【摘要】 【目的】研究高浓度雌激素(EE)对急性自身免疫性肝炎(AIH)的影响,并初步探索其可能的作用机制。【方法】通过小鼠尾静脉注射刀豆蛋白A,建立急性自身免疫性肝炎模型。将20只8周龄雌性C57BL/6小鼠按体质量随机区组分组,分为4组:对照(Control)组,刀豆蛋白A(ConA)组,雌二醇(EE)组和雌二醇+刀豆蛋白组(EE+ConA),每组5只。雌鼠卵巢去势,分别持续腹腔注射雌二醇2周,最后一次给药24 h后,尾静脉注射刀豆蛋白A。模型的评估采用苏木素-伊红染色法和血清ATL和AST测定法。TUNEL法检测肝细胞凋亡情况,酶联免疫吸附实验检测血清TNF-α和IL-6的含量。利用小鼠肝细胞系AML2,各处理组加入不同浓度的雌二醇,处理24 h和48 h,CCK8法检测各组细胞增殖情况,Annexin V-PI法检测各组细胞凋亡率。【结果】苏木素-伊红染色结果显示:分别与Control组和ConA组相比,EE+ConA组肝脏损伤程度加重,且血清ALT和AST水平明显增高,(P<0.000 1;P<0.000 1),差异具有统计学意义。TUNEL染色结果显示:分别与Control组和ConA组相比,EE+ConA组TUNEL阳性细胞数量明显增高,(P<0.000 1),差异具有统计学意义。酶联免疫吸附实验结果表明:分别与Control组和ConA组相比,EE+ConA组IL-6,TNF-α浓度明显增高,(P<0.000 1,P=0.024 7),差异具有统计学意义。CCK8检测细胞活性发现:高浓度雌二醇可以抑制AML12增殖活性,P<0.05,差异具有统计学意义。流式细胞结果显示:与Control组相比,高浓度雌二醇可以促进肝细胞凋亡,P<0.05,差异具有统计学意义。【结论】高浓度的雌二醇可能通过促进炎症因子TNF-α和IL-6的分泌,诱导肝细胞细胞凋亡,加重AIH小鼠肝脏炎症反应。更多还原

【Abstract】 【Objective】To study the effect of the high-concentration estrogen(EE)on acute autoimmune hepatitis(AIH)and to explore its possible molecular mechanism.【Methods】Twenty 8-week-old female C57BL/6 mice were randomly divided into 4 groups according to body weight,5 in each group. The mice in control(CON)group,ConA-treated(ConA)group,estradiol-treated(EE)group and treatment-combined(EE+ConA)group were treated with EE for two weeks by intraperitoneal injection after ovariectomy. After the last administration for 24 h,the caudal vein was injected with concanavalin A to establish AIH model. H&E,serum ATL,and AST were used to test AIH model. Apoptosis in liver tissues was detected by TUNEL. ELISA was employed to determine the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in serum. AML2 was treated with EE of different concentrations for 24 h and 48 h respectively. The cell viability was detected by CCK-8 assay and apoptosis was detected by Annexin V-FITC/PI flow cytometry.【Results】H&E and serum revealed that compared with those in Control and ConA,the degree of liver damage was increased significantly in EE+ConA group. The level of serum ALT and AST in EE+ConA group were also increased significantly(P < 0.000 1,P <0.000 1). TUNEL:The numbers of TUNEL positive cells increased significantly in EE+ConA,(P<0.000 1). ELISA results showed that compared with those in Control and ConA,in EE+ConA,the levels of IL-6 and TNF-α in serum had a significant rise,(P < 0.0001,P=0.024 7). Compared with Control and ConA,high concentration of EE treated AML12 group indicated lower cell viability,P < 0.05. In light of the result of flow cytometry,high concentration of EE could significantly promote hepatocyte apoptosis,(P<0.05).【Conclusion】High concentration of estradiol may promote the secretion of inflammatory cytokines such as TNF-α and IL-6,induce hepatocyte apoptosis,and aggravate the hepatic inflammation in AIH mice.更多还原