曲古抑菌素A对s-100诱导的小鼠自身免疫性肝炎的影响  

Effect of trichostatin A on s-lOO-induced autoimmune hepatitis in mice

作　　者：胡恩德 昊锦露 陈永平[1] 陈达之 李骥[1] 许烂漫[1] 陆峰彬 陈璐[1] 黄瑜 

机构地区：[1]温州医科大学附属第一医院感染内科,325025 [2]浙江大学传染病诊治国家重点实验室、传染病诊疗创新协作中心

出　　处：《中华传染病杂志》2017年第12期734-738,共5页Chinese Journal of Infectious Diseases

基　　金：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）;王宝恩肝纤维化研究基金（xjs001）;中华医学会科研项目（1503023061i）;温州市终末期肝病的预警与干预治疗科技创新团队（C20150005）

摘　　要：目的探讨组蛋白去乙酰化酶抑制剂——曲古抑菌素A（trichostatin A，TSA）对s-100诱导的小鼠自身免疫性肝炎的影响。 方法将26只6周龄C57BL/6小鼠随机分为对照组、模型组和TSA组，每组6只，另外8只小鼠用于提取肝组织s-100蛋白。模型组和TSA组小鼠腹腔注射s-100和完全弗氏佐剂，建立小鼠自身免疫性肝炎模型，第21天，TSA组小鼠连续7 d腹腔注射TSA 2 mg/（kg·d），对照组和模型组小鼠腹腔注射等量含1%二甲基亚砜的0.9%氯化钠溶液。检测各组小鼠血清ALT和AST水平，观察肝组织病理学。蛋白质免疫法检测肝组织内核因子-κB和乙酰化组蛋白H3；实时定量PCR检测肝组织内核因子-κB、HDAC3、TLR4及TNF-α mRNA水平；ELISA法检测血清TNF-α。各组间比较采用t检验。 结果对照组、模型组和TSA组小鼠血清ALT分别为（122.00±45.29）、（459.33±167.58）和（217.33±49.25） U/L，对照组、TSA组与模型组比较，差异均有统计学意义（t值分别为4.76、3.41，均P〈0.05）；AST分别为（127.83±18.55）、（389.67±87.14）和（249.50±71.72） U/L，与模型组比较，差异均有统计学意义（t值分别为7.20、3.04，均P〈0.05）。TSA减轻了s-100诱导的肝组织病理炎症；模型组小鼠肝组织中核因子-κB蛋白、核因子-κB mRNA、TNF-α mRNA、HDAC3 mRNA和TLR4 mRNA的相对表达量分别为2.43±0.42、9.51±0.36、10.53±0.74、2.90±0.22和4.50±0.73，较对照组1.28±0.49、1.28±0.49、1.06±0.14、1.72±0.73和1.01±0.31明显升高（t值分别为4.68、37.14、30.69、4.33和10.85，均P〈0.05），TSA组核因子-κB蛋白、核因子-κB mRNA、TNF-α mRNA、HDAC3 mRNA及TLR4 mRNA相对表达量分别为1.30±0.36、1.30±0.36、2.38±0.36、2.13±0.32和2.40±0.51，也明显低于模型组（t值分别为4.58、30.62、24.12、2.81、5.81，均P〈0.05）。对照组、模型组和TSA组小鼠血清TNF-α分别为（122.37±68.12）、（1 361.44±207.13）和�ObjectiveTo investigate the effect of trichostatin A （TSA）, a histone deacetylase inhibitor, on s-100-induced autoimmune hepatitis in mice. MethodsA total of 26 six-week-old male C57BL/6 mice were randomly divided into control group, model group and TSA group （six in each group）, and the rest 8 mice were used to extract the s-100 protein from liver tissue. Mice of model group and TSA group were injected intraperitoneally with s-100 with complete Freund＇s adjuvant to induce autoimmune hepatitis model. At day 21, TSA group mice were injected intraperitoneally with TSA 2 mg/（kg·d） for 7 days, and 0.9% sodium chloride solution containing 1% dimethyl sulfoxide was injected into the control and model group mice. Alanine transaminase （ALT） and aspartate aminotransferase （AST） in serum were measured and liver histopathology was observed. The protein levels of nuclear factor（NF）-κB and acetylated histone H3 in liver tissue were detected by Western Blot. The hepatic mRNA levels of NF-κB, HDAC3, toll-like receptor 4 （TLR4） and TNF-α were measured by real-time PCR. ELISA was used to determine the TNF-α in serum. The results were analyzed with t test. ResultsThe serum levels of ALT in control group, model group and TSA group were （122.00±45.29）, （459.33±167.58） and （217.33±49.25） U/L, respectively. The differences between model group and control group or TSA group were significant （t=4.76 and 3.41, respectively, both P〈0.05）. The serum levels of AST in control group, model group and TSA group were （127.83±18.55）, （389.67±87.14） and （249.50±71.72） U/L, respectively. The differences between model group and control group or TSA group were also significant （t=7.20 and 3.04, respectively, both P〈0.05）. The inflammation of the liver histopathology induced by s100 was alleviated by TSA. The relative expressions of NF-κB protein, NF-κB mRNA, TNF-α mRNA, HDAC3 mRNA and TLR4 mRNA in the liver tissue of model group mice were 2.43±0.42, 9.51±0.36, 10.

关 键 词：组蛋白去乙酰化酶 曲古抑菌素A 核因子-ΚB 肿瘤坏死因子-α 肝炎 自身免疫性 

分 类 号：R575.1[医药卫生—消化系统]