高胆红素血症对新生仔鼠sPD-L1表达水平及T淋巴细胞免疫功能的影响  

Effects of hyperbilirubinemia on the expression level of sPD-L1 and immune function of T lymphocytes in neonatal rats

作　　者：林柏杏 杨英桂 朱华民 LIN Baixing;YANG Yinggui;ZHU Huamin(Department of Neonatology,Shenzhen Hospital of Southern Medical University,Shenzhen,Guangdong 518100,China;Department of Urology,Shenzhen Hospital of Southern Medical University,Shenzhen,Guangdong 518100,China;Department of Hematology,Shenzhen Hospital of Southern Medical University,Shenzhen,Guangdong 518100,China)

机构地区：[1]南方医科大学深圳医院新生儿科,广东深圳518100 [2]南方医科大学深圳医院泌尿外科,广东深圳518100 [3]南方医科大学深圳医院血液内科,广东深圳518100

出　　处：《重庆医学》2023年第11期1601-1605,1610,共6页Chongqing medicine

基　　金：国家自然科学基金项目(81902788);广东省自然科学基金项目(2020A1515010902);深圳市宝安区医疗卫生基础研究立项(2020JD029)。

摘　　要：目的探讨高胆红素对新生SD仔鼠血清可溶性程序性死亡因子及其配体(sPD-L1)表达水平和T淋巴细胞不同亚群的调节作用,证实胆红素诱导表达sPD-L1,促进淋巴细胞的凋亡。方法取3 d龄新生SD鼠24只,将其随机平均分为4组:对照组、实验1组、实验2组、实验3组,分别向其腹腔注射生理盐水及胆红素25、50、100μg/g。取新生SD仔鼠血,采用Ficoll密度梯度离心法,使用EasySep免疫磁珠分选系统分离外周血单个核细胞(PBMC)中的T淋巴细胞。流式细胞分析术(FAM)检测不同水平胆红素处理后的新生SD仔鼠PBMC中的T淋巴细胞数(CD3^(+)、CD8^(+)、Foxp3^(+)细胞)及sPD-L1水平,并检测免疫细胞凋亡的情况。结果随着胆红素水平的增加,新生SD仔鼠血PBMC中CD3^(+)、CD8^(+)细胞逐渐减少,Foxp3^(+)细胞逐渐增加;死亡细胞(PI+细胞)和凋亡细胞(FITC+细胞)逐渐增加;sPD-L1水平增加。结论胆红素诱导表达sPD-L1,可抑制T细胞的免疫应答,促进T细胞的凋亡。Objective To explore the regulatory effect of hyperbilirubin on the expression level of soluble programmed cell death-ligand 1(sPD-L1)and different subsets of T lymphocytes in neonatal spraguedawley(SD)rats,and to confirm that bilirubin can induce the expression of sPD-L1 and promote the apoptosis of lymphocytes.Methods A total of 24 three days old SD rats were selected and randomly divided into four groups on average:the control group,group one,experimental group two and experimental group three,and were injected with saline and bilirubin at 25,50 and 100 ug/g respectively.The peripheral blood of rats was collected by Ficoll density gradient centrifugation and the EasySep immunomagnetic bead sorting system was used to separate T lymphocytes in peripheral blood mononuclear cells(PBMC).Flow cytometry(FCM)was used to detect the number of T lymphocytes(CD3^(+)cells,CD8^(+)cells,Foxp3^(+)cells)and the level of sPD-L1 in PBMC of neonatal SD rats treated with different levels of bilirubin,and to detect the apoptosis of immune cells.Results With the increase of bilirubin concentration,CD3^(+)cells and CD8^(+)cells in blood PBMC of neonatal SD rats decreased gradually,while Foxp3^(+)cells increased gradually.The number of dead cells(PI+cells)and apoptotic cells(FITC+cells)increased gradually.The level of sPD-L1 increased.Conclusion Bilirubin induces the expression of sPD-L1,which can inhibit the immune response of T cells,and promotes the apoptosis of T cells.

关 键 词：高胆红素血症 可溶性程序性死亡因子及其配体 T淋巴细胞 免疫应答 细胞凋亡 

分 类 号：R722.1[医药卫生—儿科]