甲状腺滤泡细胞的自噬活性参与桥本甲状腺炎的发病机制研究  

Autophagy activity of thyroid follicular cells is involved in animal experiments on the pathogenesis of hashimoto's thyroiditis.

作　　者：马丁[1] 马昌军[1] 卢富琨 熊小芹[1] MA Ding;MA Chang-jun;LU Fu-kun(Department of Endocrinology,Bazhong Central Hospital,Bazhong Sichuan 636000,China.)

机构地区：[1]四川省巴中市中心医院内分泌科,四川巴中636000

出　　处：《临床和实验医学杂志》2021年第10期1025-1029,共5页Journal of Clinical and Experimental Medicine

基　　金：2018年四川省级科技计划项目(编号:2018JY0308)。

摘　　要：目的探讨甲状腺滤泡细胞的自噬活性参与桥本甲状腺炎(HT)的发病机制的动物实验。方法将30只NOD.H-2h4小鼠按照随机数字表法分为对照组和HT组,各15只。通过组织病理学观察小鼠中甲状腺炎的发生情况。通过实时聚合酶链式反应(RT-PCR)分析2组小鼠中促炎因子白细胞介素(IL)-23、IL-1β和干扰素-γ(IFN-γ)的mRNA表达。通过蛋白质印迹法分析小鼠LC3-II和Beclin及凋亡相关因子的蛋白表达。并通过免疫荧光法和流式细胞仪检测小鼠中活性氧(ROS)的积累。通过免疫组化分析AKT/mTOR/核因子κB(NF-κB)信号通路蛋白的表达。结果HT组小鼠甲状腺组织中单核细胞的浸润程度为(1.75±0.25)分,高于对照组[(0.67±0.12)分],差异有统计学意义(P<0.05)。HT组小鼠甲状腺组织中IL-23、IL-1β和IFN-γ的mRNA表达水平分别为2.37±0.42、3.04±0.71、2.85±0.35,均高于对照组(1.26±0.14、1.15±0.16、1.09±0.12),差异均有统计学意义(P<0.05)。HT组小鼠甲状腺组织中LC3-Ⅱ和Beclin的蛋白表达水平分别为1.25±0.18、1.17±0.15,均低于对照组(2.07±0.34、2.59±0.46),差异均有统计学意义(P<0.05)。HT组小鼠甲状腺组织中Caspase-3、Caspase-9、Bax的蛋白表达水平分别为2.16±0.34、2.74±0.46、2.51±0.38,均高于对照组(1.18±0.15、1.32±0.17、1.08±0.09),Bcl-2蛋白表达水平(1.07±0.12)低于对照组(2.14±0.32),差异均有统计学意义(P<0.05)。HT组小鼠甲状腺滤泡细胞中阳性细胞占比、免疫荧光强度值分别为(62.79±10.47)%、43.18±7.26,均高于对照组[(12.56±3.15)%、16.72±3.58],差异均有统计学意义(P<0.05)。HT组小鼠甲状腺组织中AKT、mTOR和NF-κB的蛋白表达水平分别为2.34±0.41、2.15±0.38、2.09±0.21,均高于对照组(1.15±0.10、1.23±0.14、1.08±0.12),差异均有统计学意义(P<0.05)。结论在HT发病机制中,炎症细胞因子IL-23、IL-1β和IFN-γ通过AKT/mTOR/NF-κB信号通路诱导甲状腺滤泡细胞自噬激活,这会导�Objective To investigate the autophagy activity of thyroid follicular cells involved in the pathogenesis of hashimoto's thyroiditis(HT)in animal experiments.Methods Thirty NOD.H-2h4 mice were divided into control group and HT group according the random number table method,15 mice in each group.The occurrence of thyroiditis in mice was observed by histopathology.The mRNA expression of pro-inflammatory factors IL-23,IL-1βand interferon-gamma(IFN-γ)in the two groups of mice was analyzed by real-time polymerase chain reaction.The protein expression of mouse LC3-II,Beclin and apoptosis-related factors was analyzed by Western blotting.The accumulation of ROS in mice was detected by immunofluorescence and flow cytometry.Analyze the expression of AKT/mTOR/NF-κB signaling pathway proteins by immunohistochemistry.Results The degree of infiltration of monocytes in the thyroid tissue of mice in the HT group was(1.75±0.25)points,which was higher than that in the control group[(0.67±0.12)points],and the difference was statistically significant(P<0.05).The mRNA expression levels of IL-23,IL-1βand IFN-γin the thyroid tissue of mice in the HT group were 2.37±0.42,3.04±0.71,2.85±0.35,which were higher than those in the control group(1.26±0.14,1.15±0.16,1.09±0.12),the differences were statistically significant(P<0.05).The protein expression levels of LC3-Ⅱand Beclin in the thyroid tissue of mice in the HT group were 1.25±0.18 and 1.17±0.15,respectively,which were lower than those of the control group(2.07±0.34,2.59±0.46),and the differences were statistically significant(P<0.05).The protein expression levels of Caspase-3,Caspase-9 and Bax in the thyroid tissue of mice in the HT group were 2.16±0.34,2.74±0.46,2.51±0.38,respectively,which were higher than those in the control group(1.18±0.15,1.32±0.17,1.08±0.09),the Bcl-2 protein expression level(1.07±0.12)was lower than that in the control group(2.14±0.32),the difference was statistically significant(P<0.05).The percentage of positive cells in the thyroid

关 键 词：小鼠 桥本甲状腺炎 甲状腺滤泡细胞 自噬 活性氧 炎症 

分 类 号：R581.4[医药卫生—内分泌]