Protective Effect of Yiqi Yangyin Huoxue Recipe(益气养阴活血方)Regulating Nrf2/NLRP3 Signaling Pathway on Kidney of Diabetes Nephropathy Rats
Objective To explore the renal protective mechanism of Yiqi Yangyin Huoxue Recipe(益气养阴活 血方)on dia-betes nephropathy(DN)rats by regulating nuclear factor E2 related factor 2(Nrf2)/NOD like receptor protein 3(NLRP3)signal pathway.Methods Forty male SD rats of clean grade were randomly divided into blank group,model group,valsartan group,equal dose group of Yiqi Yangyin Huoxue Recipe and high dose group after adaptive feeding for one week.Except the blank group,the rats in each group were fed with high fat and high sugar,combined with a single injection of streptozotocin(STZ)to establish the DN rat model.After successful modeling,the two groups of Chinese medicine were given Yiqi Yangyin Huoxue Recipe by gavage.The valsartan group was given valsartan by gavage,the blank group and the model group were given normal saline by gavage,and 24 hours urine,blood and kidney tissue of rats were taken out after 6 weeks.Biochemical tests were carried out to determine 24 h urine total protein(24 h-UTP),blood creatinine(SCR),blood urea nitrogen(BUN),glucose(GLU),total cholesterol(CHO)and triglyceride(TG)of rats in each group.The serum levels of interleukin 1β(IL-1 β)and interleukin 18(IL-18)were de-tected by enzyme-linked immunosorbent assay(ELISA).HE staining and light microscopy were used to observe the pathological changes of renal tissue.Protein levels of Nrf2,heme oxygenase-1(HO-1)and NLRP3 in renal tissues were detected by West-ern blot.The expressions of Nrf2,HO-1 and NLRP3 mRNA in the kidney of rats were detected by real-time fluorescence quantitative polymerase chain reaction(PCR).Results Biochemistry:Compared with those of the blank group,the levels of 24 h-UTP,Scr,BUN,GLU,CHO and TG in the model group were significantly increased(P＜0.01).Compared with those of the model group,the levels of 24 h UTP,Scr,BUN,GLU,CHO and TG in each intervention group were significantly reduced(P＜0.05 or P＜0.01).ELISA:Compared with those of the blank group,the serum levels of IL-1β and IL-18 in the model group increased significantly(P＜0.01).Compared with those of the model group,the serum levels of IL-1β and IL-18 of each in-tervention group decreased significantly(P＜0.05 or P＜0.01).Renal pathology:Compared with those of the blank,the patho-logical changes of renal tissue in the model group were serious.Compared with those of the model group,the renal pathology of each intervention group was improved.Western Blot and PCR:Compared with those of the blank group,the expressions of Nrf2 mRNA,HO-1 mRNA and Nrf2,HO-1 protein in the kidney of the model group decreased(P＜0.05 or P＜0.01),and the ex-pressions of NLRP3 mRNA and NLRP3 protein increased(P＜0.05 or P＜0.01).Compared with those of the model group,the expressions of Nrf2 mRNA,HO-1 mRNA and Nrf2,HO-1 protein in each intervention group increased(P＜0.05 or P＜0.01),and the expressions of NLRP3 mRNA and NLRP3 protein decreased(P＜0.05 or P＜0.01).Conclusion Yiqi Yangyin Huoxue Recipe may improve the above biochemical indexes and alleviate the renal pathological damage by regulating Nrf2/NL-RP3 signal pathway,reducing oxidative stress and inhibiting the inflammatory reaction of DN rats.
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