摘要
目的:探讨趋化因子Fractalkine(FKN)对狼疮性肾炎(LN)小鼠Treg细胞凋亡的影响及机制。方法:构建LN小鼠模型,使用FKN中和抗体(Anti-FKN)、p38MAPK信号通路的抑制剂(SB203580)、p38MAPK信号通路的激活剂(U-46619)作用于正常小鼠及模型小鼠,采用免疫组化检测小鼠肾组织中FKN、磷酸化p38(p-p38)及Foxp3的蛋白表达;采用免疫磁珠法分选模型小鼠脾脏CD4^(+)CD25^(+)Treg细胞,使用腺病毒转染细胞的方式敲低FKN的表达水平,同时使用p38MAPK信号通路抑制剂(SB203580)及激活剂(U-46619)干预细胞。采用Western blot验证FKN敲低的成功转染、检测p38、p-p38、Foxp3、凋亡相关因子(Bax、Bcl-2及Cyt-c)的蛋白表达。结果:LN小鼠肾组织中的FKN及磷酸化p38MAPK的表达明显上调,低表达FKN可降低LN小鼠肾组织及Treg细胞中FKN和p-p38的表达,增加Foxp3的表达,同时增加细胞内抗凋亡因子Bcl-2蛋白的表达,而减少促凋亡因子Bax及Cyt-c蛋白的表达。p38MAPK信号通路抑制剂对上述因子的作用与低表达FKN相似,其激活剂可以逆转这些因子在LN小鼠Treg细胞中的表达。结论:FKN可能通过激活p38MAPK信号通路参与调控LN小鼠Treg细胞凋亡,为阐明LN的发病机制提供了实验依据。
Objective:To investigate the effect of Fractalkine(FKN)on apoptosis of Treg cells in mice with lupus nephritis(LN).Methods:The LN mice model was established.The Anti-FKN,p38MAPK signal pathway inhibitors(SB203580)and activator of p38MAPK signaling pathway(U-46619)were used to act on the normal mice and the model mice.The protein expressions of FKN,phosphorylated p38(p-p38)and Foxp3 were detected by immunohistochemistry;CD4^(+)CD25^(+)Treg cells in spleen were sorted by immunomagnetic bead method,and the expression level of FKN was knocked down by adenovirus transfection at the same time,the p38MAPK signal pathway inhibitor(SB203580)and activator(U-46619)were used to intervene the cells.Western blot was used to verify the successful transfection of FKN knockdown,and the protein expression of p38,p-p38,Foxp3 and apoptosis related factors(Bax,Bcl-2 and Cyt-c)were detected by Western blot.Results:Downregulation of FKN decreased the expression of FKN and p-p38,increased the expression of Foxp3,increased the expression of anti-apoptotic factor Bcl-2,but decreased the expression of pro-apoptotic factors Bax and Cyt-c.The effect of p38MAPK signaling pathway inhibitor on the above-mentioned factors was similar to that of FKN,and its activator could reverse the expression of these factors in Treg cells of LN mice.Conclusion:FKN can regulate the apoptosis of Treg cells in LN mice,which may be involved in the activation of p38MAPK signaling pathway,and provides experimental basis for elucidating the pathogenesis of LN.
作者
马敬雪
巩奇明
潘秀虹
何林檩
尤燕舞
MA Jingxue;GONG Qiming;PAN Xiuhong;HE Linlin;YOU Yanwu(Department of Nephrology,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第24期2951-2957,共7页
Chinese Journal of Immunology
基金
国家自然科学基金项目(H100881860296)
广西自然科学基金项目(2017GXNSFDA198005)资助。
