Notch1通路调控CD4^(+)CD25^(+)Foxp3^(+)调节性T细胞在肾血管性高血压中的作用  

The role of Notch1 signaling pathway regulating CD4^(+)CD25^(+)Foxp3^(+) regulatory T cells in renovascular hypertension

作　　者：余德宪[1] 郑娜[1] 尹丽娟[1] 陈旸[2] 彭琳 何发静[2] 陈文娟 YU Dexian;ZHENG Na;YIN Lijuan;CHEN Yang;PENG Lin;HE Fajing;CHEN Wenjuan(Department of Nephrology,Sichuan Academy of Medical Sciences,Sichuan Provincial People's Hospital,Chengdu,610072,China;Department of Cardiology,Sichuan Academy of Medical Sciences,Sichuan Provincial People's Hospital,Chengdu,610072,China)

机构地区：[1]四川省医学科学院·四川省人民医院肾内科,四川成都610072 [2]四川省医学科学院·四川省人民医院心内科,四川成都610072

出　　处：《中华高血压杂志（中英文）》2024年第4期363-370,共8页Chinese Journal of Hypertension

基　　金：成都市金牛区医学会2020年肾脏内科专委会专项科研课题(JNQN20-18)。

摘　　要：目的 探索Notch1通路调控CD4^(+)CD25^(+)Foxp3^(+)调节性T细胞(Treg)在肾血管性高血压(RVH)中的作用与机制。方法 选择2019年1月至2022年1月在四川省人民医院肾内科诊治为RVH的患者220例(RVH组),以及同期普通非RVH原发性高血压患者220例(原发性高血压组),另选择健康体检者220人为对照组,经倾向性评分匹配后,每组纳入10例。采用流式细胞术测定外周血单个核细胞(PBMC)中CD4^(+)CD25^(+)Foxp3^(+) Treg比例、白细胞介素-10(IL-10)、转化生长因子β(TGF-β)和细胞增殖抗原Ki-67水平;采用实时荧光定量PCR测定Notch1、Notch配体δ样配体(DDL)1、DDL4、Jagged1、Jagged4的mRNA水平;采用Western-blot法测定Notch1、hes家族bHLH转录因子(Hes)1、Hes5蛋白水平。结果 与对照组比较,RVH组PBMC中CD4^(+)CD25^(+)Foxp3^(+)Treg比例降低,TGF-β、Ki-67表达水平明显降低,IL-10表达水平升高(均P<0.001);与RVH组比较,原发性高血压组PBMC中CD4^(+)CD25^(+)Foxp3^(+)Treg比例升高,TGF-β和Ki-67表达水平升高,IL-10表达水平降低(均P<0.001)。实时定量PCR结果显示,RVH组患者PBMC中Notch受体Notch1 mRNA、DLL1、DLL4、Jagged1和Jagged2 mRNA相对表达量也高于对照组和原发性高血压组(P<0.001)。Western-blot结果显示,患者外周CD4^(+)CD25^(+)Foxp3^(+)Treg加入Notch1信号通路抑制剂后,Hes1与Hes5表达下降,CD4^(+)CD25^(+)Foxp3^(+)Treg占比和TGF-β表达水平上升(均P<0.001)。结论 Notch1信号通路在CD4^(+)CD25^(+)Foxp3^(+)Treg调节的RVH中起着免疫调控作用,调节Notch1信号通路可明显促进CD4^(+)CD25^(+)Foxp3^(+)Treg的增殖和分泌抑炎因子的能力。Objective To explore the role and mechanism of Notch1 pathway in regulating CD4^(+)CD25^(+) Foxp3^(+) regulatory T cells(Treg) in renal vascular hypertension(RVH). Methods Two hundred and twenty patients treated in the Department of Nephrology of Sichuan Provincial People's Hospital from January 2019 to January 2022 were enrolled as RVH group, and 220 patients with essential hypertension(EH) in the same period were selected as EH group;another 220 healthy examinees were selected as control group(NC). After propensity score matching, ten cases were included in each group.Flow cytometry was used to determine the proportion of CD4^(+)CD25^(+)Foxp3^(+) Treg, interleukin-10(IL-10), transforming growth factor-β(TGF-β), and Ki-67 levels in peripheral blood mononuclear cell(PBMC). Real-time quantitative polymerase chain reaction(PCR) was employed to measure the messenger RNA levels of Notch1, DLL, DLL4, Jagged1, and Jagged4. Western blot analysis was conducted to determine the protein levels of Notch1, hes family bHLH transcription factor(Hes)1, and Hes5. Results Flow cytometry results showed that compared with NC group, RVH group had significantly lower proportion of CD4^(+)CD25^(+)Foxp3^(+) Treg cells, TGF-β and Ki-67 expression levels, and higher IL-10 expression level, with significant differences(P<0.001). Compared with RVH group, EH group had significantly higher proportion of CD4^(+)CD25^(+)Foxp3^(+) Treg cells, TGF-β and Ki-67 expression levels, and lower IL-10 expression level, with significant differences(P<0.001). Real-time quantitative PCR results showed that RVH group also had significantly higher relative expression levels of Notch receptor Notch1 mRNA, Notch ligand δ-like ligand(DLL)1, DLL4, Jagged1 and Jagged2 mRNA in PBMC than NC group and EH group, with significant differences(P<0.001). Western blot results showed that after adding Notch1 signaling pathway inhibitor, Hes1 and Hes5 expression in peripheral CD4^(+)CD25^(+)Foxp3^(+) Treg cells of patients decreased significantly(P<0.001), while CD4^(+

关 键 词：肾血管性高血压 CD4^(+)CD25^(+)Foxp3^(+)调节性T细胞 Notch1通路 转化生长因子-β 

分 类 号：R544.14[医药卫生—心血管疾病]