TLR4/NF-κB通路参与大鼠膝骨关节炎滑膜早期病变的研究
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摘要
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 type Ⅲ procollagen(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-α and 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.
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 type Ⅲ procollagen(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-α and 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.
引文
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[8] Robinson WH,Lepus CM,Wang Q,et al. Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis[J]. Nat Rev Rheumatol,2016,12(10):580-592.
[9]陈树清,孙保国,周厚明,等.活血利水中药联合复方南星止痛膏治疗慢性膝关节滑膜炎的病例对照研究[J].中国骨伤,2012,25(4):283-286.CHEN SQ,SUN BG,ZHOU HM,et al. Combined use of Compound Nanxing Pain Paste with blood-promoting and diuretic Chinese herbal medicines in treatment of chronic knee synovitis[J]. Zhongguo Gu Shang/China J Orthop Trauma,2012,25(4):283-286. Chinese with abstract in English.
[10] de Lange-Brokaar BJ,Ioan-Facsinay A,Yusuf E,et al. Association of pain in knee osteoarthritis with distinct patterns of synovitis[J].Arthritis Rheumatol,2015,67(3):733-740.
[2] Nielsen FK,Egund N,Jorgensen A,et al. Risk factors for joint replacement in knee osteoarthritis;a 15 year follow up study[J]. BMC Musculoskelet Disord,2017,18(1):510.
[3] Zeng C,Li YS,Lei GH. Synovitis in knee osteoarthritis:a precursor or a concomitant feature[J]. Ann Rheum Dis,2015,74(10):e58.
[4] Wang H,Wang Q,Yang M,et al. Histomorphology and innate immunity during the progression of osteoarthritis:Does synovitis affect cartilage degradation[J]. J Cell Physiol,2018,233(2):1342-1358.
[5] Nair A,Kanda V,Bush Joseph C,et al. Synovial fluid from patients with early osteoarthritis modulates fibroblast like synoviocyte responses to toll like receptor 4 and toll like receptor 2 ligands via soluble CD14[J]. Arthritis Rheum,2012,64(7):2268-2277.
[6] Hulth A,Lindberg L,Telhag H. Experimental osteoarthritis in rabbits. Preliminary report[J]. Acta Orthop Scand,1970,41(5):522-530.
[7] Ghosh S,Dass JF. Study of pathway cross-talk interactions with NFkappaB leading to its activation via ubiquitination or phosphorylation:a brief review[J]. Gene,2016,584(1):97-109.
[8] Robinson WH,Lepus CM,Wang Q,et al. Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis[J]. Nat Rev Rheumatol,2016,12(10):580-592.
[9]陈树清,孙保国,周厚明,等.活血利水中药联合复方南星止痛膏治疗慢性膝关节滑膜炎的病例对照研究[J].中国骨伤,2012,25(4):283-286.CHEN SQ,SUN BG,ZHOU HM,et al. Combined use of Compound Nanxing Pain Paste with blood-promoting and diuretic Chinese herbal medicines in treatment of chronic knee synovitis[J]. Zhongguo Gu Shang/China J Orthop Trauma,2012,25(4):283-286. Chinese with abstract in English.
[10] de Lange-Brokaar BJ,Ioan-Facsinay A,Yusuf E,et al. Association of pain in knee osteoarthritis with distinct patterns of synovitis[J].Arthritis Rheumatol,2015,67(3):733-740.