Regulation Of Modified Simiao Decoction On TLR4-NF-κB And NLRP3 Inflammasome Signaling Pathways In Rat With Acute Gouty Arthritis

Objective:Establish acute gouty arthritis model in rats,and rats in different treatment groups were given by corresponding medicine.By investigating the inflammatory cells,proteins of TLR4-NF-κB and NLRP3 inflammasome signaling pathways and the expression of inflammatory cytokines,to explore the effect of modified simiao decoction treatment of acute gouty arthritis and the mechanisms.Methods:1.The model of acute gouty arthritis group(model group) and Blank control group(BC group) in rats were established by injecting 50 ul monosodium urate suspension liquid crystals or PBS into the right ankles of rats.After the model were established about 24 h,observed the joint swell degree,collected joint leachate to measure the the expression of IL-1β and TNFα,collected soft tissues surrounding the joint to observe inflammatory cells with histopathologic examination between the two groups of rats and detect the mRNA and protein expression level of each part in TLR4-NF-κB and NLRP3 inflammasome signaling pathways with real-time quantitative PCR and Western Blot.2.After established the modle by injecting monosodium urate suspension liquid crystals into the right ankles of rats,acute gouty arthritis rats were randomly divided into model group, colchicin group(PC), low dosage group of modified simiao decoction(MSD1),medium dosage group of modified simiao decoction(MSD2).Rats in different groups were given by corresponding medicine from gastric tube for three days. Meanwhile blank controlled group(BC) was established.We got joint leachate to measure the the expression of IL-1β、TNFα and IL-6 with enzyme-linked immunosorbent assay method,soft tissues surrounding the joint were also collected to observe inflammatory cells with histopathologic examination,what’s more the mRNA and protein expression level of each part in the TLR4-NF-κB and NLRP3 inflammasome signaling pathways were detected with real-time quantitative PCR and Western Blot in different groups of rats.Results:1.After the model were established about 24 h,the joints of model group were markedly swollen than BC group;Histopathological results show that BC group have no obvious inflammatory cell infiltration,while the model group diffuse a great deal of inflammatory cells infiltration,mainly as neutrophil、macrophages and lymphocytes.Inflammatory factor index:the IL-1β and TNF-α value of model group, were significantly higher than BC group,(P<0.05,P<0.01).When compared with BC group,the mRNA expression level of TLR4、NLRP3、ASC、Caspase-1 were significantly higher(P<0.01) and so did the protein expression level of TLR4、NF-κB、ASC、Caspase-1(P<0.01).2.Inflammatory factor index results after treatment:When compared with modle group,the IL-1β、TNF-α、IL-6 value of MSD1 group、MSD2 group and PC group,were significantly lower,more have a statistically significant difference between;When it comes to MSD1 group and MSD2 group,the IL-1β and IL-6 value of PC group had no significant difference,but the TNF-ɑ value were significantly lower(P<0.05); As for MSD1 group,the IL-1β 、 TNF-α and IL-6 value of the MSD2 group were no statistical difference.3.Histopathological results after treatment:BC group and model group displayed the same situation as part 1.When compared with modle group,there were less inflammatory cells in MSD1 group and MSD2 group;what’s more the inflammatory cells in PC group were significantly less than those in MSD1 group and MSD2 group.4. signaling pathways mRNA index results after treatment:When compared with modle group,ASC、caspase-1 mRNA expression level of MSD1 and MSD2 group were significantly lower,while the TLR4 and NLRP3 mRNA expression level were no statistical difference,and TLR4 、 NLRP3 、 ASC 、 Caspase-1 mRNA expression level of PC group were significantly lower.When it comes to TLR4、NLRP3、ASC、Caspase-1 mRNA expression level of PC group were significantly lower than MSD1 group and MSD2 group,with a statistically significant difference.As to ASC and Caspase-1 mRNA expression level of MSD2 group,they were significantly lower than MSD1 group,while there were no statistical difference of the TLR4 and NLRP3 mRNA expression level between them.5.signaling pathways protein index results after treatment:When compared with modle group,and NF-κB、caspase-1 protein expression level of MSD1 group were significantly lower,at the same time,NF-κB、ASC、caspase-1 protein expression level of MSD2 group were also significantly lower,and TLR4 、 NF-κB 、 ASC 、 Caspase-1 protein expression level of PC group were significantly lower;When it comes to PC group,NF-κB、Caspase-1 protein expression level of MSD1 group and ASC protein expression level of MSD2 group,were no statistical difference.while TLR4 protein of PC group was significantly lower than MSD1 and MSD2 group,and ASC protein expression level of PC group were significantly lower than MSD1 group(P<0.05),with a statistically significant difference.When compared with MSD1 group,only ASC protein expression level of MSD2 group was significantly lower(P<0.05),but there were no statistical difference among the expression level of TLR4、NF-κB、Caspase-1 protein.Conclusions:1.The acute gouty arthritis of wistar rats can be successfully induced by urate sodium crystal,the mechanism of it is:Through the activation of TLR4-NF-κB and NLRP3 inflammasome signaling pathways by MSU,and active Interleukin1 β(IL-1β) is produced and released,then induce acute inflammation.What’s more it can imitate acute gouty arthritis of human welly.2.Modified simiao decoction can suppress the aggregation of inflammatory cells and the expression level of inflammatory factors such as IL-1β、TNF-ɑ and IL-6,to relieve the acute inflammatory reaction of gouty arthritis.3.Modified simiao decoction can regulate the mRNA and protein expression level of TLR4-NF-κB and NLRP3 inflammasome signaling pathways with multitarget to cure acute gouty arthritis.