Glt25d1基因通过调节NKT细胞及Treg细胞比例加重Con A诱导的免疫性肝损伤  

Glt25d1 gene aggravates Con A-induced immune liver injury by regulating the ratio of NKT cells and Treg cells

作　　者：韦何锐 高美欣 肖凡 魏红山 Wei Herui;Gao Meixin;Xiao Fan;Wei Hongshan(Department of Gastroenterology,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Department of Gastroenterology,Beijing Tongren Hospital,Capital Medical University,Beijing 100176,China;Beijing Key Laboratory of Emerging Infectious Diseases,Institute of Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Beijing Institute of Infectious Diseases,Beijing 100015,China;National Center for Infectious Diseases,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,Beijing 100015,China)

机构地区：[1]首都医科大学附属北京地坛医院消化科,北京100015 [2]首都医科大学附属北京同仁医院消化科,北京100176 [3]首都医科大学附属北京地坛医院传染病研究所,新发突发传染病研究北京市重点实验室,北京100015 [4]北京市感染性疾病研究中心,北京100015 [5]国家传染病医学中心,首都医科大学附属北京地坛医院,北京100015 [6]传染病溯源预警与智能决策全国重点实验室,北京100015

出　　处：《中国肝脏病杂志（电子版）》2023年第4期33-38,共6页Chinese Journal of Liver Diseases：Electronic Version

基　　金：国家自然科学基金项目(82170541);北京市自然科学基金项目(7202071)。

摘　　要：目的 探讨胶原β(1-O)半乳糖基转移酶[collagen β(1-O) galactosyltransferase 1,COLGALT1/GLT25D1]基因在刀豆蛋白A(concanavalin A,Con A)诱导的自身免疫性肝损伤中的作用及可能机制。方法 6~8周雌性野生型(wild type,WT)小鼠和Glt25d1基因敲低杂合子(Glt25d1^(+/-))小鼠分别分为2组(对照组和Con A造模组),每组中WT小鼠和Glt25d1^(+/-)小鼠均各有6只。Con A以10 mg/kg体质量的剂量经内眦静脉丛给药。于造模后12 h处死小鼠。留取血浆,检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天门冬氨酸氨基转移酶(aspartate transaminase,AST)水平,留取肝组织,用于肝组织病理学评估和肝脏GLT25D1蛋白表达量测定。制备肝脏、脾脏及外周血单细胞悬液,运用流式细胞术比较两种小鼠的调节性T(regulatory T,Treg)细胞、自然杀伤T(natural killer T,NKT)细胞比例,并比较NKT细胞表面凋亡相关因子配体(factor related apoptosis ligand,FasL)表达差异。结果 对照组和Con A造模组中Glt25d1^(+/-)小鼠肝组织GLT25D1表达量均显著低于WT小鼠(对照组:0.342±0.168 vs1.144±0.169,t=5.841,P=0.004;Con A造模组:0.264±0.087 vs 0.964±0.058,t=11.640,P=0.0003)。经Con A诱导12 h后,Glt25d1^(+/-)小鼠ALT水平显著高于WT小鼠[(10155.2±4489.5)U/L vs (3078.5±111.0)U/L;t=-3.522,P=0.024]。肝组织HE染色显示Glt25d1^(+/-)小鼠肝板结构损坏更为严重,肝脏坏死区域更加广泛,汇管区炎性细胞浸润更加显著。流式细胞术检测显示,Con A造模组中Glt25d1^(+/-)小鼠脾脏中Treg细胞频数较WT小鼠显著减少[(7.849±1.116)%vs (9.892±1.762)%;t=2.978,P=0.008],肝脏NKT细胞频数[(9.244±6.898)%vs (3.376±4.794)%;t=-2.253,P=0.037]和NKT细胞FasL表达量[(29.62±4.960)%vs (16.43±3.964)%;t=-5.027,P=0.001]显著高于WT小鼠。结论 Glt25d1基因敲低加重Con A诱导的免疫性肝损伤,其机制可能与其降低Treg细胞频数、增加NKT细胞频数及增强NKT细胞功能有关。Objective To investigate the role of Collagenβ(1-O)galactosyltransferase 1(COLGALT1/GLT25D1)on concanavalin A(Con A)induced autoimmune liver injury and its potential mechanism.Methods 6~8 weeks female wild type(WT)mice and Glt25d1 gene knockdown heterozygous(Glt25d1^(+/-))mice were randomly divided into two groups(control group and Con A administration group),respectively,six WT and Glt25d1^(+/-) mice were included in each group,respectively.Con A was administered via the internal iliac venous plexus at a dose of 10 mg/kg body mass.Mice were sacrificed after Con A challenged for 12 h.Plasma were collected to detect the level of serum alanine aminotransferase(ALT)and aspartate transaminase(AST).Liver tissues were collected to assess the liver histopathology and detect the expression level of GLT25D1 protein.Liver,spleen and peripheral blood single-cell suspension were prepared to compare the percentage of regulatory T(Treg)cells and natural killer T(NKT)cells from different mice,as well as the level of factor related apoptosis ligand(FasL)expressed in NKT cell surface.Results In both groups,the expression level of GLT25D1 in liver of Glt251d1^(+/-)mice was significantly lower than that in WT mice,respectively(control group:0.342±0.168 vs 1.144±0.169,t=5.841,P=0.004;Con A induced group:0.264±0.087 vs 0.964±0.058,t=11.640,P=0.0003).After Con A challenged for 12 h,the serum ALT level of Glt251d1^(+/-)mice was significantly higher than that of WT mice[(10155.2±4489.5)U/L vs(3078.5±111.0)U/L;t=-3.522,P=0.024].The results of liver tissues after HE staining showed that compared with WT mice,Glt251d1^(+/-)mice exhibited more serious liver plate structure damage,more extensive the liver necrosis area and more significant infiltration of inflammatory cells in the portal area.The results of flow cytometry showed that compared with WT mice,the Treg cells in spleen of Glt25d1^(+/-)mice in Con A induced group decreased significantly[(7.849±1.116)%vs(9.892±1.762)%;t=2.978,P=0.008],NKT cells[(9.244±6.898)%vs(3.376±4.79

关 键 词：Glt25d1基因 自身免疫性肝炎 调节性T细胞 自然杀伤T细胞 

分 类 号：R575[医药卫生—消化系统]