肿瘤坏死因子α对强直性脊柱炎患者间充质干细胞成骨分化的影响  

Effects of tumor necrosis factor alpha on osteogenic differentiation of mesenchymal stem cells derived from ankylosing spondylitis patient

作　　者：米汝佳 蔡兆鹏[3] 苏鸿君[2] 汤苏安 沈慧勇[1,3] MI Rujia;CAI Zhaopeng;SU Hongjun;TANG Su'an;SHEN Huiyong(Department of Orthopedics,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510120,China;Department of Orthopedics,No.8th Hospital,Sun Yat-sen University,Shenzhen 518000,China;Center for Biotherapy,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510120,China;Department of Orthopedics,Nanfang Hospital,Southern Medical University,Guangzhou 510120,China)

机构地区：[1]中山大学孙逸仙纪念医院骨外科,广州510120 [2]中山大学孙逸仙纪念医院生物治疗中心,广州510120 [3]中山大学附属第八医院骨外科,深圳518000 [4]南方医科大学南方医院骨外科,广州510120

出　　处：《免疫学杂志》2020年第3期254-259,共6页Immunological Journal

摘　　要：目的研究TNF-α对HDMSCs和ASMSCs成骨分化的影响及其区别。方法实验以HDMSCs作为对照,分别以无TNF-α成骨诱导液和含有1 ng/ml、2 ng/ml和20 ng/ml不同质量浓度TNF-α的成骨诱导液诱导HDMSCs和ASMSCs成骨分化,采用碱性磷酸酶和茜素红检测各组成骨分化情况;实时荧光定量PCR方法检测各组成骨标志分子OPG、Runx2、Osterix基因表达水平;Western blot检测成骨分化相关Smad通路和Wnt通路的激活水平。结果与无TNF-α处理相比,低质量浓度TNF-α(1 ng/ml和2 ng/ml)处理下ASMSCs成骨分化减弱(P<0.05),OPG、Runx2、Osterix mRNA水平降低(P<0.05),Smad通路及Wnt通路被抑制(P<0.05);而HDMSCs成骨分化无明显改变(P>0.05),上述分子及通路改变无显著差异(P>0.05);与无TNF-α处理相比,高质量浓度TNF-α(20 ng/ml)处理下HDMSCs和ASMSCs成骨分化均减弱(P<0.05)。结论低浓度TNF-α可抑制AS患者体内MSCs成骨分化;高浓度TNF-α则对正常人MSCs和AS患者MSCs都有抑制作用。Tumor necrosis factor alpha(TNF-α) is an important inflammatory factor in patients with ankylosing spondylitis(AS), but its influence on the osteogenic differentiation of mesenchymal stem cells from healthy donors(HDMSCs) and mesenchymal stem cells from AS(ASMSCs) is still unknown. This study was preformed to investigate the effects of TNF-α on osteogenic differentiation of HDMSCs and ASMSCs and their difference. HDMSCs were taken as control in the experiments, then TNF-α-free and TNF-α(1 ng/ml, 2 ng/ml and 20 ng/ml) medium were used to induce osteogenic differentiation of HDMSCs and ASMSCs;alkaline phosphatase and alizarin red were used to detect the composition of bone differentiation;RT-PCR was applied to detect the expression of bone markers OPG, Runx2 and Osterix mRNA levels;and Western blot was used to detect the activation levels of bone differentiation-related Smad pathway and Wnt pathway. Compared with TNF-α-free treatment, low concentration of TNF-α(1 ng/ml and 2 ng/ml) could decrease the osteogenic differentiation of ASMSCs(P<0.05), down-regulate the mRNA levels of OPG, Runx2, and Osterix significantly(P<0.05), inhibit Smad and Wnt pathways(P<0.05). However, low concentration of TNF-α induce no significant difference in the osteogenic differentiation of HDMSCs(P>0.05).Compared with TNF-α-free treatment, the osteogenic differentiation of HDMSCs and HDMSCs were reduced in treatment of high concentration TNF-α(20 ng/ml)(P<0.05). All these results indicate that low concentration of TNF-α can only inhibit the osteogenic differentiation of MSCs in patients with ankylosing spondylitis, while high concentration of TNF-α has inhibitory effects on HDMSCs and ASMSCs.

关 键 词：肿瘤坏死因子Α 强直性脊柱炎 间充质干细胞 成骨分化 

分 类 号：R392.1l[医药卫生—免疫学]