Klotho对大鼠骨髓间充质干细胞增殖和分化的影响

doi:10.16098/j.issn.0529-1356.2019.06.005

解剖学报 ›› 2019, Vol. 50 ›› Issue (6): 729-734.doi: 10.16098/j.issn.0529-1356.2019.06.005

Klotho对大鼠骨髓间充质干细胞增殖和分化的影响

李秀全¹柏树令¹敖强¹王小红¹ 常世杰² 田晓红¹佟浩¹ 范军^1*

1.中国医科大学公共基础学院组织工程学教研室，沈阳 110001;2.中国医科大学公共基础学院生物医学工程学教研室，沈阳 110001

收稿日期:2018-11-07 修回日期:2019-01-02 出版日期:2019-12-06 发布日期:2019-12-06
通讯作者: 范军 E-mail:cmu_fanjun@163.com
基金资助:
国家自然科学基金

Effects of Klotho on the proliferation and differentiation of rat bone marrow mesenchymal stem cells

LI Xiu-quan¹BAI Shu-ling¹AO Qiang¹WANG Xiao-hong¹CHANG Shi-jie² TIAN Xiao-hong¹ TONG Hao¹ FAN Jun ^1*

1.Department of Tissue Engineering, College of Fundamental Sciences, China Medical University, Shenyang 110001，China; 2.Department of Biomedical Engineering, College of Fundamental Sciences, China Medical University, Shenyang 110001，China

Received:2018-11-07 Revised:2019-01-02 Online:2019-12-06 Published:2019-12-06
Contact: FAN Jun E-mail:cmu_fanjun@163.com

摘要/Abstract

摘要：

目的探讨延缓衰老基因Klotho对骨髓间充质干细胞（BMSCs）增殖和分化的影响。方法体外条件下培养大鼠骨髓间充质干细胞，构建分泌型Klotho(sKL)过表达的BMSCs。Western blotting检测细胞及培养基中sKL的表达；MTT法检测细胞增殖能力；Western blotting检测衰老标志物P53、P21蛋白的表达。利用成骨诱导液定向诱导BMSCs向成骨细胞分化，应用碱性磷酸酶(ALP)染色鉴定成骨效果；利用成脂诱导液定向诱导BMSCs向脂肪细胞分化，应用油红O染色鉴定成脂效果。结果 Western blotting结果显示，sKL组细胞和培养基中sKL蛋白表达显著上调（P<0.05），P53、P21表达显著下调（P<0.05）；MTT结果显示，各组细胞吸光度值（A值）差别无显著性（P>0.05），sKL组成骨及成脂能力明显强于对照组（P<0.05）。结论 sKL增强了大鼠BMSCs的延缓衰老能力，对BMSCs的成骨分化和成脂分化产生一定的促进作用，而对BMSCs的增殖无显著影响。

关键词: 骨髓间充质干细胞, 分泌型Klotho, 增殖, 分化, 免疫印迹法

Abstract:

Objective To investigate the effect of antiaging gene Klotho on the proliferation and differentiation of rat bone marrow mesenchymal stem cells(BMSCs). Methods Rat BMSCs overexpressing secreted Klotho(sKL) were cultured and constructed in vitro. MTT assay was used to detect the cell proliferation. Western blotting was used to measure the expression of sKL in the cells and medium of three groups. As markers of aging, P53 and P21 in three cells were also detected. BMSCs were induced to osteogenic differentiation by osteogenic induction medium, the result of differentiation was observed by alkaline phosphatase(ALP) staining; BMSCs were induced to adipogenic differentiation by adipogenic induction medium, the result of differentiation was identified by oil red O staining. Results Western blotting showed that the expression of sKL increased significantly in the sKL group (P<0.05), in addition, the expression of P53 and P21 decreased significantly in the sKL group(P<0.05). MTT assay indicated that there was no significant differences about absorbance value between sKL group and control group(P>0.05). The ability of osteogenic differentiation and adipogenic differentiation was significantly stronger than that of the control group(P<0.05). Conclusion sKL effectively enhances the anti-aging ability of BMSCs, meanwhile it also promotes osteogenic and adipogenic differentiation of BMSCs. Besides, this study also suggests that Klotho may have no effect on the proliferation of BMSCs.

Key words: Bone marrow mesenchymal stem cell, Secreted Klotho, Proliferation, Differentiation, Western blotting

李秀全柏树令敖强王小红常世杰田晓红佟浩范军. Klotho对大鼠骨髓间充质干细胞增殖和分化的影响[J]. 解剖学报, 2019, 50(6): 729-734.

LI Xiu-quan BAI Shu-ling AO Qiang WANG Xiao-hong CHANG Shi-jie TIAN Xiao-hong TONG Hao FAN Jun. Effects of Klotho on the proliferation and differentiation of rat bone marrow mesenchymal stem cells[J]. Acta Anatomica Sinica, 2019, 50(6): 729-734.

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Klotho对大鼠骨髓间充质干细胞增殖和分化的影响

Effects of Klotho on the proliferation and differentiation of rat bone marrow mesenchymal stem cells

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