荣筋拈痛方含药血清对骨关节炎大鼠SDF-1/CXCR4信号通路的影响

Effects of Rongjin Niantong Fang on SDF-1/CXCR4 Signal Pathway in Rats with Osteoarthritis

目的观察荣筋拈痛方对骨关节炎(OA)大鼠SDF-1/CXCR4信号通路相关调节因子的影响,探讨荣筋拈痛方治疗OA的作用机制。方法采用4%木瓜蛋白酶注射关节腔复制大鼠OA模型,Ⅱ型胶原酶消化法获取正常滑膜细胞与OA滑膜细胞,正常滑膜细胞即空白组,OA滑膜细胞分为模型组和治疗组。空白组和模型组细胞用10%空白血清培养,治疗组用10%荣筋拈痛方含药血清培养;各组干预24 h后,采用ELISA法检测滑膜细胞上清液中SDF-1含量,Real-time PCR和Western blot法检测滑膜细胞SDF-1 mRNA和蛋白表达。收集的滑膜细胞上清液干预退变软骨细胞24 h后,Real-time PCR和Western blot法检测退变软骨细胞中CXCR4、MMP-3、MMP-9和MMP-13 mRNA和蛋白表达。结果与空白组比较,模型组大鼠滑膜细胞上清液中SDF-1含量明显增加(P<0.05),大鼠滑膜细胞SDF-1 mRNA和蛋白表达明显升高(P<0.05),退变软骨细胞CXCR4、MMP-3、MMP-9和MMP-13 mRNA和蛋白表达明显升高(P<0.05);与模型组比较,治疗组大鼠滑膜细胞上清液中SDF-1含量明显减少(P<0.05),大鼠滑膜细胞SDF-1 mRNA和蛋白表达明显降低(P<0.05),退变软骨细胞CXCR4、MMP-3、MMP-9和MMP-13 mRNA和蛋白表达明显降低(P<0.05)。结论荣筋拈痛方可通过调控SDF-1/CXCR4信号通路,抑制OA炎症反应,起到治疗OA作用。

Objective: To observe the effect of Rongjin Niantong Fang(RJNTF) on the related regulatory factors in stromal cellderived faceor-1(SDF-1)/chemokine C-X-C motif receptor 4(CXCR4) signaling pathway in rats with osteoarthritis, so as to explore its mechanism on osteoarthritis treatment. Methods: Rat models with knee osteoarthritis were established by injecting 4% papain into the knee joint cavity. The normal synoviocytes(blank group) and the osteoarthritis synoviocytes were obtained by Type Ⅱ collagenase digestion method. The osteoarthritis synoviocytes are divided into model group and treatment group. The blank group and the model group cells were cultured with 10% blank serum, and the treatment group was cultured with 10% RJNTF-contained serum. The content of SDF-1 in supernatant of synovial cells was detected by ELISA, and the m RNA and protein expression of SDF-1 was detected by Real-time PCR and Western blot respectively after treated 24 hours. After the degenerative chondrocytes were treated with the supernatant of synoviocytes for 24 h, the m RNA and protein expression of CXCR4、MMP-3、MMP-9、MMP-13 was detected by Realtime PCR and Western blot respectively. Results: Compared with the blank group, the content of SDF-1 in supernatant of synovial cells in the model group increased significantly(P<0.05), the expression of SDF-1 m RNA and protein in synovial cells increased significantly(P<0.05), and the expression of CXCR4, MMP-3, MMP-9 and MMP-13 m RNA and protein in degenerative chondrocytes increased significantly(P<0.05). Compared with the model group, the content of SDF-1 in the supernatant of synovial cells in the treatment group decreased significantly(P<0.05), the expression of SDF-1 m RNA and protein in synovial cells decreased significantly(P< 0.05), and the expression of CXCR4, MMP-3, MMP-9 and MMP-13 m RNA and protein in degenerative chondrocytes decreased significantly(P<0.05). Conclusion: RJNTF could inhibit the inflammatory reaction in osteoarthritis by regulating SDF-1/CXCR4 signaling pathway, which may explain its clinical efficacy in osteoarthritis treatment.