摘要
目的探讨雷帕霉素对刀豆蛋白A（ConA）诱导的自身免疫性肝炎及纤维化进展的防治作用，并探讨其机制。方法8周龄雌性C57BL／6小鼠随机分为正常对照组、ConA造模组、ConA造模＋雷帕霉索（RAPA）治疗组。检测各组小鼠血清丙氨酸氨基转移酶（ALIT）和天冬氨酸氨基转移酶（AST）水平；利用苏木素-伊红和Masson染色方法并行KnodellHAI和Ishak肝纤维化半定量计分评价肝脏组织炎症及纤维化程度；梯度离心法分离肝脏单个核细胞，流式细胞术检测CD4＋T、CD8＋T细胞的比例及细胞内因子染色分析免疫细胞内干扰素Y（IFNY）、白细胞介素4（IL-4）、白细胞介素10（IL-10）、转化生长因子p（TGFp）的表达情况。组间数据比较用f检验。结果雷帕霉索治疗组与ConA造模组小鼠相比，血清ALT水平显著降低妒〈0．05）；肝脏组织炎症损伤明显减轻，并无明显纤维组织增生；肝脏单个核细胞内TGFp的表达显著降低[治疗组（8．91％±1．25％）对比造模组（16．65％±2．05％），P〈0．05）；肝脏CD4＋T和CD80细胞的比例均下调[治疗组（4．09％±1．20％、3．28％±0．66％）对比造模组（8．91％±0．69％、9．68％±1．46％），P〈0．05）；Th1细胞的比例降低[治疗组（1．02％±0．06％）对比造模组（2．83％±0．21％），P〈0．05]，Th3、Tr1调节性T细胞的比例显著上调[治疗组（59．53％±9．8砒、10．63％±2．27％）对比造模组（47．13％±4．79％、7．09％±1．66％），P〈0．05]，但Th2细胞的差异无统计学意义（P〉0．05）。结论雷帕霉索可能通过促进肝脏Th3／Tr1细胞的分化，减少肝脏单个核细胞TGFD的表达，减缓ConA诱导的慢性肝炎肝纤维化的进展，从而具有预防肝纤维化的作用。
Objective To investigate the preventive and therapeutic effects of rapamycin （RAPA） on autoimmune hepatitis and liver fibrosis induced by concanavalin A （ConA） and possible mechanisms. Methods Female C57BL/6 mice aged 8 weeks were randomly divided into normal control group, CortA model group, and ConA＋RAPA treatment group. The serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were measured; hematoxylin-eosin and Masson staining and Knodell HAI and Ishak scoring systems were used to evaluate the degrees of liver inflammation and fibrosis. Gradient centrifugation was used to separate mononuclear cells, flow cytometry was used to measure CD4＋/CD8＋ ratio, and intracellular cytokine staining was performed to measure the levels of interferon-7 （IFN-？）, interleukin-4 （/L-4）, interleukin-10 （IL-10）, and transforming growth factor 13 （TGF-13） in immune cells. The t-test was used for data comparison between groups. Results The RAPA treatment group showed a significant reduction in serum ALT level compared with the ConA model group （P 〈 0.05）. Liver inflammatory injury was reduced significantly, and there was no obvious fibrous tissue proliferation. The level of TGF-13 in mononuclear cells was reduced significantly, and the treatment group had a significantly lower level of TGF-13 than the model group （8.91%±1.25% vs 16.65%±2.05%, P 〈 0.05）. The proportions of CD4＋ and CD8＋T cells in the liver were reduced, and the treatment group had significantly lower proportions of CD4＋ and CD8＋T cells than the model group （proportion of CD4＋T cells： 4.09%±1.20% vs 8.91%±0.69%, P 〈 0.05; proportion of CD8＋T cells： 3.28%±0.66% vs 9.68%±1.46%, P 〈 0.05）. The proportion of Thl cells was reduced, and the treatment group had a significantly lower proportion of Thl cells than the model group （1.02%±0.06% vs 2.83%±0.21%,P 〈 0.05）; the proportions of Th3 and Trl regulatory T cells were increased, and the treatment group had significantly higher proportions of Th3 and Trl regulatory T cells than the model group （proportion of Th3 regulatory T cells： 59.53%＋9.82% vs 47.13%±4.79%, P 〈 0.05; proportion of Trl regulatory T cells： 10.63%±2.27% vs 7.09%±l.66%, P 〈 0.05）, but the proportion of Th2 cells showed no significant difference between the two groups （P 〉 0.05）. Conclusion RAPA can promote the differentiation of Th3/Trl cells, reduce the expression of TGF-β in mononuclear cells, slow down the progression of chronic hepatitis induced by ConA into liver fibrosis, and thus prevent liver fibrosis.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2016年第5期368-374,共7页
Chinese Journal of Hepatology
基金
中国肝炎防治基金会-天晴肝病研究基金资助课题，国家自然科学基金，天津市卫生局科技攻关项目(12KG133)China Foundation for Hepatitis Prevention and Control TianQing Liver Disease Funds，National Natural Science Foundation of China，Science and Technology Project of Tianjin Health and Family Planning Commision
