高脂饮食对自身免疫性肝炎肝损伤与焦亡的研究  

The effects of high-fat diet on liver damage and pyroptosis in autoimmune hepatitis

作　　者：陈璐[1] 陆峰彬 叶韦玮[2] 陈永平[1] 陈达之 吴锦露 胡恩德 李慧[1] 黄瑜 许烂漫[1] 李骥[1] Chen Lu;Lu Fengbin;Ye Weiwei;Chen Yongping;Chen Dazhi;Wu Jinlu;Hu Ende;Li Hui;Huang Yu;Xu Lanman;Li Ji(Department of Infectious Diseases, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou Key Laboratory of Hepatology, Hepatology Institute of Wenzhou Medical University, Wenzhou 325025, Chin)

机构地区：[1]温州医科大学附属第一医院感染内科温州市肝脏病学重点实验室,325025 [2]义乌市中心医院感染科 [3]浙江大学医学院附属第一医院传染病诊治国家重点实验室

出　　处：《中华传染病杂志》2018年第1期18-24,共7页Chinese Journal of Infectious Diseases

基　　金：基金项目：国家自然科学基金（81570514,81500477）;浙江省自然科学基金（LY15H030017,LQ15H030006,LQ17H030005）;温州市科技局（Y20150014）;北京医学奖励基金（YJHYXKYJJ-162）;温州市终末期肝病的预警与干预治疗科技创新团队（C20150005）;“十二五”科技重大专项（2013ZX10005002-001-008,2013ZX10002003,2012ZX10002004）

摘　　要：目的探讨高脂饮食（high-fat diet, HFD）对小鼠自身免疫性肝炎（autoimmune hepatits, AIH）肝脏损伤的影响。方法50只C57BL/6雄性小鼠选取6只为S-100制备组，剩余小鼠用随机数字法随机分成标准饮食组（standard chow, SC）、HFD组、AIH＋SC组及AIH＋HFD组，喂饲1周后进行AIH造模，4周后处死小鼠，收集小鼠肝脏、脾脏、血清，进行HE染色观察肝组织病理学改变；检测小鼠血清中ALT和AST变化；蛋白质印迹法和实时PCR法检检测肝组织中Nod样受体蛋白3（NLR pyrin domain containing 3, NLRP3）和含半胱氨酸的天冬氨酸蛋白水解酶1（cysteinyl aspartate specific proteinase, Caspase-1）的表达；ELISA试剂盒检测血清中TNF-α、IL-6和IL-1β的分泌量；流式细胞仪分析脾脏中Th17细胞的数量。各组间均数比较采用单因素方差分析（one-way ANOVA），每两组间比较方差齐性采用SNK-q检验，方差不齐采用非参数检验。结果HFD组肝组织可见少许脂肪空泡形成，AIH＋SC组可见血管周围炎性反应，AIH＋HFD组可见炎性反应加重，出现少许空泡样脂肪变性。与SC组小鼠相比，AIH＋SC组和HFD组小鼠的血清ALT和AST水平有所增加，AIH＋HFD组上升最为明显，各组均数相比差异均具有统计学意义（F值分别为57.12和37.58，P均〈0.05）。SC组外周血中Th17占CD4＋T细胞的比例为（2.98±0.90）%，HFD组为（6.89±0.99）%，AIH＋SC组为（6.47±1.08）%，AIH＋HFD组为（9.96±0.83）%，各组之间比较及两两比较差异有统计学意义（F＝54.05，P〈0.05）。SC组各血清细胞因子浓度分别为IL-1β [（7.62±2.81） ng/L]、IL-6 [（106.54±53.08） ng/L]、TNF-α [（107.26±36.20） ng/L]；HFD组分别为IL-1β [（25.06±7.09） ng/L]、IL-6 [（220.11±47.41） ng/L]、TNF-α [（273.77±33.62） ng/L]；AIH＋SC组分别为IL-1β [（17.49±5.68） ng/L]、IL-6[（260.73±50.29） ng/L]、TNF-α [（250.49±81.63） ng/L]；AIH＋HFD分别为IL-ObjectiveTo investigate the effect of high-fat diet （HFD） on liver damage caused by autoimmune hepatitis （AIH） in mice.MethodsFifty C57BL/6 male mice were divided randomly into four groups： standard chow （SC） group, HFD group, AIH ＋ SC group and AIH＋ HFD group. AIH model was built after feeding for one week and all mice were sacrificed after four weeks. Liver and spleen tissues and serum were collected. Liver histopathology was detected by HE staining. Serum alanine aminotransperase （ALT） and aspartate aminotransferase （AST） levels were measured. Western blot analysis and real-time polymerase chain reaction （PCR） analysis were used to test the expressions of NLR pyrin domain containing 3 （NLRP3） and cysteinyl aspartate specific proteinase 1 （Caspase-1）. The concentrations of interleukin （IL）-6, IL-1β and tumor necrosis factor （TNF）-α were analyzed using enzyme linked immunosorbent assay technology. The amount of Th17 cells in spleen was analyzed by FACS. Means among groups were analyzed with one-way ANOVA. SNK-q analysis was used for groups with homogeneity of variance, while nonparametric test was used for groups with variance nonhomogeneity.ResultsHistologically, the H＆E staining of liver tissue from HFD group showed adipose degeneration, and there was inflammation around vessel in AIH＋ SC group. Moreover, in AIH＋ HFD group, the inflammation was more serious with mildly adipose degeneration. Compared with SC group, serum levels of ALT and AST increased in HFD group and AIH＋ SC group, and greatest increase was observed in AIH＋ HFD group. The differences were statistically significant （F=57.12 and 37.58, both P〈0.05）. The proportions of Th17 cells in SC group, HFD group, AIH＋ SC group and AIH＋ HFD group were （2.98±0.90）%, （6.89±0.99）%, （6.47±1.08）% and （9.96±0.83）%, respectively. The differences among all groups were statistically significant （F=54.05, P〈0.05）. The concentrations of IL-1β, IL-6 and TNF-α in each group

关 键 词：肝炎 自身免疫性 脂肪因子类 高脂饮食 炎症小体 细胞焦亡 

分 类 号：R575.1[医药卫生—消化系统]