TNFR2与三氯乙烯致敏小鼠免疫性肝损伤的关系研究  

The relationship between TNFR2 and immune liver injury induced by trichloroethylene in mice

作　　者：杨呓 徐琼影 黄猛 姜伟 丁百旺 张家祥 朱启星 Yang Yi;Xu Qiongying;Huang Meng(Dept of Occupational and Environmental Health,School of Public Health,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学公共卫生学院职业卫生与环境卫生学系,合肥230032 [2]安徽医科大学第一附属医院皮肤病研究所,合肥230032

出　　处：《安徽医科大学学报》2021年第4期602-607,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81874259、81673141);安徽省自然科学基金(编号:2008085QH385)。

摘　　要：目的观察三氯乙烯(TCE)致敏小鼠肝脏内TNFR2表达情况,探讨TNFR2与免疫性肝脏损伤之间的关系。方法选取21只SPF级雌性BALB/c小鼠,6~8周龄,随机将其分为空白对照组(5只),溶剂对照组(5只),TCE处理组(11只),建立TCE致敏小鼠模型,进行小鼠背部皮肤致敏反应评分,评分大于1分为TCE致敏组,反之为未致敏组。模型完成后处死小鼠,摘取眼球取血,测定血清肝功能活性指标ALT和AST;取出小鼠肝脏制成石蜡切片,HE染色观察小鼠肝脏病理表现,免疫组织化学(IHC)法和Western blot法观察小鼠肝脏肿瘤坏死因子(TNF)-α与TNFR2的表达情况。实时荧光定量PCR法检测TNFR2的mRNA水平。结果TCE组致敏率为45.45%(5/11);致敏组小鼠的肝功能指标较高,与其他组比较差异有统计学意义(P<0.05)。HE染色结果显示TCE致敏组可见细胞发生空泡样变性,细胞呈蜂窝状且胞质疏松,其余组小鼠的肝细胞形态正常,染色均匀。免疫组化实验显示,TCE致敏组TNF-α表达较高,且与其他组相比TCE致敏组可以观察到大量TNFR2阳性表达,评分结果差异有统计学意义(P<0.05)。实时荧光PCR结果显示TCE致敏组TNFR2的mRNA表达也比其他组表达升高(P<0.05)。Western blot实验表明TCE致敏组比其他组TNFR2蛋白表达水平增高,差异有统计学意义(P<0.05)。结论TNFR2在TCE致敏组小鼠肝脏中表达升高,其表达的改变可能参与了TCE致敏小鼠免疫性肝脏损伤过程。Objective To observe the expression of TNFR2 in the liver of mice and to explore the relationship between TNFR2 and the immune liver injury induced by trichloroethylene in mice.Methods Twenty-one BALB/c mice from 6 to 8 weeks of age were randomly divided into blank control group(n=5),solvent control group(n=5)and TCE treatment group(n=11).A TCE-sensitized mouse model was established according to the pre-sensitization method of the research group.Twenty-four hours after the last challenge,the skin sensitization response of the mice was scored,and the mice in the TCE group were divided into the TCE-sensitized group and the TCE non-sensitized group.The mice were sacrificed 72 hours after the last challenge,and the blood was collected from the venous plexus of the eye to measure liver function indexes such as alanine aminotransferase(ALT)and aspartate aminotransferase(AST).The liver of the mice was taken out and made into paraffin sections.HE staining was used to observe the pathological features of the liver.Immunohistochemistry method and Western blot method were used to determine the expression of TNF-αand TNFR2 in mouse liver.Real-time fluorescence quantitative PCR method was used to detect the level of TNFR2 gene in mouse liver.Results The sensitization rate of the TCE group was 45.45%(5/11);the serum ALT and AST levels of the TCE-sensitized group were significantly higher than those of the blank control group,solvent control group and TCE unsensitized group,and the difference was statistically significant(P<0.05).HE staining results showed that the blank control group,the solvent control group and the TCE unsensitized group had normal liver cell morphology and even staining.The TCE-sensitized group showed cell swelling and vacuolar degeneration.Immunohistochemical experiments showed that the TCE-sensitized group had a higher expression of TNF-α,a large number of TNFR2 positive expressions could be observed in the TCE-sensitized group compared with other groups,and the difference in scores was statistically si

关 键 词：三氯乙烯 肝脏 TNFR2 职业性三氯乙烯药疹样皮炎 

分 类 号：R135.7[医药卫生—劳动卫生]