provided a good prerequisite for cell proliferation. The expression of wild-type Osterix and RUNX2 were increased significantly after transfection, and the activities of ALP were increased by 43.6%, 58.0% and 60.4% at the 3rd, 1st and 2nd week, respectively. Transfection of integrin alpha2 promoted the mineralization of osteoporotic BMSCD, which were increased by 40.8% and 54.8% in 2 and 3 weeks respectively. The high expression of BMSC integrin alpha in osteoporotic patients provided a good prerequisite for osteogenic differentiation. At 30 mi

关 键 词：整合素Α2 骨质疏松 骨髓间充质干细胞 

分 类 号：R580[医药卫生—内分泌]