整合素β_(1)在大鼠BMSCs软骨分化过程中的实验研究  

Experimental Study of Integrinβ_(1)in the Process of Cartilage Differentiation in Rat BMSCs

作　　者：邓娟 左右清 王念[2] 胡熙苒 曾礼 Deng Juan;Zuo Youqing;Wang Nian(Clinical School,Xiangtan Medical and Health Vocational and Technical College,Xiangtan,Hunan,411104,China)

机构地区：[1]湘潭医卫职业技术学院临床学院,湖南湘潭411104 [2]中南大学湘雅医学院基础医学院,湖南长沙410013

出　　处：《黑龙江医学》2023年第17期2080-2082,2087,共4页Heilongjiang Medical Journal

基　　金：湘潭医卫职业技术学院2021年度重点自科项目(ZRKX2021-04)。

摘　　要：目的:研究和探讨整合素β_(1)在转化生长因子(TGF)诱导BMSCs分化为软骨细胞过程中的作用。方法:全骨髓法体外培养大鼠骨髓基质干细胞(BMSCs),TGF-β3促进和诱导BMSCs的软骨分化、抗整合素β_(1)单克隆抗体抑制整合素的表达。实验分为对照组、诱导组和抑制组,BMSCs培养7 d和14 d后,分别采用免疫细胞化学(SABC法)检测II型胶原(col-II),甲苯胺蓝染色检测蛋白多糖的表达情况,Western Blot检测型胶原蛋白和相应整合素β_(1)蛋白含量的表达。结果:培养7 d和14 d后,诱导组col-II和蛋白多糖的表达均呈阳性,细胞形态由原来的长形或长梭形变短变圆,而对照组和抑制组均为阴性。诱导组col-II和相应整合素β_(1)蛋白的表达量均增多,而抑制组则较对照组减少,但两组的col-II表达均为阴性。结论:TGF-β3能上调整合素受体的表达,促进BMSCs的软骨分化;抗整合素β_(1)的单克隆抗体可以下调整合素β_(1)的表达,抑制BMSCs的软骨分化。Objective:To study and investigate the role of integrinβ1 in transforming growth factor(TGF)-induced differentia⁃tion of BMSCs into chondrocytes.Methods:Rat bone marrow stromal stem cells(BMSCs)were cultured in vitro using the whole bone marrow method.TGF-β3 promoted and induced chondrogenic differentiation of BMSCs and anti-integrinβ1 monoclonal anti⁃body inhibited the expression of integrins.The BMSCs were divided into control,induction and inhibition groups.7 and 14d after culture,type II collagen(col-II)was detected by immunocytochemistry(SABC method),proteoglycan expression was detected by to⁃luidine blue staining,and the expression of type II collagen and the corresponding integrinβ1 protein content was detected by Western Blot.Results:After 7 and 14 d of culture,the expression of col-II and proteoglycan in the induced group was positive,and the cell morphology changed from the original long shape or long shuttle shape to short and round,while both the control and inhibited groups were negative.The expression of col-II and the corresponding integrinβ1 protein was increased in the induction group,whereas it was decreased in the inhibition group compared with the control group,but col-II expression was negative in both groups.Conclusion:TGF-β3 upregulates integrin receptor expression and promotes chondrogenic differentiation of BMSCs.Monoclonal antibodies against integrinβ1 down-regulate integrinβ1 expression and inhibit cartilage differentiation in BMSCs.

关 键 词：骨髓基质干细胞 软骨分化 转化生长因子 整合素β_(1) 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]