Protective Effect And Mechanism Of Danhong Injection On Acute Kidney Injury Induced By Ischemia Reperfusion In Rats

Background and Objective: Acute kidney injury(AKI)is a clinical syndrome caused by a rapid decline in renal function with multiple potential causes,and is associated with high morbidity and mortality.Ischemia-reperfusion injury(IRI)is an imperative pathogenesis of clinical AKI.Preclinical studies have shown that AKI is associated with oxidative stress,apoptosis and inflammation.However,there is no specific treatment strategy for AKI.DHI protects against myocardial ischemia and cerebral ischemic diseases and these protective effects be related to its antiinflammatory action.Furthermore,DHI can treat kidney diseases such as diabetic nephropathy.In this study,we established an IRI rat model and a hypoxia/reoxygenation(H/R)injury cell culture model to mimic IRI in the kidney and explored the protective of DHI on ischemia-reperfusion(IR)-induced AKI.This study aimed to provide a valuable basis for the prevention and treatment of AKI and the development of innovative drugs.Methods: Protective effect of Danhong injection on acute kidney injury induced by ischemia reperfusion in rats:(1)The DHI group was administered a volume of 3 mL/(kg*d)DHI,and the sham group and model group were administered an equal volume of NaCl.(2)Established an IRI rat model.At the experimental endpoint,serum creatinine(Scr)and blood urea nitrogen(BUN)levels were measured.Periodic acid-schiff(PAS)staining was used to score renal pathology based on histopathologic changes.(3)A TUNEL assay was used to detect renal tubular cell apoptosis.Immunofluorescence was used to detect the infiltration of macrophages in vivo.Protective effect of Danhong injection on NRK-52 E cells injury induced by hypoxia/reoxygenation(H/R):(1)Cell viability was determined using a cell counting kit-8(CCK-8)assay for the identification of the appropriate DHI pretreatment concentration and duration.(2)Established a hypoxia/reoxygenation(H/R)injury cell culture model and measured cells apoptosis was examined by flow cytometry.(3)The protein expression of kidney injury molecule 1(KIM-1),tumor necrosis factor-α(TNF-α),nuclear factor κB P65(NF-κB P65)and phosphorylated(p)-NF-κB P65 was determined by Western blot in vivo and in vitro.(4)The mRNA expression of KIM-1,TNF-α and interleukin-6(IL-6)were analyzed by reverse transcription polymerase chain reaction in vivo and in vitro.Results: Treatment of IR rats with DHI markedly reduced Scr and BUN levels,attenuated renal pathological damage and alleviated macrophage infiltration in vivo.DHI effectively attenuated the apoptosis of renal tubular cells,decreased the expression of NF-κB signaling pathway related proteins such as KIM-1,TNF-α,NF-κB P65 and p-NF-κB P65,and reduced the release of inflammatory cytokines such as KIM-1,TNF-α and IL-6 in vivo and in vitro.Conclusion: DHI reduces kidney damage and macrophage infiltration induced by IR-AKI and apoptosis of NRK-52 E cells induced by H/R.At the same time,DHI reduces the expression level of KIM-1 and inhibits the NF-κB signaling pathway in vivo and vitro.DHI exhibits renoprotective effects against IRI-induced AKI in rats via the NF-κB signaling pathway.This study provides a new idea for the treatment of AKI via the target of NF-κB signaling pathway and provides a theoretical basis and reference for the application of DHI in clinical practice.