摘要
目的探讨T细胞免疫球蛋白黏蛋白分子-3(Tim-3)/半乳糖凝集素-9(galectin-9)在脂多糖(LPS)诱导淋巴细胞炎性过程中对辅助性T细胞(Th)Th1/Th2细胞失衡中的作用。方法收集脓毒症患者的外周血。分离外周血单个核细胞(PBMC),RT-qPCR检测Tim-3、galectin-9、T-bet和GATA3的表达;ELISA检测血清sTim-3、galectin-9、IFN-γ和IL-4的水平;分离的外周血单个核细胞体外培养,LPS刺激建立体外炎性模型;ELISA法检测细胞培养上清中Th1和Th2细胞相关细胞因子的水平;Western blot检测JAK2/STAT3相关因子的表达;Pearson相关系数分析Tim-3、galectin-9、IFN-γ和IL-4的相关性。结果脓毒症患者外周血中Tim-3和galectin-9的mRNA表达增多,sTim-3、galectin-9、IL-4和GATA-3升高,IFN-γ和T-bet水平下降,磷酸化JAK2和STAT3升高(P<0.05)。阻断Tim-3表达后,IFN-γ水平升高,而IL-4的水平明显下降,磷酸化JAK2和STAT3下降(P<0.05)。Tim-3、IL-4与galectin-9呈正相关,而Tim-3与IFN-γ呈负相关,IFN-γ与galectin-9呈负相关,Tim-3与IL-4呈正相关。结论在LPS诱导的炎性细胞中,Tim-3/galectin-9可能调控JAK2/STAT3通路,导致Th1/Th2细胞的失衡。
Objective To investigate the mechanism of Tim-3/galectin-9 in T helper type 1/2(Th1/Th2)cell imbalance in lipopolysaccharide(LPS)-induced human lymphocytes.Methods Peripheral blood mononuclear cells(PBMC)were isolated from patients with sepsis.The expression of Tim-3,galectin-9,T-bet and GATA3 were detected by RT-qPCR;ELISA was used for detecting sTim-3,galectin-9,IFN-γand IL-4.Isolated peripheral blood mononuclear cells were incubated with LPS and then cytokines related to Th1 and Th2 cells in the supernatant of cell culture were detected by ELISA;Western blot was used to detect the expression of JAK2/STAT3 related factors;Pearson correlation coefficient analysis was performed to identify relationship among Tim-3,galectin-9,IFN-γand IL-4.Results The mRNA expression of Tim-3 and galectin-9 in peripheral blood of patients with sepsis was increased,and protein expression of Tim-3,galectin-9,IL-4 and GATA-3 was increased significantly.The levels of IFN-γand T-bet were decreased and levels of p-JAK2 and p-STAT3 were elevated.After blocking Tim-3 expression,IFN-γlevel increased,while IL-4 level decreased significantly.Phosphorylation of JAK2 and STAT3 decreased.Level of Tim-3 and of IL-4 was positively correlated with galectin-9,while Tim-3 was negatively correlated with IFN-γ;IFN-γwas negatively correlated with galectin-9,and Tim-3 was positively correlated with IL-4.Conclusions Tim-3/galectin-9 regulates the JAK2/STAT3 pathway and leads to the imbalance of Th1/Th2 cells in LPS induced inflammation.
作者
孙晓丽
张进进
宋凤英
黄永莉
陈立力
邢颜超
SUN Xiaoli;ZHANG Jinjin;SONG Fengying;HUANG Yongli;CHEN Lili;XING Yanchao(Department of Blood Transfusion,General Hospital of Xinjiang Military Command,Urumqi 830000;Department of Clinical Laboratory,Yumin County People's Hospital,Yumin 834700,China)
出处
《基础医学与临床》
2023年第10期1542-1548,共7页
Basic and Clinical Medicine
基金
新疆维吾尔自治区自然科学基金青年科学基金(2022D01C843)。
