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.
作者
金齐尧
毛广艳
邓威
王晨星
李怀奇
王东苗
叶金海
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)
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)。
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
follistatin-like protein 1
bone marrow mesenchymal stem cells
osteogenic differentiation
inflammation
