摘要
目的观察英夫利西单克隆抗体对自身免疫性肝炎(AIH)的预防效果并探讨其作用机制。方法经鼠尾静脉注射刀豆蛋白A(Con-A)建立小鼠AIH模型。将40只小鼠随机分为预防组和对照组,每组20只。预防组Con-A注射前1 h经鼠尾静脉注射途径给予20 mg/kg英夫利西单克隆抗体,对照组注射200μL PBS。分别于Con-A或PBS注射后3、8、12和24 h取血清,通过比色法检测血清AST、ALT水平。通过ELISA法检测血清细胞因子IL-6、IL-1β、IFN-γ、IL-4、IL-17A、IL-10和趋化因子配体10(CXCL10)水平。于Con-A或PBS注射后12 h取肝组织标本进行H-E染色,通过流式细胞术检测浸润至肝脏的淋巴细胞,通过实时荧光定量PCR检测T盒子转录因子(TBX21)、GATA结合蛋白3(GATA3)、RAR相关孤儿受体C(RORC)和CXCL10基因mRNA的表达水平。通过蛋白质印迹法检测肝脏CXCL10表达水平。统计学方法采用单因素方差分析和配对t检验。结果预防组Con-A注射后8、12和24 h的血清ALT、AST、IL-1β、IFN-γ水平均低于对照组[(545.8±190.3)U/L比(865.8±237.7)U/L、(947.6±267.9)U/L比(1448.0±403.5)U/L和(508.6±131.1)U/L比(976.6±207.6)U/L;(620.7±132.0)U/L比(952.9±106.8)U/L、(801.6±212.0)U/L比(1424.8±236.0)U/L和(632.1±117.8)U/L比(1008.3±187.5)U/L;(31.38±10.12)ng/L比(48.12±11.53)ng/L、(39.34±11.40)ng/L比(60.00±14.17)ng/L和(29.49±8.22)ng/L比(46.89±5.50)ng/L;(432.93±66.82)ng/L比(674.66±97.88)ng/L、(655.09±169.17)ng/L比(937.90±166.36)ng/L和(263.40±54.97)ng/L比(410.74±86.64)ng/L],差异均有统计学意义(t=2.350、2.308、4.263,4.374、4.860、3.806,2.440、2.541、3.939,4.560、2.660、3.210;P均<0.05)。预防组Con-A注射后3、8、12和24 h的血清IL-6水平均低于对照组[(1075.79±303.77)ng/L比(1914.48±317.80)ng/L、(1945.97±271.85)ng/L比(2100.80±378.42)ng/L、(1578.60±504.54)ng/L比(2525.40±406.55)ng/L和(1020.64±280.03)ng/L比(1582.00±311.96)ng/L],差异均有统计学意义(t=4.266、2.903、3.267、2.994,P均<0.05
Objective To observe the preventive effects of infliximab in autoimmune hepatitis(AIH)and to explore its mechanism.Methods The mice AIH model was established by injecting concanavalin A(Con-A)into the caudal vein.Forty mice were divided into prevention group and control group,with 20 mice in each group.The mice of prevention group were injected intravenously with infliximab(20 mg/kg)one hour before Con-A injection and the mice of control group were administrated with 200μL phosphate buffered saline(PBS).Serum was collected 3,8,12 and 24 h after Con-A/PBS injection.The serum level of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)was detected by colorimetry.The level of cytokine interleukin(IL)-6,IL-1β,interferon gamma(IFN-γ),IL-4,IL-17A,IL-10 and chemokine C-X-C motif ligand 10(CXCL10)was measured by enzyme-linked immunosorbent assay(ELISA).Liver samples were taken 12 h after Con-A/PBS injection for hematoxylin-eosin staining.Liver infiltrated lymphocytes were assessed by flow cytometry.The expression of T-box transcription factor 21(TBX21),GATA binding protein 3(GATA3),RAR related orphan receptor C(RORC)and CXCL10 at mRNA level was evaluated by quantitative real-time polymerase chain reaction(qRT-PCR).The expression of CXCL10 in liver was detected by Western blotting.Paired t test and one-way analysis of variance were used for statistic analysis.Results At 8,12,and 24 h after Con-A injection,the serum ALT level,AST level,IL-1βand IFN-γof prevention group were all lower than those of control group((545.8±190.3)U/L vs.(865.8±237.7)U/L,(947.6±267.9)U/L vs.(1448.0±403.5)U/L,(508.6±131.1)U/L vs.(976.6±207.6)U/L;(620.7±132.0)U/L vs.(952.9±106.8)U/L,(801.6±212.0)U/L vs.(1424.8±236.0)U/L,(632.1±117.8)U/L vs.(1008.3±187.5)U/L;(31.38±10.12)ng/L vs.(48.12±11.53)ng/L,(39.34±11.40)ng/L vs.(60.00±14.17)ng/L,(29.49±8.22)ng/L vs.(46.89±5.50)ng/L;and(432.93±66.82)ng/L vs.(674.66±97.88)ng/L,(655.09±169.17)ng/L vs.(937.90±166.36)ng/L,(263.40±54.97)ng/L vs.(410.74±86.64)ng/L),and the di
作者
孙孟宇
万昌山
吕梦楠
宋岩
马润芝
刘文天
Sun Mengyu;Wan Changshan;Lyu Mengnan;Song Yan;Ma Runzhi;Liu Wentian(Department of Gastroenterology and Hepatology,Tianjin Medical University General Hospital,Tianjin 300052,China;Hematopoietic Stem Cell Transplantation Center,Institute of Hematology and Blood Diseases Hospital,Chinese Academy of Medical Sciences,Tianjin 300020,China)
出处
《中华消化杂志》
CAS
CSCD
北大核心
2020年第3期192-198,共7页
Chinese Journal of Digestion
基金
天津市科学技术委员会慢性病防治科技重大专项(17ZXMFSY00210)。
