Department of Anesthesiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.
Department of Anesthesiology, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, P.R. China.
Mol Med Rep. 2021 Jul;24(1). doi: 10.3892/mmr.2021.12180. Epub 2021 Jun 3.
Increasing evidence has demonstrated that regulatory T cells (Tregs) suppress innate immunity, as well as protect the kidneys from ischemia‑reperfusion injury (IRI) and offer a potentially effective strategy to prevent or alleviate renal IRI. The present study explored whether C‑X‑C motif chemokine receptor 3 (CXCR3) alleviated renal IRI by increasing Tregs. Male C57BL/6J mice were divided into sham‑surgery, IRI, CXCR3 overexpression (OE‑CXCR3)+IRI, PC61+IRI and OE‑CXCR3+PC61+IRI groups. Histopathological examination of the kidney was carried out using hematoxylin‑eosin and Masson staining. The levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were measured. Blood and kidney levels of IL‑6, TNF‑α, C‑C motif chemokine ligand (CCL)‑2 and IL‑10 were detected by ELISA and western blotting. The levels of superoxide dismutase (SOD), glutathione peroxidase (GSH‑Px) and malondialdehyde (MDA) in kidney tissues were also measured to assess oxidative stress. The population of Tregs in the kidney was assessed using flow cytometry. The results demonstrated that administration of OE‑CXCR3 to IRI mice significantly decreased the levels of Scr, BUN, IL‑6, TNF‑α, CCL‑2 and MDA, increased the levels of IL‑10, SOD and GSH‑Px, and mitigated the morphologic injury and fibrosis induced by IR compared with the IRI group. In addition, administration of OE‑CXCR3 induced significant reductions in the expression levels of fibrosis‑related markers, including fibronectin and type IV collagen, and increased the number of Tregs. These roles of OE‑CXCR3 were significantly neutralized following deletion of Tregs with PC61 (anti‑CD25 antibody). Together, the present study demonstrated that injection of OE‑CXCR3 lentiviral vectors into animal models can alleviate renal IRI by increasing the number of Tregs. The results may be a promising approach for the treatment of renal IRI.
越来越多的证据表明，调节性 T 细胞（Tregs）抑制固有免疫，保护肾脏免受缺血再灌注损伤（IRI），并提供一种预防或减轻肾 IRI 的潜在有效策略。本研究探讨了 C-X-C 基序趋化因子受体 3（CXCR3）是否通过增加 Tregs 来减轻肾 IRI。雄性 C57BL/6J 小鼠分为假手术、IRI、CXCR3 过表达（OE-CXCR3）+IRI、PC61+IRI 和 OE-CXCR3+PC61+IRI 组。采用苏木精-伊红和 Masson 染色对肾脏进行组织病理学检查。通过酶联免疫吸附试验（ELISA）和蛋白质印迹法检测血清肌酐（Scr）和血尿素氮（BUN）水平。通过 ELISA 和蛋白质印迹法检测血液和肾脏中白细胞介素 6（IL-6）、肿瘤坏死因子-α（TNF-α）、C 型趋化因子配体（CCL）-2 和白细胞介素 10（IL-10）的水平。还测量了肾组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和丙二醛（MDA）的水平，以评估氧化应激。通过流式细胞术评估肾脏中 Tregs 的群体。结果表明，与 IRI 组相比，OE-CXCR3 给药可显著降低 Scr、BUN、IL-6、TNF-α、CCL-2 和 MDA 的水平，增加 IL-10、SOD 和 GSH-Px 的水平，并减轻 IRI 引起的形态损伤和纤维化。此外，OE-CXCR3 给药可显著降低纤维化相关标志物（包括纤维连接蛋白和 IV 型胶原）的表达水平，并增加 Tregs 的数量。用 PC61（抗 CD25 抗体）去除 Tregs 后，OE-CXCR3 的这些作用明显被中和。综上所述，本研究表明，向动物模型注射 OE-CXCR3 慢病毒载体可通过增加 Tregs 的数量来减轻肾 IRI。这些结果可能为治疗肾 IRI 提供一种有前途的方法。
