原发性胆汁性胆管炎伴抑郁小鼠的肝肠损伤及肠黏膜中ZO-1的表达变化

Hepatointestinal injury and changes of ZO-1 expression in intestinal mucosa of mice with primary biliary cholangitis and depression

目的研究原发性胆汁性胆管炎(primary biliary cholangitis,PBC)伴抑郁小鼠的肝肠损伤及肠黏膜中闭锁小带蛋白1(zonula occludens protein 1,ZO-1)的表达变化。方法选取24只6周龄雌性C57BL/6小鼠,随机分为对照组、PBC组、PBC+抑郁组、PBC+抑郁+腹腔注射氯米帕明组,每组6只,根据不同实验组模型要求造模:抑郁模型小鼠采用慢性不可预知的轻度刺激方式处理,PBC组小鼠给予含0.1%3,5-二乙氧基羰基-1,4-二氢-2,4,6-三甲基吡啶(3,5-diethyloxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine,DDC)饲料喂养,PBC+抑郁+腹腔注射氯米帕明组小鼠每日同一时间腹腔内注射氯米帕明7.5mg/kg以诱导模型。小鼠完成造模后测量小鼠体质量分析造模情况。处死并同时采集肝组织及肠组织标本,苏木精-伊红(hematoxylin and eosin,HE)染色观察各组肝及肠组织的显微病理变化。应用免疫组织化学染色技术检测各组小鼠肠组织中ZO-1的表达水平。结果与对照组比较,各模型组小鼠的体质量均明显下降(P<0.05),PBC+抑郁组小鼠的体质量较PBC组小鼠降低(P<0.05),PBC+抑郁+腹腔注射氯米帕明组小鼠的体质量较PBC+抑郁组小鼠显著增高(P<0.05)。与对照组比较,各模型组小鼠的肝组织损伤严重,且出现不同程度的肠组织病理损伤,ZO-1在各模型组小鼠肠黏膜中的表达水平均显著下降(P<0.01);与PBC组比较,PBC+抑郁组小鼠的肠黏膜中ZO-1表达水平显著降低(P<0.01);与PBC+抑郁组比较,PBC+抑郁+腹腔注射氯米帕明组小鼠的肠黏膜中ZO-1表达水平显著升高(P<0.01)。结论抗抑郁治疗可显著改善PBC伴抑郁小鼠的肝肠损伤及肠黏膜中ZO-1的表达。

Objective To explore the hepatointestinal injury and changes of zonula occludens protein 1(ZO-1)expression in intestinal mucosa of mice with primary biliary cholangitis(PBC)and depression.Methods A total of 24 six-week-old female C57BL/6 mice were randomly divided into control group,PBC group,PBC+depression group,PBC+depression+intraperitoneal injection of clomipramine group,6 mice in each group.The models were made according to the requirements of different experimental groups.The depression model was treated with chronic unpredictable mild stimulation.PBC group was fed with feed containing 0.1%3,5-diethyloxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine(DDC).In the intraperitoneal injection group,7.5 mg/kg of clomipramine was injected intraperitoneally at the same time every day to induce the model.The body weight of the mice was measured to analyze the establishment of the model.The mice were sacrificed and liver and intestinal tissue samples were collected at the same time.Observation of histopathological changes of liver and intestinal tissues in each group by hematoxylin and eosin(HE)staining.Immunohistochemical staining was used to detect the expression level of ZO-1 in intestinal tissues of each group.Results Compared with control group,the body weight of the mice in each model group decreased significantly(P<0.05),and the body weight of PBC+depression group was lower than that of PBC group(P<0.05).The body weight of PBC+depression+intraperitoneal injection of clomipramine group was significantly higher than that of PBC+depression group(P<0.05).Compared with control group,the liver tissue of each model group was seriously injured,and there were pathological injuries of intestinal tissue with different severity.Compared with control group,the expression level of ZO-1 in the intestinal mucosa of each model group was significantly decreased(P<0.01).Compared with PBC group,the expression level of ZO-1 in PBC+depression group was significantly decreased(P<0.01).Compared with PBC+depression group,the expression level of ZO-1 in PBC+depression+intraperitoneal injection of clomipramine group was significantly increased(P<0.01).Conclusion Antidepressant treatment can significantly improve the hepatointestinal injury and the expression of ZO-1 in the intestinal mucosa of PBC with depression in mice.