塞来昔布对骨关节炎大鼠模型抗炎镇痛作用研究

Anti-inflammatory and analgesic effects of celecoxib for osteoarthritis rat model

目的研究塞来昔布对骨关节炎大鼠模型的抗炎镇痛作用,分析其作用机制。方法将50只SD大鼠通过切断前交叉韧带及内侧副韧带构建骨关节炎大鼠模型并随机分为模型组及实验组,各25只;另随机选取25只SD大鼠作为假手术组,仅打开关节腔,但不切断交叉韧带,术后实验组大鼠灌胃给予24 mg·kg^-1塞来昔布,模型组及假手术组则灌胃等量的生理盐水,每天1次,各组均连续干预4周。以酶联免疫吸附(ELISA)法检测关节液中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)及前列腺素E2(PGE2)含量,以硝酸还原酶法检测一氧化氮(NO)含量;以苏木精-伊红(HE)染色及甲苯胺蓝染色观察大鼠关节软骨病理学形态变化;以蛋白质印迹(WB)法检测关节软骨组织Toll样受体4(TLR4)、核转录因子-κB(NF-κB)蛋白表达。结果假手术组、模型组及实验组大鼠关节液IL-1β含量分别为(30.04±4.12),(82.46±6.87),(51.22±5.33)ng·L^-1;TNF-α含量分别为(24.11±3.95),(50.03±5.02),(37.45±4.23)ng·L^-1;PGE2含量分别为(1.72±0.61),(3.16±0.74),(2.51±0.70)μmol·L^-1;NO含量分别为(33.41±4.33),(152.09±10.56),(71.46±5.94)μmol·L^-1。与假手术组比,模型组关节液IL-1β、TNF-α、PGE2、NO含量及关节软骨组织TLR4、NF-κB蛋白相对表达量均显著升高,而实验组较模型组显著降低(均P<0.05)。结论塞来昔布对骨关节炎大鼠的抗炎镇痛作用显著,其作用机制可能与有效抑制TLR4、NF-κB蛋白表达,干扰TLR4/NF-κB通路信号转导相关。

Objective To explore the anti-inflammatory and analgesic effects of celecoxib for osteoarthritis rat model and analyze the action mechanism.Methods Fifty SD rats were built osteoarthritis rat model by cutting off the anterior cruciate ligament and medial ligaments and were divided randomly into model group and test group,25 cases in each group;another 25 SD rats were selected as sham-operation group,just opening the articular cavity,but not to cut off the cruciate ligament.After operation,rats in test group were gavage with 24 mg·kg^-1 celecoxib,rats in sham-operation group and model group were gavage with the same amount of normal saline,1 time·d^-1,each group was intervened for 4 weeks.The levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and prostaglandin E2(PGE2)in joint fluid were detected by enzyme-linked immunosorbent assay(ELISA),and the nitric oxide(NO)contents in joint fluid were detected by nitrate reductive enzymatic;the articular cartilage pathological morphological changes of rats were observed by hematoxylin and eosin(HE)staining and toluidine blue staining;the Toll-like receptor 4(TLR4)and nuclear transcription factor-κB(NF-κB)expression in articular cartilage were detected by Western Blot(WB).Results The joint fluid IL-1βcontents in sham-operation group,model group and test group were(30.04±4.12),(82.46±6.87),(51.22±5.33)ng·L^-1;TNF-αcontents were(24.11±3.95),(50.03±5.02),(37.45±4.23)ng·L^-1;PGE2 contents were(1.72±0.61),(3.16±0.74),(2.51±0.70)μmol·L^-1;NO contents were(33.41±4.33),(152.09±10.56),(71.46±5.94)μmol·L^-1.Compared with sham-operation group,the IL-1β,TNF-α,PGE2,NO contents in joint fluid and the TLR4,NF-κB relative expression in articular cartilage increased significantly,while test group was lower than model group(all P<0.05).Conclusion The anti-inflammatory and analgesic effects of celecoxib for osteoarthritis rat model is remarkable,and the action mechanism may be related to down-regulation of TLR4,NF-κB protein expression,interfere the TLR4/NF-κB pathway signal transduction.