Cripto-1表达在异体输血诱导的巨噬细胞分化后免疫抑制的机制  

Mechanism of immune suppression following macrophage differentiation induced by allogeneic blood transfusion through Cripto-1 expression

作　　者：杨苗苗 屈林林 沈鑫 陆熠 杨柳 吴理淑 YANG Miaomiao;QU Linlin;SHEN Xin;LU Yi;YANG Liu;WU Lishu(Department of Medical Laboratory,The First People's Hospital of Suqian,Suqian 223800,Jiangsu,China;Department of Pediatrics,The First People's Hospital of Suqian,Suqian 223800,Jiangsu,China;Department of Blood Transfusion,Fuding Hospital,Fuding 355200,Fujian,China)

机构地区：[1]宿迁市第一人民医院医学检验科,江苏宿迁223800 [2]宿迁市第一人民医院儿科,江苏宿迁223800 [3]福鼎市医院输血科,福建福鼎355200

出　　处：《西部医学》2025年第2期184-190,共7页Medical Journal of West China

基　　金：江苏省卫生健康委员会科研项目(20210368)。

摘　　要：目的 探讨Cripto-1对异体输血诱导巨噬细胞分化后免疫抑制的改善作用。方法 选取BALB/c和C57BL/6小鼠,进行异体输血,培养其巨噬细胞;建立稳定过表达Cripto-1的L02细胞系,与巨噬细胞共培养;Elisa和RT-PCR检测抗炎细胞因子IL-10和促炎细胞因子[肿瘤坏死因子α(TNF-α)、IL-6、转化生长因子β(TGF-β)和IL-1β]的蛋白表达和mRNA水平;蛋白印迹法检测NF-kB通路蛋白表达情况。结果 蛋白印迹检测在实验细胞培养上清液的细胞质中检测到Cripto-1。在L02细胞培养上清液的细胞质中检测到Cripto-1。暴露于Cripto-1上清液8 h或12 h可增加IL-1β和TNF-α的mRNA表达水平(P<0.05),但TGF-β的mRNA表达水平不升高(P>0.05)。细胞系分泌到上清液中的Cripto-1导致IL-6和IL-10 mRNA表达水平增加(P<0.05)。暴露于这种上清液显著增加了IL-10和TNF-α,IL-6和IL-1β的mRNA和蛋白表达水平(P<0.05),但不影响TGF-β的表达(P>0.05)。Cripto-1处理细胞的核p65水平、ibb激酶(IKB)磷酸化和ibb磷酸化都更高(P<0.05)。抑制NF-kB信号可减少Cripto-1介导的巨噬细胞。结论 Cripto-1通过NF-kB信号通路增强异体输血诱导巨噬细胞吞噬活性,并上调抗和促炎细胞因子的产生。Objective To explore the amelioration effect of Cripto-1 on the immunosuppression of macrophage differentiation induced by allogeneic transfusion.Methods BALB/c and C57BL/6 mice were selected for allogeneic blood transfusion and their macrophages were cultured.A stable L02 cell line with Cripto-1 overexpression was established and co-cultured with macrophages.The protein expression and mRNA levels of anti-inflammatory cytokines IL-10 and pro-inflammatory cytokines(TNF necrosis factor α(TNF-α),IL-6,transforming growth factor β(TGF-β) and IL-1β) were detected by Elisa and RT-PCR.The expression of NF-kB pathway protein was detected by western blot.Results Blot detection of Cripto-1 was detected in the cytoplasm of the supernatant of experimental cell culture.Cripto-1 was detected in the cytoplasm of L02 cell culture supernatant.Supernatant on exposure to Cripto-18 h or 12 h could increase IL-1 beta and TNF-alpha mRNA expression level(P<0.05),but the TGF-beta mRNA expression level was not higher(P>0.05).The mRNA expression levels of IL-6 and IL-10 were increased by Cripto-1,which was secreted into the supernutriment of cell lines(P<0.05).Exposed to such supernatant significantly increased the supernatant fluid IL 10 and TNF-alpha,IL-6 and IL-1β mRNA and protein expression level(P<0.05),but did not influence the TGF-β expression(P>0.05).Cripto-1 treated cells with higher nuclear p65 levels,ibb kinase(IKB) phosphorylation,and ibb phosphorylation(P<005).Conclusion Cripto-1 enhance phagocytic activity of macrophages induced by allogeneic transfusion through NF-kB signaling pathway,and up-regulate the production of anti-inflammatory and pro-inflammatory cytokines.

关 键 词：Cripto-1 异体输血 巨噬细胞 免疫抑制 

分 类 号：R457.1[医药卫生—治疗学]