BaP通过AhR抑制BMP2诱导的间充质干细胞C3H10T1/2成骨分化  

BaP Inhibits BMP2-Induced Osteogenic Differentiation of Mesenchymal Stem Cells Line C3H10T1/2 through AhR

作　　者：安利钦 施琼[1] 周一青[1] 刘红霞[1] 程瑜 张汝益[1] 严树涓[1] 翁亚光[1] An Liqin;Shi Qiong;Zhou Yiqing;Liu Hongxia;Cheng Yu;Zhang Ruyi;Yan Shujuan;Weng Yaguang(Key Laboratory of Clinical Laboratory Diagnostics of Ministry Education, Faculty of Laboratory Medicine, Chongqing Medicine University, Chongqing 400016, China)

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2018年第3期326-333,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81672103;31200971);重庆市渝中区科委科技项目(批准号:20150109)资助的课题~~

摘　　要：该文主要研究苯并芘(benzoapyrene,Ba P)对骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)介导的间充质干细胞C3H10T1/2细胞成骨分化的影响,并探究这种作用的调控机制。用腺病毒Ad-BMP2感染C3H10T1/2细胞,RT-PCR检测到BMP2的表达水平显著增高(P<0.001),芳香烃受体(aryl hydrocarbon receptor,Ah R)的表达则无显著差异。随后加入不同浓度Ba P处理,检测Ba P对BMP2诱导的C3H10T1/2细胞成骨分化的影响,处理7天检测碱性磷酸酶(alkaline phosphatase,ALP)的染色和活性,14天检测茜素红S染色。结果显示,Ba P可以剂量依赖的方式抑制BMP2介导的ALP和钙盐沉积(P<0.001);Western blot检测结果显示,Ba P可以剂量依赖的方式抑制BMP2诱导的p-Smad1/5/8(p-drosophila mothers against de-capentaplegic 1/5/8)及Runx2的表达(P<0.01,P<0.001)。应用Ah R拮抗剂CH223191后,测定结果显示,其可部分逆转Ba P对BMP2诱导的C3H10T1/2细胞早晚期成骨分化及BMP2/Smad信号通路的抑制作用(P<0.05,P<0.001)。该研究结果提示,Ba P可通过Ah R抑制BMP2介导的促进间充质干细胞C3H10T1/2细胞成骨分化,这一过程与BMP2/Smad信号通路受到抑制有关。The aim of this study was to investigate the effect of Benzoapyrene on bone morphogenetic protein 2 （BMP2）-induced osteogenic differentiation of mesenchymal stem cells line C3H 10T 1/2 and the regulatory mechanism involved. C3H10T1/2 cells were infected with Ad-BMP2/Ad-GFP and the expression of BMP2 and aryl hydrocarbon receptor （AhR） were detected by RT-PCR. The mRNA level of BMP2 significantly increased （P〈0.001）, however, the the mRNA level of AhR did not alter significantly. Different concentration of BaP was to treat C3H10T1/2 for 7 days, detected with alkaline phosphatase （ALP） activity and ALP staining, and for 14 days, detected with Alizarin red S staining to observe the effect of BaP on BMP2-induced osteogenic differentiation. The results showed that BaP inhibited BMP2-induced ALP activity （P〈0.001） and calcium deposition in a dosedependent manner. It was also found that BaP significantly reduced the protein levels of p-Smadl/5/8 and Runx2 （P〈0.01, P〈0.001）. However, when added AhR antagonists （CH223191）, we found that CH223191 could partly reverse the toxicologic effects of BaP on oesteogenic differentiation, at the mean while, it partly rescued the inhibition effects of BaP on BMP2/Smad signal pathway （P〈0.05）. We concluded that BaP can inhibit BMP2- induced osteogenic differentiation of C3H10T1/2 cells via AhR and it involved in BMP2/Smad signaling pathway.

关 键 词：苯并芘 骨形态发生蛋白2 C3H10T1/2细胞 芳香烃受体 成骨分化 

分 类 号：R68[医药卫生—骨科学]