关键词: 非酒精性脂肪性肝病, 非酒精性脂肪肝, 非酒精性脂肪性肝炎, iNKT, α-GalCer

Abstract: To investigate the effect of intraperitoneal injection of α-GalCer on non-alcoholic fatty liver- DOI:10.3969/j.issn.1000-1565.2021.06.007腹腔注射α-GalCer对非酒精性脂肪性肝病的影响梁蕊^1,2,高翔^1,2,滕景芳^1,2,王建国³,程树杰^3,4,孟明^1,2(1.河北大学 基础医学院,河北 保定 071000; 2.河北省炎性自身免疫性疾病发病机制及防治重点实验室,河北 保定 071000; 3.河北大学附属医院 肝胆外科,河北 保定 071000; 4.河北省普通外科数字医学基础研究重点实验室,河北 保定 071000)摘 要:探讨了腹腔注射α-GalCer对非酒精性脂肪性肝病(NAFLD)的影响,为以iNKT细胞为靶点的NAFLD免疫学防治奠定基础.利用高脂饲料或蛋氨酸/胆碱缺乏饲料分别喂养C57BL/6J小鼠,建立非酒精性脂肪肝(NAFL)模型和非酒精性脂肪性肝炎(NASH)模型.全自动生化分析仪分析小鼠外周血中TCH、TG、HDL、LDL、ALT及AST水平;H&E染色观察肝脏病理变化;流式细胞术检测肝脏iNKT、iNKT1、iNKT2细胞频率.与对照组相比,NAFL小鼠体质量升高,血清TCH、HDL、LDL及ALT水平升高,肝脏脂肪沉积增加;而NASH小鼠体质量降低,血清TCH、TG及LDL水平降低,肝脏脂肪沉积增加并伴有炎细胞浸润.腹腔注射α-GalCer后,NAFL小鼠血清TCH、HDL及LDL水平降低,肝脏脂肪沉积减少,肝脏iNKT2亚群比例增加;NASH小鼠血清ALT和AST水平升高,肝脏脂肪沉积减少,炎细胞浸润增加,肝脏iNKT1、iNKT2亚群比例增加,其中以iNKT1亚群增加为主.腹腔注射α-GalCer可以改善NAFL和NASH小鼠肝脏脂肪沉积;小鼠NAFLD发病不同阶段(NAFL或NASH)肝脏局部微环境的改变可能影响α-GalCer对肝脏iNKT细胞不同亚群的激活,为进一步研究NAFLD的发病机制及免疫学治疗策略提供了有价值的实验数据.关键词:非酒精性脂肪性肝病;非酒精性脂肪肝;非酒精性脂肪性肝炎;iNKT;α-GalCer中图分类号:R392.5 文献标志码:A 文章编号:1000-1565(2021)06-0672-10Effect of intraperitoneal injection of α-GalCer on nonalcoholic fatty liver diseaseLIANG Rui^1,2, GAO Xiang^1,2, TENG Jingfang^1,2, WANG Jianguo³, CHENG Shujie^3,4, MENG Ming^1,2(1. College of Basic Medicine, Hebei University, Baoding 071000, China;2. Key Laboratory of Pathogenesis Mechanism and Control of Inflammatory-autoimmune Diseases in Hebei Province, Baoding 071000, China;3. Department of Hepatobiliary Surgery, Affiliated Hospital of Hebei University, Baoding 071000, China;4. Key Laboratory of General Surgery Digital Medicine Basic Researchin Hebei Province, Baoding 071000, China)Abstract: To investigate the effect of intraperitoneal injection of α-GalCer on non-alcoholic fatty liver- 收稿日期:2021-05-23 基金项目:国家自然科学基金面上项目(81771755);河北省自然科学基金资助项目(H2020201022;H2020201300) 第一作者:梁蕊(1995—),女,河北沧州人,河北大学在读硕士研究生,主要从事非酒精性脂肪性肝病研究.E-mail: liangrui1208@163.com 通信作者:孟明(1964—),女,辽宁义县人,河北大学教授,博士,博士生导师,主要从事自身免疫性疾病及免疫药理学研究.E-mail: mengming127@163.com程树杰(1965—),男,河北安国人,河北大学教授,博士生导师,主要从事肝胆胰脾疾病等研究.E-mail: shujiecheng@yeah.net第6期梁蕊等:腹腔注射α-GalCer对非酒精性脂肪性肝病的影响disease, and to lay a foundation for immunological prevention and treatment of non-alcoholic fatty liver disease(NAFLD)targeting iNKT cells. Nonalcoholic fatty liver disease(NAFL)model and nonalcoholic steatohepatitis(NASH)model were established by feeding C57BL/6J mice with high fat diet or methionine/choline deficiency diet. The levels of TCH, TG, HDL, LDL, ALT and AST in the peripheral blood of mice were analyzed by automatic biochemical analyzer. The pathological changes of liver were observed by H&E staining. The cell frequencies of iNKT, iNKT1 and iNKT2 in liver were detected by flow cytometry. Compared with the control group, the body weight of NAFL mice increased, the serum levels of TCH, HDL, LDL, ALT increased, and the hepatic fat deposition increased. While the body weight of NASH mice decreased, the serum levels of TCH、TG and LDL decreased, liver fat deposition is increased with inflammatory cell infiltration. After intraperitoneal injection of α-GalCer, serum levels of TCH, HDL, LDL decreased, liver fat deposition decreased,and the proportion of liver iNKT2 subsets increased in NAFL mice. While the serum levels of ALT and AST increased, liver fat deposition decreased, inflammatory cell infiltration increased and the proportion of liver iNKT1 and iNKT2 subsets increased, but mainly iNKT1 subsets in NASH mice. Intraperitoneal injection α-GalCer can improve NAFL and NASH mice liver fat deposition; changes in local microenvironment in different stages of NAFLD in mice(NAFL or NASH)may affect the activation of α-GalCer to different subgroups of liver iNKT cells, which provides valuable experimental data for further study of the pathogenesis and immunological treatment strategies of NAFLD.

Key words: NAFLD, NAFL, NASH, iNKT, α-GalCer