■在这项研究中,我们研究了维生素D(VITD)在前交叉韧带横断联合内侧半月板切除术(ACLTMMx)诱导的OAWistar大鼠中的治疗潜力。在ACLT+MMx诱导的OA大鼠中,疼痛严重程度,软骨破坏,炎性细胞因子,和MMPs都进行了测量。
■ACLT+MMx方法用于诱导OA,并进行了疼痛行为研究,如负重试验和缩爪试验,同时还测量了膝盖宽度和体重。此外,苏木精和伊红(H&E)染色用于确定膝关节组织病理学研究,以及OARSI评分,软骨厚度,软骨宽度,和软骨退化评分。采用酶联免疫吸附试验(ELISA)检测血清VITD水平,II型胶原的C端肽(CTX-II),和促炎细胞因子肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β),白细胞介素-6(IL-6),和抗炎细胞因子白细胞介素-10(IL-10),和MMP(MMP-3、MMP-9和MMP-13)。最后,逆转录聚合酶链反应(RT-PCR)测试用于确定MMPs的水平,核因子-κB(NF-κB),TNF-α,IL-1β刺激的软骨细胞中的IL-6和IL-10。
■口服VITD补充剂可显着减轻OA疼痛,炎症,软骨破坏,和MMPs水平。此外,血清VITD水平升高而CTX-II水平降低,表明VITD能有效降低软骨降解。此外,补充VITD可降低促炎TNF-α的表达,IL-1β,和IL-6细胞因子,同时增加抗炎IL-10的表达。ACLT+MMx手术后MMPs的升高导致关节软骨的破坏,通过补充VITD减少。最后,VITD补充显著降低血清MMPs水平,IL-1β,TNF-α,和IL-6,同时增加IL-10水平。然后,使用体外细胞毒性(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物)MTT测定法,检测IL-1β刺激后大鼠软骨细胞中VITD的细胞毒性谱,在VITD0-500IU的剂量范围内没有毒性。最后,IL-1β刺激的大鼠软骨细胞的RT-PCR研究显示,VITD(50、100和500IU)显着降低MMPs的mRNA水平,NF-κB,TNF-α,和IL-6,同时增加IL-10水平,表明VITD以剂量依赖的方式减少了软骨细胞的破坏并克服了恶劣的条件。
■总的来说,体内和体外研究结果表明,VITD可有效减轻OA疼痛,炎症,和软骨细胞的破坏通过降低MMP水平特异性。
UNASSIGNED: In this study, we investigated the therapeutic potential of vitamin D (VITD) in OA Wistar rats induced by anterior cruciate ligament transection combined with medial meniscectomy (ACLT + MMx). In ACLT + MMx-induced OA rats, pain severity, cartilage destruction, inflammatory cytokines, and MMPs were all measured.
UNASSIGNED: ACLT + MMx methods were used to induce OA, and pain behavioral studies such as the weight bearing test and paw withdrawal test were performed while the knee width and body weights were also measured. Furthermore, Hematoxylin and Eosin (H&E) staining was used to determine knee histopathological studies, as well as OARSI scoring, cartilage thickness, cartilage width, and cartilage degradation scores. The enzyme-linked immunosorbent assay (ELISA) studies were used to check the serum levels of VITD, C-telopeptide of Type II collagen (CTX-II), and pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and anti-inflammatory cytokines interleukin-10 (IL-10), and MMPs (MMP-3, MMP-9, and MMP-13). Finally, the reverse transcription polymerase chain reaction (RT-PCR) test was used to determine the levels of MMPs, nuclear factor-kappa B (NF-κB), TNF-α, IL-6, and IL-10 in IL-1β stimulated chondrocytes.
UNASSIGNED: The oral VITD supplement significantly reduced OA pain, inflammation, cartilage destruction, and MMPs levels. Furthermore, serum VITD levels increased while CTX-II levels decreased, indicating that VITD reduced cartilage degradation effectively. Moreover, VITD supplementation reduced the expression of pro-inflammatory TNF-α, IL-1β, and IL-6 cytokines while increasing the expression of anti-inflammatory IL-10. The elevation of MMPs after ACLT + MMx surgery contributed to articular cartilage destruction, which was reduced by VITD supplementation. Finally, VITD supplementation significantly reduces serum levels of MMPs, IL-1β, TNF-α, and IL-6 while increasing IL-10 levels. Then, using the in-vitro cytotoxicity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) MTT assay, examine the cytotoxicity profile of VITD in rat chondrocytes after stimulated with IL-1β, which shows no toxicity in the dose range of VITD 0-500 IU. Finally, RT-PCR studies in IL-1β stimulated rat chondrocytes revealed that VITD (50, 100, and 500 IU) significantly reduced the mRNA levels of MMPs, NF-κB, TNF-α, and IL-6, while increasing IL-10 levels, indicating that VITD reduced chondrocyte destruction and overcame harsh conditions in a dose-dependent manner.
UNASSIGNED: Overall, the in vivo and in vitro findings show that VITD effectively reduces OA pain, inflammation, and chondrocyte destruction by lowering MMPs levels specifically.