温阳通络胶囊干预骨关节炎大鼠软骨细胞的炎症反应和血管新生.

BACKGROUND: Wenyang Tongluo capsule has obvious clinical effects in the treatment of osteoarthritis, but its mechanism of action is unclear. OBJECTIVE: To investigate the therapeutic effect of Wenyang Tongluo capsule on osteoarthritis and its regulation mechanism on microRNA-140 (miR-140)/ vascular endothelial growth factor (VEGF) signaling in chondrocytes. METHODS: (1) Animal experiments: A total of 75 Sprague-Dawley rats were randomized into 5 groups (n=15 per group): normal group, model group, drug group, inhibitor group, and interference group. In the latter three groups, miR-140-antagomir NC and siRNA-VEGF NC, miR-140-antagomir and siRNA-VEGF NC, and miR-140-antagomir and siRNA-VEGF were injected into the left hindlimb joint daily, respectively. Administration in each group lasted for 2 weeks. Afterwards, the rats in the drug, inhibitor, and interference groups were intragastrically given Wenyang Tongluo capsule for 4 continuous weeks. Animal models of osteoarthritis were prepared via the intraarticular injection of papain on days 1, 4, and 7 in the model, drug, inhibitor, and interference groups. After the completion of drug administration, related measurements were performed. (2) Cell assays: ATDC5 chondrocytes were divided into five groups: normal group (cultured in the medium containing miR-140-antagomir NC and siRNA-VEGF NC), model group (cultured in the medium containing interleukin-1β, miR-140- antagomir NC and siRNA-VEGF NC, drug group (cultured in the medium containing interleukin-1β, miR-140-antagomir NC, siRNA-VEGF NC, and Wenyang Tongluo capsule), inhibitor group (cultured in the medium containing interleukin-1β, miR-140-antagomir and Wenyang Tongluo capsule), and interference group (cultured in the medium containing interleukin-1β, miR-140-antagomir, siRNA-VEGF and Wenyang Tongluo capsule). Relevant detections were then performed. RESULTS AND CONCLUSION: (1) Animal experiments: Compared with normal rats, the expression of miR-140 mRNA in cartilage tissue of osteoarthritic rats was decreased (P < 0.05), while the expression of vascular endothelial growth factor mRNA was increased (P < 0.05). Compared with the model group, interleukin-6 and tumor necrosis factor-α levels, chondrocyte apoptosis, the number of new blood vessels and protein expression of vascular endothelial growth factor in serum of the drug group, inhibitor group and interference group were significantly decreased (P < 0.05). The drug group could most significantly interfere with the malignant progression of these indicators. (2) Cell assays: Compared with normal chondrocytes, the mRNA expression of miR-140 in osteoarthritc chondrocytes was decreased (P < 0.05), while the mRNA expression of vascular endothelial growth factor was increased (P < 0.05). Compared with the model group, the proliferation activity was increased in the drug group, inhibitor group and interference group (P < 0.05), while cell apoptosis, interleukin-6 and tumor necrosis factor-α levels in the supernatant, the number of new blood vessels, and the expression of vascular endothelial growth factor gene and protein were significantly decreased (P < 0.05). The drug groups had the most obvious effects on these indicators. In conclusion, Wenyang Tongluo capsule can relieve the pathological damage of cartilage in osteoarthritic rats by significantly inhibiting inflammatory stress and angiogenesis in chondrocytes, which may be related to the regulation of the miR-140/VEGF signal axis. [ABSTRACT FROM AUTHOR]