不同形式运动对胰岛素抵抗小鼠骨髓间充质干细胞向成骨细胞分化能力的影响研究

Effect of different kinds of exercise training on bone marrow mesenchymal stem cells to osteoblast differentiation ability in mice of insulin resistance

目的探讨在T2DM的发生发展中胰岛素和运动对成骨细胞的影响。方法选取40只6周龄雄性C57BL/6小鼠为研究对象,按体重分为正常对照组(NC,n=10)和高脂组(HF,n=30),高脂饲料喂养12周构建IR模型。建模成功后,HF组按体重分为IR组(n=9)、IR+跑台组(IR+RT,n=10)和IR+爬梯组(IR+LS,n=10),随后进行为期12周的运动干预。提取小鼠右腿胫骨骨髓间充质干细胞(BMSC)进行体外培养,菌落形成单位(CFU)法检测其增殖情况;RT-PCR检测BMSC中碱性磷酸酶(ALP)、锌指结构转录因子(Osterix)、激活转录因子4(ATF4)、骨钙素(OCN)、骨桥蛋白(OPN)、Runt相关转录因子2(Runx2)、CCAAT/增强子结合蛋白(CEBP)和核因子κB受体活化因子配体(RANKL)mRNA的表达。结果 CFU实验结果显示,IR+RT组六孔板的成骨细胞数目最多。BMSC中基因检测发现,与NC组比较,IR+RT组ALP、Osterix、OCN、OPN mRNA表达上调(P<0.05),IR+LS组Osterix和OCN mRNA(P<0.05)、OPN mRNA(P<0.01)表达上调;与IR组比较,IR+RT组和IR+LS组OCN mRNA表达上调(P<0.05);与IR+RT组比较,IR+LS组ATF4 mRNA表达上调(P<0.05)。结论有氧运动和负重运动均可改善IR小鼠BMSC向成骨细胞分化的能力,在分化早期阶段有氧运动的影响更显著。

Objective To explore the effect Of insulin and exercise on osteoblast in the development of type 2 diabetes （T2DM）. Methods 40 six-week-old C57BL/6 mate mice were divided into control group （ NC, n -- 10 ） and high-fat diet group （HF, n = 30 ） in accordance with weight. We established insulin resistance mice by high-fat diet. The model mice were divided into insulin resistance group （IR, n= 9）, insulin resistance ＋ running treadmill group （IR＋RT, n= 10）, and insulin resistance ~ ladder stand group （IR＋LS,n=10）. After 12 weeks exercise,we extracted mice＇s right-tibia BMSC and tested their reproduction by CFU. RNA in BMSC and the expression levels of ALP, Osterix, ATF4, OCN, OPN, Runx2,CEBP and RANKL mRNA were evaluated by RT-PCR. Results CFU data showed that the numbers of osteoblasts in the six-well plates of IR＋RT group were the most. Compared with NC group, the expressions of ALP, Osterix, OCN and OPN mRNA were up-regulated in IR＋RT group （P〈0. 05） ; the expressions of Osterix,OCN mRNA （P〈0. 05） and OPN mRNA （P〈0.01） were up-regulated in IR＋LS group. Compared with IR group, the expressions of OCN mRNA was up-regulated in IR＋RT group and IR＋LS group （P〈0. 05）. Compared with IR＋RT group,the expressions of ATF4 mRNA was up- regulated in IR＋LS group （P〈0. 05）. Conclusion Aerobic exercise and weight-bearing exercise can improve the ability of BMSC＇s differentiation into osteoblast in mice with IR. In the early process of BMSC differentiation into osteoblasts, aerobic exercise play a more important role.