树突状细胞组织蛋白酶S的下调抑制Th17细胞的分化以改善糖尿病血管损伤再狭窄  

Downregulation of cathepsin S in dendritic cells inhibits the differentiation of Th17 cells to ameliorate restenosis after vascular injury in diabetes

作　　者：李长江 彭红玉[1] 何松原 程子超 柳景华[1] Li Changjiang;Peng Hongyu;He Songyuan;Cheng Zichao;Liu Jinghua(Department of Cardiology,Beijing Anzhen Hospital,Capital Medical University,Coronary Heart Disease Center,Beijing 100029,China;Beijing Institute of Cardiopulmonary and Vascular Diseases,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院心内科冠心病中心,北京100029 [2]北京市心肺血管疾病研究所,北京100029

出　　处：《中华内分泌代谢杂志》2024年第8期681-689,共9页Chinese Journal of Endocrinology and Metabolism

基　　金：北京市属医院科研培育项目(PX2021026)。

摘　　要：目的探讨组织蛋白酶S(cathepsin S,CTSS)在糖尿病(diabetes mellitus,DM)诱导血管损伤再狭窄中的作用。方法(1)用不同浓度的葡萄糖刺激树突状细胞(dendritic cell,DC),通过腺病毒转染技术敲低和上调DC中的CTSS。用RT-qPCR和Western blot检测CTSS mRNA和蛋白表达水平,用RT-qPCR和ELISA检测CTSS敲低和上调时白细胞介素(IL)-6水平的变化。(2)用流式细胞术检测CTSS下调或过表达时Th17细胞分化程度,并检测ROR-γt、IL-17A、IL-17F、IL-22和IL-23的水平。(3)使用链脲佐菌素(streptozotocin,STZ,60 mg/kg)腹腔注射禁食12 h大鼠以获得DM大鼠模型,球囊导管与颈动脉导丝损伤方式获得再狭窄模型,上调和下调CTSS时比较不同组大鼠的Th17细胞分化程度。结果(1)35 mmol/L葡萄糖处理48 h的DC活力降低。与对照组相比,不同浓度葡萄糖处理DC中CTSS和IL-6水平呈现浓度依赖性升高(P<0.05),抑制DC中CTSS则降低IL-6蛋白水平,反之则升高(P<0.05)。(2)与对照组相比,在DC中抑制CTSS降低Th17细胞在T细胞中的百分比,ROR-γt、IL-17A、IL-17F、IL-22和IL-23的蛋白水平下调,反之则升高(P<0.05)。(3)与对照组相比,颈动脉损伤后大鼠血管周围脂肪组织(PVAT)中的CTSS表达增加,且PVAT组织中ROR-γt、IL-17A、IL-17F、IL-22和IL-23的水平也显著升高,而下调CTSS则消除葡萄糖诱导的增强效应。结论抑制DC中的CTSS导致Th17细胞分化程度降低,进而抑制糖尿病血管损伤再狭窄。Objective To explore the role of cathepsin S(CTSS)in diabetic vascular injury-induced restenosis.Methods(1)Dendritic cells(DCs)were stimulated with different concentrations of glucose,and CTSS was either knocked down or upregulated in dendritic cells using adenovirus transfection.The mRNA and protein expression levels of CTSS were detected by RT-qPCR and Western blot,and the changes of interleukin(IL)-6 levels were assessed using RT-qPCR and ELISA in response to CTSS.(2)The extent of Th17 cell differentiation was evaluated with Flow cytometry when CTSS was downregulated or overexpressed.Levels of ROR-γt,IL-17A,IL-17F,IL-22,and IL-23 were measured.(3)Streptozomycin(STZ,60 mg/kg)was injected into the intraperitoneal cavity of rats fasted for 12 h to obtain a diabetic rat model,and the restenosis model was obtained by balloon catheter and carotid guidewire injury,and the differentiation degree of Th17 cells in different groups of rats was compared when CTSS was up-regulated and down-regulated.Results(1)DC viability decreased when stimulated with 35 mmol/L glucose for 48 hours.Compared to the control group,glucose treatment led to a concentration-dependent increase in CTSS and IL-6 levels in DCs(P<0.05).Inhibition of CTSS reduced IL-6 protein levels,while its overexpression increased IL-6 protein levels(P<0.05).(2)Compared with the control group,CTSS inhibition in DC decreased the percentage of Th17 cells in T cells,with decreased protein levels of ROR-γt,IL-17A,IL-17F,IL-22,and IL-23,and vice versa(P<0.050).(3)After carotid artery injury,CTSS expression was increased in perivascular adipose tissue(PVAT)of rats,and levels of ROR-γt,IL-17A,IL-17F,IL-22,and IL-23 in PVAT were significantly elevated.Down-regulation of CTSS eliminated the glucose-induced enhancement.Conclusion Inhibition of CTSS in DC reduces Th17 cell differentiation and thereby suppresses restenosis following diabetic vascular injury.

关 键 词：糖尿病 组织蛋白酶S 树突状细胞 血管周围脂肪组织 TH17细胞 

分 类 号：R587.2[医药卫生—内分泌]