基于自身免疫调节因子探讨左归丸对先天肾虚仔鼠T细胞迁移的影响  

Effects of Zuogui Pills on T cell migration based on AIRE in congenital kidney deficiency offspring rat

作　　者：任梅荣 曹继刚 桑红灵 徐安莉 萧闵[3] 陈会敏 赵敏 REN Mei-rong;CAO Ji-gang;SANG Hong-ling;XU An-li;XIAO Min;CHEN Hui-min;ZHAO Min(School of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Clinical College of Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Center Laboratory of Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学基础医学院,武汉430065 [2]湖北中医药大学中医临床学院,武汉430065 [3]湖北中医药大学中医药实验中心,武汉430065

出　　处：《中华中医药杂志》2021年第4期1934-1939,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81803969);第四批全国中医(临床、基础)优秀人才研修项目(No.国中医药人教发[2017]24);武汉市健康委员会(No.[2019]17)。

摘　　要：目的:观察左归丸对先天肾虚仔鼠胸腺和胸腺上皮细胞(TECs)内自身免疫调节因子(AIRE)、胰岛素样生长因子2(IGF2)、套膜蛋白(Involucrin)的表达和T细胞分类的影响。方法:构建先天肾虚仔鼠模型,各组孕鼠分别给予0.9%氯化钠溶液,胸腺肽,左归丸大、小剂量混悬液灌胃,至仔鼠出生。正常仔鼠胸腺TECs做原代培养,分为空白组、左归丸组、抑制剂组和抑制剂左归丸组。流式细胞学检测仔鼠胸腺和脾脏内T细胞分型,Western blot和RT-PCR检测胸腺和TECs内AIRE、IGF2、Involucrin蛋白和基因表达。结果:与对照组比较,模型组仔鼠脾脏内CD_4^(+)/CD_(8)^(+)T细胞,胸腺内AIRE、IGF2、Involucrin蛋白和基因表达均降低(P<0.05),胸腺内SP细胞增加(P<0.05);胸腺肽组及左归丸低、高剂量组仔鼠脾脏内CD_4^(+)/CD_(8)^(+)T细胞,胸腺内AIRE、IGF2、Involucrin蛋白和基因表达升高(P<0.05),胸腺内SP细胞减少(P<0.05)。与空白组比较,抑制剂组TECs内AIRE、IGF2、Involucrin蛋白和基因表达减少(P<0.05);与抑制剂组比较,左归丸组和抑制剂左归丸组TECs内AIRE、IGF2、Involucrin蛋白和基因的表达上升(P<0.05)。结论:先天肾虚仔鼠脾脏内CD_4^(+)/CD_(8)^(+)失衡,左归丸可以通过提高胸腺内AIRE、IGF2、Involucrin的表达,促进T细胞在胸腺正常成熟和外迁,纠正外周免疫器官脾脏的失衡状态。Objective:To observe the effects of Zuogui Pills(ZGP)on the expression of AIRE,IGF2,involucrin in thymus and TECs of offspring rats with congenital kidney deficiency(CKD)and the classification of T cells.Methods:Rats in each group were given normal saline,thymosin,ZGP high dose and low dose suspension respectively by gavage until their offspring were born.TECs of normal offspring were cultured and divided into blank group,ZGP group,inhibitor group,inhibitor-ZGP group.Flow cytometry was used to detect the T cell types in thymus and spleen of offspring rats.Western blot and RT-PCR were used to detect the protein and gene expression of AIRE,IGF2 and Involucrin in thymus and TECs.Results:Compared with the control group,CD_4^(+)/CD_(8)^(+)T cells in spleen,the protein and gene of AIRE,IGF2,Involucrin in thymus of model group offspring were decreased(P<0.05),SP cells in thymus were increased(P<0.05);The CD_4^(+)/CD_(8)^(+)T cells in spleen,the protein and gene of AIRE,IGF2,Involucrin in thymus were increased in thymosin group,ZGP low-dose group and ZGP high-dose group(P<0.05),SP cells in thymus were decreased(P<0.05).Compared with the blank group,the protein and gene of AIRE,IGF2 and Involucrin in the inhibitor group were decreased(P<0.05);compared with the inhibitor group,the protein and gene of AIRE,IGF2,Involucrin in TECs of ZGP group and inhibitor-ZGP group were increased(P<0.05).Conclusion:The CD_4^(+)/CD_(8)^(+)in spleen of offspring rats with CKD were unbalanced.ZGP can increase the expression of AIRE,IGF2 and Involucrin in thymus,promote the maturation and migration of T cells in thymus.Finally,the peripheral immune organ spleen imbalance can be restored.

关 键 词：先天肾虚 补肾填精法 左归丸 自身免疫调节因子 套膜蛋白 胰岛素样生长因子2 

分 类 号：R285.5[医药卫生—中药学]