摘要
目的:探讨镁离子转运蛋白1(MagT1)在脓毒症患者外周血CD8^(+)T淋巴细胞中的表达水平及其作用。方法:收集2018年3月至2021年5月于东莞市滨海湾中心医院就诊的96例脓毒症患者资料,根据感染及循环衰竭程度将脓毒症患者分为普通脓毒症组(n=39)、严重脓毒症组(n=33)和脓毒性休克组(n=24),另取同期健康者为对照组(n=30)。分离培养各组患者外周血CD8^(+)T细胞,检测外周血和CD8^(+)T细胞中Mg^(2+)浓度;qRT-PCR检测CD8^(+)T细胞中多种Mg^(2+)转运体(TRPM7、TRPM6、Mrs2p、SLC41A1、SLC41A2、MagT1及ACDP2)mRNA表达水平;Western blot检测CD8^(+)T细胞中MagT1表达水平;流式细胞术检测CD8^(+)T细胞表面抑制受体PD-1、Tim-3和活化受体NKG2D、HLA-DR表达水平;Pearson相关性分析MagT1与PD-1、Tim-3、NKG2D及HLA-DR表达水平相关性。结果:与对照组比较,脓毒症患者CD8^(+)T细胞中Mg^(2+)浓度显著降低(P<0.05),且病情越重,浓度越低,而外周血中Mg^(2+)浓度差异无统计学意义(P>0.05)。与对照组比较,脓毒症患者CD8^(+)T细胞中仅MagT1 mRNA表达水平显著降低(P<0.05),而其他6种Mg^(2+)转运体mRNA表达水平差异无统计学意义(P>0.05)。与对照组比较,脓毒症患者CD8^(+)T细胞中MagT1表达水平也显著降低(P<0.05),且其细胞表面抑制分子PD-1和Tim-3表达水平显著增加(P<0.05),而活化分子NKG2D和HLA-DR表达水平显著降低(P<0.05)。Pearson相关分析显示,MagT1与PD-1和Tim-3呈显著负相关,而与NKG2D和HLA-DR呈显著正相关(P<0.05)。结论:MagT1在脓毒症患者CD8^(+)T细胞中低表达,且通过降低胞内Mg^(2+)浓度,调控CD8^(+)T细胞表面分子表达,导致脓毒症患者CD8^(+)T细胞耗竭。
Objective: To explore the expression and role of magnesium transporter 1(MagT1) in peripheral blood CD8^(+)T lymphocytes of patients with sepsis. Methods: The data of 96 patients with sepsis treated in Dongguan Binhaiwan Central Hospital from March 2018 to May 2021 were collected. According to the degree of infection and circulatory failure, the patients were divided into common sepsis group(n=39), severe sepsis group(n=33) and septic shock group(n=24), and 30 healthy people from physical examination center were selected as control group during the same period. CD8^(+)T cells from the peripheral blood of patients in each group were isolated and cultured, and the Mg^(2+)concentration in peripheral blood and CD8^(+)T cells was detected. The expressions of variety of Mg^(2+)transporters(TRPM7, TRPM6, Mrs2 p, SLC41 A1, SLC41 A2, MagT1 And ACDP2) mRNA in CD8^(+)T cells were determined by qRT-PCR assay. The expression of MagT1 protein in CD8^(+)T cells was detected by Western blot method. The expression levels of CD8^(+)T cell surface inhibitory receptors PD-1, Tim-3 and activated receptors NKG2 D, HLA-DR were detected by flow cytometry. Pearson correlation method was used to analyze the correlation between the MagT1 and the PD-1, Tim-3, NKG2 D and HLA-DR expression levels. Results: Compared with the control group, the Mgconcentration in CD8^(+)T cells of sepsis patients was significantly reduced(P<0.05), and the more severe the disease, the lower the concentration, while the Mg^(2+)concentration in peripheral blood had no significant difference(P>0.05). The expressions of MagT1 mRNA in CD8^(+)T cells of sepsis patients were significantly reduced(P<0.05), but there was no significant difference in the mRNA expression of the other six Mg^(2+)transporters(P>0.05).The expression of MagT1 protein in CD8^(+)T cells of sepsis patients was also markedly decreased(P<0.05), and the expressions of cell surface inhibitory molecules PD-1 and Tim-3 were significantly increased(P<0.05), while the expressions of activation molecules NKG2 D and HLA-DR were significantly decreased(P<0.05). Pearson correlation analysis showed that MagT1 was significantly and negatively correlated with PD-1 and Tim-3, but was significantly and positively correlated with NKG2 D and HLA-DR(P<0.05). Conclusion: MagT1 is low expressed in CD8^(+)T cells in patients with sepsis, and it regulates the expression of CD8^(+)T cell surface molecules by reducing the intracellular Mg^(2+)concentration, which leads to the depletion of CD8^(+)T cells in patients with sepsis.
作者
黄文丽
黄冬枚
杨道贵
HUANG Wenli;HUANG Dongmei;YANG Daogui(Department of Laboratory Sciences,Dongguan Binhaisvan Central Hospital,Dongguan 523900,China)
出处
《现代医学》
2022年第5期549-554,共6页
Modern Medical Journal
基金
东莞市社会科技发展项目(201750715025152)。
