IL-18对人骨髓间充质干细胞成骨分化的作用  

Effect of IL-18 on differentiation of human bone marrow mesenchymal stem cells into osteoblasts

作　　者：倪飞飞 徐庆 王亚辉[3] 李建军[1] NI Feifei;XU Qing;WANG Yahui;LI Jianjun(Shengjing Hospital of China Medical University,Shenyang 110004,China;不详)

机构地区：[1]中国医科大学附属盛京医院,沈阳110004 [2]天津市天津医院 [3]安徽医科大学附属六安市人民医院

出　　处：《山东医药》2021年第13期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金面上项目(81971829)。

摘　　要：目的探讨IL-18对人骨髓间充质干细胞(hBMSC)的成骨分化作用。方法取生长良好的第3代hBMSC每孔2×10^(4)个接种于6孔培养板,并随机分成两组。于细胞汇合率为80%~90%时,对照组采用经典成骨诱导液培养,观察组采用经典成骨诱导液+质量浓度为100 ng/mL的重组人IL-18培养液培养。成骨分化诱导1、3、7 d后检测骨特异性转录因子-2(Runx2)、骨形态发生蛋白-2(BMP2)、骨桥蛋白(OPN)mRNA及蛋白表达。两组hBMSC诱导分化7 d后进行碱性磷酸酶(ALP)染色、茜素红染色。结果两组成骨诱导1、3、7 d,Runx2、BMP2、OPN mRNA及蛋白表达比较差异有统计学意义(P均<0.05)。观察组随诱导时间延长,Runx2、BMP2、OPN mRNA及蛋白表达增加,不同时点Runx2、BMP2、OPN mRNA及蛋白表达差异有统计学意义(P均<0.05)。观察组成骨诱导7d后ALP染色较对照组明显深染,茜素红染色矿化结节染色区域明显多于对照组。结论IL-18体外能促进hBMSC向成骨细胞分化。Objective To explore the effect of IL-18 on osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSC).Methods The well-growing third-generation hBMSCs were inoculated into 6-well culture plate with 2×10^(4)per well and then were randomly divided into two groups.When the cell confluence rate was 80%-90%,the cells in the control group were cultured with classical osteogenic induction liquid,and the cells in the observation group with classical osteogenic induction liquid+recombinant human IL-18 culture medium with a mass concentration of 100 ng/mL.The bone-specific transcription factor-2(Runx2),bone morphogenetic protein-2(BMP2),and osteopontin(OPN)mRNA and protein expression levels were detected at 1,3,and 7 days after osteogenic differentiation induction.The hBMSCs of the two groups were stained with alkaline phosphatase and alizarin red for 7 days after induction of differentiation.Results There were statistically significant differences in the mRNA and protein expression levels of Runx2,BMP2,and OPN between the two groups at 1,3,and 7 days after induction of differentiation(all P<0.05).In the observation group,over the time of induction,the expression levels of Runx2,BMP2,and OPN mRNA and protein increased,and the differences in Runx2,BMP2,and OPN mRNA and protein expression levels at different time points were statistically significant(all P<0.05).It was observed that at 7 days after osteoinduction,ALP staining was significantly darker in the observation than in the control group,and the staining area of mineralized nodules in alizarin red staining was significantly larger than that of the control group.Conclusion IL-18 can promote the differentiation of hBMSC into osteoblasts in vitro.

关 键 词：骨髓间充质干细胞 IL-18 骨特异性转录因子-2 骨形态发生蛋白-2 骨桥蛋白 成骨作用 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]