Study On Inflammatory Factor Of Uric Acid Nephropathy Rats By Decoction Of Clearing Damp Turbidity And Stasis Based On NLRP3 Inflammasome

Objective:To observe the effects of decoction of clearing damp turbidity and stasis on serum uric acid(SUA),serum creatinine(Scr),urea nitrogen(BUN)and renal histopathological changes in rats with uric acid nephropathy(UAN).To determine the content of IL-1beta,the expression of NLRP3 protein and the expression of NLRP3,IL-1beta and IL-18 mRNA,and to explore whether the recipe can affect the inflammatory factor IL-1beta by down-regulating the inflammatory body of NLRP3.The expression of IL-18 and IL-18 can intervene uric acid nephropathy.Methods: Sixty Wistar rats were randomly divided into six groups(blank group,model group,allopurinol group,high-dose group,medium-dose group and low-dose group)after one week of adaptive feeding,and were fed with common diet and drinkable distilled water freely.The UAN model was established on the same day as the drug administration.Adenine(100mg/kg)+ethambutol(250mg/kg)suspension was administered orally in the morning to establish the model.In the afternoon,the blank group and the model group were administered with normal saline of equal volume.The remaining groups were administered with different concentrations of prescriptions(10.53g/kg,21.07g/kg,42.14g/kg)and allopurinol suspension(27mg/kg)respectively.For 21 consecutive days.The weight of rats was weighed after 2 hours of intragastric administration on the 21 st day of model establishment and anesthetized according to the weight.The pathological changes of rat kidney were observed by SUA,Scr,BUN,HE staining.The serum IL-1beta content was detected by ELISA,the protein production of NLRP3 was detected by immunohistochemistry,and the expression of NLRP3,IL-1beta and IL-18 in rat kidney was detected by Q-PCR.Result:1.The results of blood uric acid and renal function test showed that the levels of SUA,Scr and BUN in model group were significantly higher than those in blank group(P<0.01).Compared with the model group,SUA levels in the high,middle and allopurinol groups were significantly lower(P<0.05),while SUA levels in the low dose group were slightly lower(P>0.05),but the difference was not statistically significant(P>0.05).The levels of Scr and BUN in each dose group and allopurinol group were significantly decreased(P<0.05).2.Serum IL-1beta content: Compared with the model group,the expression of IL-1beta in the high-dose,middle-dose and allopurinol groups decreased significantly(P < 0.01),and the content of IL-1beta in the middle-dose group was the closest to that in the blank group(P < 0.01).3.The results of HE staining under light microscopy showed that the pathological changes of kidney tissue in model group were more obvious than that in blank group.The lumen was dilated,the epithelial cells of renal tubules fell off,necrosed,disintegrated and fell into the lumen.In renal tubules and interstitium,deposition of brown urate crystals and local infiltration of inflammatory cells were observed.Urate crystallization in kidneys of rats in high,medium and low dose groups and allopurinol groups were reduced in varying degrees.Inflammatory cell infiltration in kidney tissue was alleviated in varying degrees,and renal tubular dilatation was improved.Among them,the kidney tissue structure of the middle dose group was relatively intact,and the degree of cell damage was mild,which was closest to the pathological section of the normal group.4.Immunohistochemical results showed that there was no significant expression of NLRP3 protein in blank group,and the positive expression of NLRP3 in model group was significantly higher than that in blank group(P<0.01);the positive expression of NLPR3 protein was brown lump,mainly distributed in renal tubules and interstitium;the expression of NLRP3 protein in each drug intervention group was lower than that in model group(P<0.01);the effect of reducing the expression of NLRP3 protein in medium dose group of Chinese medicine was better than that in other groups(P<0.01).The purinol group(P<0.05),and the middle dose group of traditional Chinese medicine had the best effect on down-regulation of NLRP3 protein expression in each drug intervention group(P<0.01).5.The results of Q-PCR showed that the expression levels of IL-1beta,IL-18 and NLRP3 in kidney tissue of rats in each group were higher than those in blank group.Compared with model group,the expression levels of IL-1beta,IL-18 and NLRP3 in kidney tissue of rats in each treatment group were significantly lower(P<0.01),and the level of IL-1beta,IL-18 and NLRP3 in middle dose group was lower than that in other drug intervention group.Conclusion:1.Decoction of clearing damp turbidity and stasis can effectively improve the general state of UAN rats,reduce the level of SUA,Scr and BUN in UAN rats,reduce the deposition of uric acid in kidney,reduce the degree of degeneration,inflammatory infiltration and necrosis of kidney cells,and have a certain protective effect on kidney.2.Decoction of clearing damp turbidity and stasis significantly reduced the content of IL-1beta,the expression of NLRP3 protein and the expression of NLRP3,IL-1beta and IL-18 mRNA,thereby alleviating the degree of UAN lesion.3.By inhibiting the activation of NLRP3 inflammatory body,inhibiting the secretion and release of inflammatory factors IL-1beta and IL-18,the decoction of clearing damp turbidity and stasis can interfere with UAN,which may be one of the decoction of clearing damp turbidity and stasis in treating UAN.