Inflammatory pathways and Toll-like receptors related protein expression on acute gouty arthritis model rats / 中华风湿病学杂志

ABSTRACT

Objective:Sodium urate was used to induce acute gouty arthritis rat model, and to observe the inflammatory response of rats and the intervention effect of diclofenac sodium on the expression of Toll-like receptor-related (TLR) protein of ankle joint.Methods:Thirty males specific pathogen free (SPF) grade Wistar rats were used to develop the models. Random number table method was used to divide the rats into normal saline control group, model group, and drug group (diclofenac sodium t 1.35 mg/g body weight), 10 rats in each group. After fully grinding the sodium urate crystals, an appropriate amount of saline and Tween-80 (9∶1) was added to make a suspension, and the sodium urate crystals (25 mg/ml) were injected to the right posterior ankle of the rats in the model and drug groups. The solution was 0.2 ml, and rats in the sham group were injected with 0.2 ml of normal saline at the same location. After the model was established, drug and equal volume of purified water were administrated intragastrically once a day for 7 days. The toe volume device was used to measure the joint swelling of the rat (at 4 h, 8 h, 24 h, 48 h, 72 h) , and blood was taken from the abdominal aorta after anesthesia to determine the rat kidney function, interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) content, the rat ankle joint TLR4, myeloid differentiation factor (MyD88), NF-κBp65 protein expression were determined using Western blot and immunohistochemical methods. Multiple comparisons were carried out using single factor analysis of variance (ANOVA), comparing the two groups by using LSD- t, the comparison of different times using repetitive measure analysis of variance (repeated measures). Results:After the models were established, the rat's right ankle joint showed various degrees of redness, slow walking, and unresponsiveness. Compared with the normal saline control group, under the light microscope, the ankle synovial cells of the model group proliferated, with localized degeneration and necrosis, and many inflammatory cell infiltration. The rat serum inflammatory factors IL-1β, IL-6, TNF-α in the diclofenac sodium group [(24.6±3.3) pg/ml, (151±21) pg/ml, (61±16) pg/ml] were significantly reduce compared with model group [(28.4±4.3) pg/ml, (173±26) pg/ml, (81±5) pg/ml] ( t=2.296, P<0.01; t=2.909, P<0.01; t=2.352, P<0.01). Compared with normal saline group, variance analysis showed that the NF-κBp65, MyD88, TLR4 protein expression of ankle joint detected by Western bolt method and immunohistochemistry method was significantly increased in the model group. Compared with the model group, diclofenac sodium the ankle tissue protein expression of NF-κBp65, MyD88, and TLR4 was significantly inhibited. There were statistical significances in three groups ( P<0.05 or P<0.001). Conclusion:The level of inflammatory factors in acute gout arthritis rats model induced by sodium urate crystals is increased, and the expression of TLR4/MyD88/NF-КBp65 proteins in ankle joint tissue is increased, which affects the TLR signaling pathway. Diclofenic has inhibitory and relieving effects.