补肾活血泄浊方调节Keap1/Nrf2/HO-1介导的铁死亡改善慢性肾衰竭大鼠纤维化的机制研究OA

Objective To evaluate the effects of Bushen Huoxue Xiezhuo Formula(补肾活血泄浊方,BHXF)in a rat model of chronic renal failure(CRF)and investigate its potential anti-renal fibrosis mechanism related to the ferroptosis pathway mediated by Keap1/Nrf2/HO-1.Methods Ninety male SD rats were randomly divided into six groups(n=15 per group):the sham-operated group,model group,low/medium/high-dose BHXF group(5.31 g·kg-1·d-1,10.62 g·kg-1·d-1,21.24 g·kg-1·d-1),and the benazepril group(10 mg·kg-1·d-1).Rats received the corresponding doses of BHXF or benazepril by gavage once daily for 8 weeks;the sham-operated and model groups received an equivalent volume of physiological saline.After the experiment,kidney tissues were collected for histological exami-nation(HE and Masson staining)to assess renal injury and fibrosis under light microscopy.Serum levels of urea nitrogen(Urea),serum creatinine(SCr),and β2-MG were measured using an automated biochemical analyzer.Serum ROS,SOD,and MDA levels were detected by ELISA.Immunohistochemistry(IHC)was used to detect the expression of α-SMA,TGF-β,FN,Collagen-I,SLC7A11,and GPX4 in renal tissues.Western blot(WB)was performed to analyze the protein expression of Keap1,Nrf2,and HO-1.The mRNA expression levels of Keap1,Nrf2,HO-1,SLC7A11,and GPX4 were quantified by real-time reverse transcription poly-merase chain reaction(RT-PCR).Results Compared with the sham-operated group,the model group exhibited significant pathological alterations,including marked renal tubular dilation and collagen deposition.The model group also showed increased levels of Urea,SCr,β2-MG,ROS,MDA,α-SMA,TGF-β,FN,Collagen-I,and Keap1 protein(P<0.05),alongside decreased levels of SOD,Nrf2 protein,HO-1 protein,GPX4 protein,and SLC7A11 protein(P<0.05).Drug interventions ameliorated the pathological changes and reduced fibrosis(P<0.05).Treated groups exhibited decreased levels of Urea,SCr,β2-MG,ROS,MDA,α-SMA,TGF-β,FN,Collagen-I,and Keap1 protein(P<0.05),and increased levels of SOD,Nrf2 protein,HO-1 protein,GPX4 protein,and SLC7A11 protein(P<0.05).The high-dose BHXF group demonstrated the most pronounced effects(P<0.05).Conclusion BHXF effectively attenuated renal fibrosis and improved renal function in CRF rats.Its underlying mechanism may involve regulating the Keap1/Nrf2/HO-1 signaling pathway to inhibit ferroptosis.