炎性环境下FSTL1对小鼠骨髓间充质干细胞成骨性能的影响  

Effect of FSTL1 on osteogenesis of mouse bone marrow mesenchymal cells in inflammatory environment

作　　者：金齐尧 毛广艳 邓威 王晨星[1] 李怀奇[1] 王东苗[1] 叶金海[1] JIN Qiyao;MAO Guangyan;DENG Wei;WANG Chenxing;LI Huaiqi;WANG Dongmiao;YE Jinhai(Jiangsu Key Laboratory of Oral Diseases, Department of Oral and Maxillofacial Surgery, Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing 210029, China)

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,南京医科大学附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《口腔生物医学》2020年第2期120-124,共5页Oral Biomedicine

基　　金：国家自然科学基金(81371123);“科教强卫工程”医学重点人才项目(ZDRCA2016087);江苏高校优势学科建设工程资助项目(2018-87)。

摘　　要：目的:研究炎性环境下卵泡抑素样蛋白1(FSTL1)对小鼠骨髓间充质干细胞(mBMSCs)成骨性能的影响。方法:将mBMSCs转染慢病毒,改变细胞FSTL1的表达,通过CCK-8法检测细胞增殖活性。转染慢病毒的mBMSCs经过10 ng/mL的肿瘤坏死因子α(TNF-α)刺激后,再予以成骨诱导;通过碱性磷酸酶(ALP)和茜素红染色实验,观察mBMSCs的成骨能力的变化;通过实时定量PCR检测细胞Ⅰ型胶原蛋白(Col1)、骨桥蛋白(Opn)、骨钙素(Ocn)等成骨相关基因的表达。结果:通过慢病毒转染能够有效地改变mBMSCs内Fstl1基因的表达,并且不影响细胞增殖活性(P>0.05)。在含有10 ng/mL TNF-α的炎症环境中,与对照组相比,过表达FSTL1的mBMSCs成骨诱导后ALP和茜素红染色变浅,成骨相关基因表达降低(P<0.05);而敲减FSTL1的mBMSCs成骨诱导后ALP和茜素红染色变深,成骨相关基因表达升高(P<0.05)。结论:FSTL1促进了炎症对mBMSCs成骨性能的抑制作用,抑制FSTL1能有效地减轻炎症对BMSCs的影响。Objective:To investigate whether follistatin-like protein 1(FSTL1)can affect the osteogenic performance of mouse bone marrow mesenchymal stem cells(mBMSCs)in an inflammatory environment.Methods:mBMSCs were transfected with lentivirus to change the expression of FSTL1,and the proliferation activity of these cells was detected by CCK-8 experiment.The mBMSCs transfected with lentivirus were stimulated with 10 ng/mL TNF-α,and then induced into osteoblasts.Alkaline phosphatase(ALP)and alizarin red staining experiments were used to detect the changes of osteogenic ability of mBMSCs;the expression of collagen typeⅠcollagen(Col1),osteopontin(Opn),osteocalcin(Ocn)was detected by real-time quantitative PCR.Results:Lentivirus transfection could effectively change the expression of FSTL1 gene in mBMSCs,and there was no significant difference in the proliferation activity of these cells(P>0.05).In the inflammatory environment containing 10 ng/mL TNF-α,compared with the control group,mBMSCs overexpressing FSTL1 had lighter ALP and alizarin red staining results after osteogenic induction,and the expression of Col1,Opn,and Opn genes decreased(P<0.05);mBMSCs knocked down FSTL1 had darker ALP and alizarin red staining results after osteogenic induction,and the expression of Col1,Opn,and Opn genes increased(P<0.05).Conclusions:FSTL1 promotes the inhibitory effect of inflammation on the osteogenic performance of mBMSCs,and inhibiting FSTL1 can effectively reduce the effect of inflammation on mBMSCs.

关 键 词：卵泡抑素样蛋白1 骨髓间充质干细胞 成骨分化 炎症 

分 类 号：R780.2[医药卫生—口腔医学]