Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, School of Pharmacy, Anhui Medical University, 81 Meishan Road, Hefei 230032, Anhui Province, PR China.
Department of Pathology, School of Basic Medicine, Anhui Medical University, 81 Meishan Road, Hefei 230032, Anhui Province, PR China.
Int Immunopharmacol. 2021 Jan;90:107214. doi: 10.1016/j.intimp.2020.107214. Epub 2020 Dec 2.
We previously revealed that the overexpression of synovial aquaporin 1 (AQP1) aggravated collagen-induced arthritis (CIA) in rats via regulating β-catenin signaling. This study was to demonstrate the therapeutic effect of acetazolamide (AZ, an AQP1 inhibitor) on rat CIA and explored its underlying mechanisms. Paw swelling, arthritis index, pathological assessments, and serum levels of collagen type II (Col II) antibody, IL-1β and TNF-α were measured to evaluate the anti-arthritic effect of AZ on rat CIA. Ki67 immunohistochemistry and TUNEL assay were performed to reveal the anti-proliferative and pro-apoptotic effects of AZ on synovial cells in vivo. The protein levels of apoptosis-related genes and Wnt/β-catenin pathway key members were detected by western blot. We found that AZ treatment on CIA rats could inhibit paw swelling, reduce arthritis index, alleviate the pathologic changes of ankle joint and decrease the serum levels of Col II antibody, TNF-α and IL-1β. AZ could reduce Ki67 expression and increase apoptosis index in CIA synovial tissues by reducing Bcl-2 protein level, increasing Bax and caspase 3 protein levels and normalizing Bcl-2/Bax ratio. Moreover, AZ could reduce the protein levels of Wnt1, β-catenin, p-GSK-3β (Ser9), c-myc, cyclin D1 and MMP9, while increase GSK-3β protein level in CIA synovial tissues. Importantly, these mentioned effects of AZ (60 mg/kg) on CIA rats could be reversed by the combined use of lithium chloride (LiCl), an activator of Wnt/β-catenin pathway. In short, AZ exerted potent anti-arthritic effects on CIA rats by inducing synovial apoptosis and inhibiting Wnt/β-catenin pathway.
我们之前的研究表明，滑液水通道蛋白 1（AQP1）的过表达通过调节β-连环蛋白信号加重了大鼠胶原诱导性关节炎（CIA）。本研究旨在探讨乙酰唑胺（AZ，AQP1 抑制剂）对大鼠 CIA 的治疗作用及其潜在机制。通过测量爪肿胀、关节炎指数、病理评估以及血清中 II 型胶原（Col II）抗体、IL-1β 和 TNF-α 的水平来评估 AZ 对 CIA 大鼠的抗关节炎作用。通过 Ki67 免疫组化和 TUNEL 检测来揭示 AZ 对体内滑膜细胞的抗增殖和促凋亡作用。通过 Western blot 检测凋亡相关基因和 Wnt/β-连环蛋白通路关键成员的蛋白水平。我们发现，AZ 治疗 CIA 大鼠可抑制爪肿胀，降低关节炎指数，减轻踝关节病理变化，降低血清 Col II 抗体、TNF-α 和 IL-1β 水平。AZ 可通过降低 Bcl-2 蛋白水平、增加 Bax 和 caspase 3 蛋白水平以及使 Bcl-2/Bax 比值正常化来减少 CIA 滑膜组织中的 Ki67 表达并增加凋亡指数。此外，AZ 可降低 CIA 滑膜组织中的 Wnt1、β-连环蛋白、p-GSK-3β（Ser9）、c-myc、细胞周期蛋白 D1 和 MMP9 蛋白水平，同时增加 GSK-3β 蛋白水平。重要的是，通过使用 Wnt/β-连环蛋白通路的激活剂氯化锂（LiCl），可以逆转 AZ（60mg/kg）对 CIA 大鼠的上述作用。总之，AZ 通过诱导滑膜细胞凋亡和抑制 Wnt/β-连环蛋白通路对 CIA 大鼠发挥了强大的抗关节炎作用。
