胶原诱导性关节炎大鼠不同组织中免疫调控基因G蛋白亚基Alpha Q的表达特性及定位分析

Expression and localization of immunoregulatory gene G protein subunit Alpha Q in different tissues of collagen-induced arthritis rats

目的研究免疫调控基因G蛋白亚基Alpha Q(Gnαq)在胶原诱导性关节炎(CIA)鼠不同组织中的表达特性及定位。方法建立CIA大鼠模型,对CIA大鼠心、肝、脾、肺、肾和关节中Gnαq表达特性定位。通过实时荧光PCR和蛋白质印迹法分析不同试验组大鼠不同组织中Gnαq mRNA和蛋白水平的表达,免疫组织化学技术进行细胞定位。采用析因设计的方差分析进行统计学处理。结果成功构建CIA大鼠模型,Gnαq PCR产物序列同源性均为99.34%,Gnαq在CIA和健康大鼠的不同组织均有不同程度的表达。在CIA大鼠关节和肺中Gnαq mRNA(4.406±0.480和2.412±0.186)和蛋白水平(99.8±9.6和65.9±5.0)表达量显著高于其他组织中的表达量,差异有统计学意义(F=1244538.874,154377.357,P均<0.01)。Gnαq在CIA和对照组大鼠不同组织中均检测到阳性信号。在关节组织的FLS、内皮细胞、基质细胞和巨噬细胞中检测到强阳性信号。结论Gnαq参与RA炎性反应的发生和发展,推测Gnαq可能是治疗RA新的作用靶点。

Objective To investigate the expression and localization of the immunoregulatory gene G protein subunit Alpha Q(Gnαq)in different tissues of collagen-induced arthritis(CIA)rats.Methods We investigated the expression and localization of Gnαq in the heart,liver,spleen,lung,kidney,and joints of the CIA rat model.The Gnαq mRNA and protein expression levels in these tissues of rats from CIA and control groups were analyzed by real-time quantitative polymerase chain reaction(RT-qPCR)and western blotting,respectively.The cellular localization of Gnαq protein was determined by immunohistoche-mistry(IHC)assays.Factorial analysis of variance was used for statistical analysis.Results The CIA rat model was successfully built-up.The sequence homology of Gnαq PCR products was 99.34%.Gnαq was expressed at various levels in different tissues of rats from the groups.The levels of Gnαq mRNA(4.406±0.480 and 2.412±0.186)and protein(99.8±9.6 and 65.9±5.0)in the joints and lungs of CIA rats were significantly higher than those in other tissues,the difference was statistically significant(F=1244538.874,154377.357,P<0.01).Positive expression signals for Gnαq were detected in various tissues of CIA and control rats,among which strong positive signals were detected in joint fibroblast-like synoviocytes(FLSs),endothelial cells,stromal cells,and macrophages.Conclusion Our results further confirms the involvement of Gnαq in the onset and devel-opment of inflammatory responses in RA,suggesting that Gnαq may be a new therapeutic target for RA.