TLR4/NF-κB通路参与大鼠膝骨关节炎滑膜早期病变的研究

Role of TLR4/NF-κB pathway for early change of synovial membrane in knee osteoarthritis rats

目的:探讨TLR4/NF-κB通路在大鼠膝骨关节炎(OA)滑膜早期病变的表达情况。方法:将8周龄体重为(200±20) g的18只雄性SD大鼠按照随机数字表法分为模型组和对照组,每组9只。模型组按照改良Hulth法构建膝OA模型,对照组不做手术。分别于术后4、 21 d提取滑膜组织和血清,采用PCR法检测CD14、TLR-4、IL-1β、TNF-α、MMP-13、ADAMTS-4表达;采用Western blot法检测NF-κB p65蛋白表达;采用Elisa法检测血清中透明质酸(HA)及Ⅲ型前胶原氨基端(PⅢNP)浓度。结果:术后4、21 d时模型组CD14、ADAMTS-4及NF-κB p65表达均较对照组升高,术后21 d时模型组TLR-4、Ⅱ-1β、TNF-α及MMP-13表达较对照组升高(P<0.01)。模型组术后4 d时血清PⅢNP和HA浓度高于对照组,术后21 d时两组比较差异无统计学意义。结论:NF-κB通路可通过CD14/TLR-4早期激活进而触发滑膜分泌炎性因子IL-1β、TNF-a、MMP-13、ADAMTS-4,PⅢNP和HA增加介导膝OA发生。

Objective:To study role of TLR4/NF-κB pathway for early change of synovial membrane in knee osteoarthritis rats.Methods:Eighteen male SD rats weighted(200±20)g were randomly divided into 2 groups,namely control and model group,and 9 in each group.Knee OA model group was established by using modified Hulth method in model group.Control group was not treated.Synovial tissue and serum was extracted at 4 and 21 d after operation.Expression of CD14,TLR4,IL-1β,TNF-α,ADAMTS-4,MMP-13 were detected by real-time PCR respectively.NF-κB p65 protein was detected by Western-blot;serum concentrations of haluronic acid(HA),N-propeptide of typeⅢprocollagen(PⅢNP)was detected by Elisa.Results:Expression of CD14,ADAMTS-4,and NF-κB p65 in model group were higher than that of control group at 4 and 21 days after operation,while expression of TLR4,IL-1β,TNF-αand MMP-13 were higher than that of control group at 21 days after operation(P<0.01).Concentration of PIIINP and HA in model group were higher than that of control group at 4 days after operation,while there was no significant difference at 21 days after operation.Conclusion:NF-κB pathway could mediate occurrence of KOA by early activating and triggeringg synovial increasingly secreting inflammatory secretion CD14,TLR4,IL-1β,TNF-α,ADAMTS-4,MMP-13,PⅢNP and HA.