The Department of Pharmacy, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, China; Anhui Province Key Laboratory of Major Autoimmune Diseases, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Hefei, 230032, China; The Key Laboratory of Anti-inflammatory and Immune Medicines, Ministry of Education, Hefei, 230032, China.
Institute of Clinical Pharmacology, Anhui Medical University, Hefei, 230032, China.
Eur J Pharmacol. 2019 Sep 15;859:172558. doi: 10.1016/j.ejphar.2019.172558. Epub 2019 Jul 17.
Proline-serine-threonine-phosphatase-interacting protein 2 (PSTPIP2) is related to inflammation. In this study, we investigated the function of PSTPIP2 in adjuvant-induced arthritis (AIA) by using adeno-associated virus (AAV) to overexpress PSTPIP2 in rat. AIA rats were developed by injecting Lewis rats with complete Freund's adjuvant (CFA) on day 0. AAV-empty or AAV-PSTPIP2, or PBS was administered intraarticularly into each knee joint on day 8 postinduction. All animals were killed at day 18 after adjuvant injection. WB was used to detect the expression of PSTPIP2 in rat synovial tissues. Fluorescence microscopy showed the transduction efficiency in synovial tissue. The morphology of arthritic joints was examined by HE, safranin O/fast green, or Toluidine blue staining. The bone destruction was examined via X-ray and micro-CT analysis. Immunohistochemical analysis or TRAP staining were used to investigate the role of PSTPIP2 in osteoclasts and the expression of PSTPIP2 in synovial tissue. RT-qPCR and ELISA were used to examine the expression of pro-inflammatory cytokines in synovial tissue or serum. AIA rats were found to have decreased PSTPIP2 expression and AIA-associated bone loss and inflammatory infiltration. We showed that administration of AAV-PSTPIP2 before arthritis onset significantly reduces the severity of AIA. PSTPIP2 was highly expressed in synovial cells. In addition, inflammatory responses and the number of osteoclasts were reduced with AAV-PSTPIP2 treatment. These findings demonstrate that PSTPIP2 may improve the severity of AIA by inhibiting the function of fibroblast-like synoviocytes, suppressing inflammation and reducing the number of osteoclasts.
脯氨酸-丝氨酸-苏氨酸磷酸酶相互作用蛋白 2（PSTPIP2）与炎症有关。在这项研究中，我们通过使用腺相关病毒（AAV）在大鼠中过表达 PSTPIP2 来研究 PSTPIP2 在佐剂诱导关节炎（AIA）中的作用。在第 0 天向 Lewis 大鼠注射完全弗氏佐剂（CFA）来制备 AIA 大鼠。在诱导后第 8 天，将 AAV-空或 AAV-PSTPIP2 或 PBS 关节内注射到每个膝关节中。所有动物均在佐剂注射后第 18 天处死。WB 用于检测大鼠滑膜组织中 PSTPIP2 的表达。荧光显微镜显示滑膜组织中的转导效率。通过 HE、番红 O/快绿或甲苯胺蓝染色检查关节炎关节的形态。通过 X 射线和微 CT 分析检查骨破坏。免疫组织化学分析或 TRAP 染色用于研究 PSTPIP2 在破骨细胞中的作用以及 PSTPIP2 在滑膜组织中的表达。RT-qPCR 和 ELISA 用于检测滑膜组织或血清中促炎细胞因子的表达。发现 AIA 大鼠的 PSTPIP2 表达降低，与 AIA 相关的骨丢失和炎症浸润减少。我们表明，在关节炎发作前给予 AAV-PSTPIP2 可显著减轻 AIA 的严重程度。PSTPIP2 在滑膜细胞中高表达。此外，用 AAV-PSTPIP2 治疗可减少炎症反应和破骨细胞数量。这些发现表明，PSTPIP2 可能通过抑制成纤维样滑膜细胞的功能、抑制炎症和减少破骨细胞数量来改善 AIA 的严重程度。
