Department of Cardiology, Beijing Anzhen Hospital, Beijing Institute of Heart, Lung and Blood Vessel Diseases, Capital Medical University, Beijing, China.
J Biochem Mol Toxicol. 2023 Sep;37(9):e23419. doi: 10.1002/jbt.23419. Epub 2023 Jun 21.
In the context of diabetes mellitus (DM), the circulating cathepsin S (CTSS) level is significantly higher in the cardiovascular disease group. Therefore, this study was designed to investigate the role of CTSS in restenosis following carotid injury in diabetic rats. To induce DM, 60 mg/kg of streptozotocin (STZ) in citrate buffer was injected intraperitoneally into Sprague-Dawley rats. After successful modeling of DM, wire injury of the rat carotid artery was performed, followed by adenovirus transduction. Levels of blood glucose and Th17 cell surface antigens including ROR-γt, IL-17A, IL-17F, IL-22, and IL-23 in perivascular adipose tissues (PVAT) were evaluated. For in vitro analysis, human dendritic cells (DCs) were treated with 5.6-25 mM glucose for 24 h. The morphology of DCs was observed using an optical microscope. CD4 T cells derived from human peripheral blood mononuclear cells were cocultured with DCs for 5 days. Levels of IL-6, CTSS, ROR-γt, IL-17A, IL-17F, IL-22 and IL-23 were measured. Flow cytometry was conducted to detect DC surface biomarkers (CD1a, CD83, and CD86) and Th17 cell differentiation. The collected DCs presented a treelike shape and were positive for CD1a, CD83, and CD86. Glucose impaired DC viability at the dose of 35 mM. Glucose treatment led to an increase in CTSS and IL-6 expression in DCs. Glucose-treated DCs promoted the differentiation of Th17 cells. CTSS depletion downregulated IL-6 expression and inhibited Th17 cell differentiation in vitro and in vivo. CTSS inhibition in DCs inhibits Th17 cell differentiation in PVAT tissues from diabetic rats following vascular injury.
在糖尿病（DM）的背景下，心血管疾病组的循环组织蛋白酶 S（CTSS）水平显著升高。因此，本研究旨在探讨 CTSS 在糖尿病大鼠颈动脉损伤后再狭窄中的作用。为了诱导 DM，将柠檬酸缓冲液中的 60mg/kg 链脲佐菌素（STZ）腹腔注射到 Sprague-Dawley 大鼠体内。成功建立 DM 模型后，对大鼠颈动脉进行钢丝损伤，然后进行腺病毒转导。评估血管周围脂肪组织（PVAT）中血糖水平和 Th17 细胞表面抗原（包括 ROR-γt、IL-17A、IL-17F、IL-22 和 IL-23）的水平。对于体外分析，用 5.6-25mM 葡萄糖处理人树突状细胞（DC）24h。用光学显微镜观察 DC 的形态。将来自人外周血单核细胞的 CD4 T 细胞与 DC 共培养 5 天。测量 IL-6、CTSS、ROR-γt、IL-17A、IL-17F、IL-22 和 IL-23 的水平。进行流式细胞术检测 DC 表面标志物（CD1a、CD83 和 CD86）和 Th17 细胞分化。收集的 DC 呈树突状，CD1a、CD83 和 CD86 阳性。葡萄糖在 35mM 剂量下损害 DC 的活力。葡萄糖处理导致 DC 中 CTSS 和 IL-6 的表达增加。葡萄糖处理的 DC 促进 Th17 细胞的分化。CTSS 耗竭下调体外和体内 DC 中 IL-6 的表达并抑制 Th17 细胞分化。CTSS 抑制 DC 可抑制糖尿病大鼠血管损伤后 PVAT 组织中的 Th17 细胞分化。
