通络止痛方对人膝骨关节炎滑膜炎性细胞TLR的影响

Effect of Tongluo Zhitong Formulation on Toll like receptor of synovitis cells of human knee osteoarthritis

关键词:  Toll样受体  通络止痛方  膝骨关节炎  滑膜炎症

Key words:  Toll like receptor  Tongluo Zhitong Formulation  knee osteoarthritis  synovial inflammation

摘要:

[目的] 观察通络止痛方对人膝骨关节炎滑膜炎性细胞Toll样受体（TLR）的影响。[方法] 取院内行全膝关节置换术患者术中废弃滑膜组织，进行原代培养，取3~4代成纤维细胞进行不同浓度浓度通络止痛方干预，具体分为：高浓度组（2.5 mg/mL）、中浓度组（0.25 mg/mL）、低浓度组（0.025 mg/mL）、对照组（0 mg/mL）。应用real-time PCR检测细胞内TLR4 mRNA表达情况，应用流式细胞术（FCM）检测细胞中TLR2、TLR4表达情况。[结果] 与对照组比较，高、中浓度组通络止痛方人膝骨关节炎滑膜炎症细胞TLR4 mRNA的表达水平显著降低（P<0.05）。低浓度组通络止痛方对滑膜炎症细胞TLR4 mRNA抑制作用与对照组相近，差异无统计学意义。中、高浓度组抑制作用与浓度呈正比。高、中、低浓度通络止痛方组TLR4表达率与对照组比较都有明显的降低（P<0.05），且随着浓度升高，TLR4的表达率逐渐降低；高、中、低浓度通络止痛方组TLR2表达率与对照组比较差异无统计学意义（P>0.05）。[结论] 高、中、低浓度通络止痛方均可以显著抑制Toll样受体的表达，提示通络止痛方可能通过Toll样受体通路影响或抑制人膝骨关节炎滑膜炎症反应。

Abstract:

[Objective] To observe the effect of Tongluozhitong Formulation on the Toll like receptor of synovitis cells of human knee osteoarthritis.[Methods] Our hospital waste in synovial tissue of patients with total knee replacement surgery,were cultured from 3~4 fibroblasts by different concentration of Tongluozhitong Formulation,specifically divided into:high concentration group (2.5 mg/mL),medium concentration group (0.25 mg/mL);low concentration group (0.025 mg/mL);control group (0 mg/mL). The expression of TLR4 mRNA in cells was detected by al-time PCR,and the expression of TLR2 and TLR4 in cells was detected by flow cytometry (FCM).[Results] Compared with the control group,the expression level of TLR4 mRNA in the knee osteoarthritis synovitis cells of the high and middle concentration group Tongluozhitong Formulation was significantly lower than that of the control group (P<0.05). The inhibitory effect of Tongluozhitong Formulation in low concentration group on TLR4 mRNA of synovitis was similar to that of the control group,and the difference was not statistically significant. The inhibitory effect of medium and high concentration groups was proportional to the concentration. High,medium and low concentration of Tongluozhitong Formulation group the expression rate of TLR4 compared with the control group were significantly decreased (P<0.05),and with the concentration of TLR4,the expression rate decreased;high,medium and low concentration of Tongluozhitong Formulation group the expression rate of TLR2 compared with the control group,the difference was not statistically significant (P>0.05).[Conclusion] High,middle and low concentrations Tongluozhitong Formulation can significantly inhibit the expression of Toll like receptor,suggesting that Tongluozhitong Formulation can affect or inhibit synovial inflammation of human knee osteoarthritis through Toll like receptor (TLR) pathway.