Experimental Study On Treatment Of Acute Gouty Arthritis By Plantain Based On TLRs/NF-κB Signal Transduction Pathway

Objective:The rat model of acute gouty arthritis(AGA)was established by injection of sodium urate(MSU)into the joint cavity.The effect of Plantago on the expression of protein related to TLRs/NF-κB signaling pathway in AGA rats and the mechanism of action were investigated,and the relationship between GA inflammation and TLRs/NF-κB signaling pathway was verified.Methods:Sixty male SD rats were randomly divided into 6 groups,namely control group(CG),model group(MG),colchicine group,psyllium low-dose group,Psyllium medium-dose group and Psyllium high-dose group,with 10 rats in each group.Colchicine group was gavage with 0.3mg/kg colchicine solution 3 ml;psyllium low,medium and high groups were gavage with psyllium 100 mg/kg,300 mg/kg and 500mg/kg,respectively;CG and MG groups were gavage with 3 ml normal saline.On day6 after gavage,50 rats in MG group,colchicine group,psyllium low-dose group,Psyllium medium-dose group and Psyllium high-dose group were injected with MSU as GA animal model,while CG was injected with normal saline into joint cavity.The degree of ankle joint swelling was measured before the experiment.After the successful construction of GA model,gait scores and ankle joint swelling were measured at 6 h,12 h,24 h and 48 h,respectively.After the experiment,6 ml of abdominal aorta blood was taken under anesthesia to determine the serum uric acid level,and serum IL-1β,IL-6and TNF-α levels were detected by ELISA.The histopathological observation of the rat ankle joint section was made.Western blot was used to detect the expression of TLR2,TLR4,NF-κB and IL-1β in synovium.Result:1.Gait score of rats.There was no difference in the gait score of the six groups before modeling(P >0.05).After modeling,compared with blank group,gait scores of model group,plantago low-dose group and medium-dose group were significantly different at 6 h,12 h,24 h and 48 h(P < 0.05).Gait scores of colchicine group and plantain high-dose group were different at 6 h and 12 h(P < 0.05),but there was no statistical significance at 24 h and48 h(P > 0.05).Compared with model group,there were significant differences in gait scores of colchicine group and plantain group at 6 h,12 h,24 h and 48 h(P < 0.05).Compared with colchicine group,the gait scores of plantain low-dose and medium-dose groups were different at 6 h,12 h,24 h and 48 h(P < 0.05).There was no significant difference in gait scores in high-dose plantain group at 6 h,12 h,24 h and 48 h(P >0.05).2.Changes of periankle diameter in rats.There was no difference in periankle diameter before modeling among the six groups(P > 0.05).After modeling,compared with blank group,there were statistically significant differences in ankle circumference diameter between model group,plantago low-dose group and medium-dose group at 6 h,12 h,24 h and 48 h(P < 0.05).There was significant difference in ankle circumference diameter between colchicine group and plantain high-dose group at 6 h and 12 h(P < 0.05),but no significant difference in ankle circumference at 24 h and 48 h(P > 0.05).Compared with model group,colchicine group and plantain group had statistical significance in ankle circumference diameter at 6 h,12 h,24 h and 48 h(P < 0.05).Compared with colchicine group,the ankle circumference of plantain low-dose and medium-dose groups was different at 6 h,12 h,24 h and 48 h(P < 0.05).There was no statistical significance in ankle circumference diameter at 6 h,12 h,24 h and 48 h in high-dose plantago group(P >0.05).3.Analysis of serum uric acid level in ratsCompared with blank group,there were differences among model group,Psyllium low-dose group and medium dose group(P < 0.05).There was no significant difference in serum uric acid between colchicine group and psyllium high-dose group(P > 0.05).Compared with model group,there was significant difference in serum uric acid between colchicine group and psyllium dosage group(P > 0.05).Compared with colchicine group,serum uric acid level in psyllium low-dose and medium-dose groups was different(P < 0.05).There was no statistical significance in serum uric acid level of Psyllium high-dose group(P > 0.05).4.HE staining results.Under light microscope,the synovial cells in blank group were arranged neatly.The synovium in the model group was hyperplasia and contained a large number of inflammatory cells.A few synovial hyperplasia and inflammatory cells were observed in colchicine group.The inflammatory cells decreased with the increase of plantain concentration.5.The levels of IL-1β,IL-6 and TNF-α in serum were determined by ELISA.Compared with blank group,there were differences among model group,Psyllium low-dose group and medium dose group(P < 0.05).There was no statistical difference in serum concentrations of TNF-alpha,IL-6 and IL-1β between colchicine group and plantain high-dose group(P > 0.05).Compared with model group: Serum concentrations of TNF-alpha,IL-6 and IL-1β in colchicine group and Psyllium dose group had statistical significance(P < 0.05).Compared with colchicine group,serum concentrations of TNF-alpha,IL-6 and IL-1β were different in the plantain low-dose and medium-dose groups(P < 0.05).Serum concentrations of TNF-alpha,IL-6 and IL-1β in high-dose plantago group did not have statistical significance(P > 0.05).6.Comparison of TLR2,TLR4,NF-κB and IL-1β in synovium by Western Blot analysisCompared with blank group,there were differences among model group,Psyllium low-dose group and medium dose group(P < 0.05).There were no significant differences in the expressions of TLR2,TLR4,NF-κB and IL-1β in synovium between colchicine group and plantago high-dose group(P > 0.05).Compared with model group,the protein expressions of TLR2,TLR4,NF-κB and IL-1β in synovium tissue of colchicine group and plantago dose group were statistically significant(P < 0.05).Compared with colchicine group,the protein expressions of TLR2,TLR4,NF-κB and IL-1β in synovium of plantago L.were different between low-dose and medium-dose groups(P < 0.05).The protein expressions of TLR2,TLR4,NF-κB and IL-1β in the synovium of plantago high-dose group were not statistically significant(P > 0.05).Conclusion:Plantago can improve the pathological changes of synovium of ankle in GA model rats,inhibit the expression of TLR2,TLR4,NF-κB,IL-1β,and effectively alleviate the progression of GA model rats.The mechanism may be to inhibit the increase of IL-1β,IL-6 and TNF-α through the TLRs/NF-κB signaling pathway,thus inhibiting the increase of inflammatory factors.