摘要
目的探讨白芍总苷对BALB/c小鼠Graves病(GD)模型的保护作用及其机制。方法将50只(6周龄,体重16~18 g)无特定病原体级雌性BALB/c小鼠按随机数字表法分为5组,分别设为对照组、模型组、早期低剂量白芍总苷干预组(250 mg·kg^(-1)·d^(-1))、早期高剂量白芍总苷干预组(500 mg·kg^(-1)·d^(-1))和晚期白芍总苷干预组,每组各10只。除对照组外,其余4组均注射表达促甲状腺激素受体(TSHR)A亚单位的重组腺病毒进行3次免疫(第0、3和6周),完成GD建模;早期低剂量和高剂量干预组全程予以含不同剂量白芍总苷的饲料,晚期干预组自第2次免疫后1周(第4周)起加用含低剂量白芍总苷的饲料。于第4周取小鼠尾静脉血检测促甲状腺素受体抗体(TRAb)、总甲状腺素(TT4)水平;第10周检测小鼠血清TRAb及TT4水平及各组调节性T细胞(Treg)比率,并观察甲状腺组织病理变化。检测各组血清辅助性T细胞1(Th1)及Th2细胞相关因子白细胞介素-2(IL-2)、IL-4、IL-5、IL-10、IL-12p70、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、干扰素-γ(IFN-γ)和肿瘤坏死因子-α(TNF-α)等水平,探讨白芍总苷对BALB/c小鼠GD模型的保护作用及其机制。结果第4周时,早期高剂量干预组TT4[(55.07±12.89)μg/L]水平低于模型组[(74.33±8.63)μg/L](P<0.05),早期低剂量干预组和晚期干预组TT4水平和模型组差异无统计学意义(均P>0.05);早期低剂量干预组、早期高剂量干预组和晚期干预组小鼠TRAb水平和模型组差异均无统计学意义(均P>0.05)。第10周时,早期高剂量干预组TRAb[(90.00±26.89)U/L]、TT4[(32.66±8.11)μg/L]水平均低于模型组[(396.97±95.35)U/L,(73.70±16.33)μg/L](均P<0.05),而早期低剂量干预组和晚期干预组小鼠TRAb、TT4水平与模型组差异均无统计学意义(均P>0.05)。早期高剂量干预组甲亢小鼠甲状腺组织呈局灶性增生性改变,而其他组甲亢小鼠甲状腺组织呈弥漫性增生性改变。第10周(3次重组腺病毒免疫后4周),早期高剂量干预组小鼠CD4^(+)CD25^(+)/CD4^(+)Treg比值高于模型组(P<0.05)。第10周小鼠血清细胞因子和模型组相比,早期高剂量干预组IL-2、IL-12p70和IFN-γ水平均降低(均P<0.05),IL-10水平升高(P<0.05)。结论早期高剂量(500 mg·kg^(-1)·d^(-1))白芍总苷对小鼠GD模型具有保护性作用,其作用机制可能和调节性T细胞功能改变及Th1/Th2细胞因子平衡恢复有关。
Objective To investigate the protective effect and its immunoregulatory mechanism of Total Glucosides of Paeony(TGP)against Graves′Disease(GD)model on BALB/c mice.Methods Fifty female(6 weeks old,weighing 16-18 g)BALB/c mice of specific pathogen free were divided into control group according to random number table method,model group,early low-dose TGP intervention group(250 mg·kg^(-1)·d^(-1)),early high-dose TGP intervention group(500 mg·kg^(-1)·d^(-1)),and late TGP intervention group,with 10 mice in each group.Except the control group,the other 4 groups were immunized 3 times(0,3^(rd),and 6^(th) week)with recombinant adenovirus expressing the thyroid stimulating hormone receptor(TSHR)A subunit to establish the GD model.The early low-dose and high-dose intervention group were given diets containing different doses of TGP throughout the whole process,and the late intervention group was given diets containing low doses of TGP from the 1^(st) week after the 2^(nd) immunization(week 4).The levels of thyrotropin receptor antibody(TRAb)and total thyroxine(TT4)were detected in the tail venous blood of mice at the 4^(th) week.At the 10^(th) week,the serum TRAb and TT4 levels and the ratio of regulatory T cells(Treg)in each group were detected,and the pathological changes of thyroid tissue were observed.Serum helper T cell 1(Th1)and Th2 cell-related factors interleukin-2(IL-2),IL-4,IL-5,IL-10,IL-12p70,granulocyte-macrophage colony-stimulating factor(GM-CSF),interferon-γ(IFN-γ)and tumor necrosis factors-α(TNF-α)were detected to investigate the protective effect of TGP on GD model in BALB/c mice and its mechanism.Results At the 4^(th) week,The level of TT4[(55.07±12.89)μg/L]in early high-dose intervention group was lower than that in model group[(74.33±8.63)μg/L](all P<0.05).The level of TT4 in early low-dose intervention group and late intervention group and model group had no statistical significance(all P>0.05).TRAb level of mice between early low-dose,early high-dose,late intervention groups and model group was no significant difference(all P>0.05).At the 10^(th) week,TRAb[(90.00±26.89)U/L]and TT4[(32.66±8.11)μg/L]levels in the early high-dose intervention group were lower than those in the model group[(396.97±95.35)U/L,(73.70±16.33)μg/L](all P<0.05).The TRAb and TT4 levels in the early low-dose intervention group and late intervention group were not significantly different from those in the model group(all P>0.05).The thyroid tissue of hyperthyroidism mice in the early high dose intervention group showed focal hypertrophic changes,while the thyroid tissue of other hyperthyroidism mice showed diffuse hypertrophic changes.The CD4^(+)CD25^(+)/CD4^(+)Treg ratio in early high-dose intervention group was higher than that in model group at the 10^(th) week(4 weeks after three recombinant adenovirus immunization)(P<0.05).Compared with the model group at the 10^(th) week,the levels of IL-2,IL-12p70 and IFN-γin the early high-dose intervention group were all decreased(all P<0.05),and the levels of IL-10 were increased(P<0.05).Conclusion Early high-dose(500 mg·kg^(-1)·d^(-1))TGP intervention group displays a protective effect against GD mice,the mechanism of which may be related to regulatory T cell function changes and Th1/Th2 cytokine balance restoration.
作者
伍丽萍
谢宝松
王静雅
德吉措穆
赵凤仪
丁习
刘星君
林晓瑛
周星辰
赵媛
吴谦
施秉银
Wu Liping;Xie Baosong;Wang jingya;Cuomu DeJi;Zhao Fengyi;Ding Xi;Liu Xingjun;Lin Xiaoying;Zhou Xingchen;Zhao Yuan;Wu Qian;Shi Bingyin(Department of Endocrinology,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China;Department of Gerontology,Shanxi Provincial People′s Hospital,Xi′an 710068,China;Department of Epidemiology,School of Public Health,Xi′an Jiaotong University Health Science Center,Xi′an 710061,China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2023年第9期689-695,共7页
National Medical Journal of China
基金
国家自然科学基金(81873638、81970679、82170805)
陕西省创新能力支撑计划(临床研究中心)(2021LCZX-05)
陕西省自然科学基础研究计划(2022 JM-586)
关键词
格雷夫斯病
动物模型
白芍总苷
细胞因子
免疫
Graves′disease
Animal model
Total glucosides of paeony
Cytokine
Immunization
作者简介
通信作者:施秉银,Email:shibingy@126.com
