Protective Effects Of Huaiqihuang Extractum In Cisplatin-Induced Acute Kidney Injury

Objective Cisplatin plays an important role in the treatment of various solid tumors, but its severe renal toxicity limits its use. Previous study had showed that, cisplatin induces acute kidney injury via various mechanisms including tubular epithelial cell necrosis, apoptosis and inflammatory injury. In this study,by using a cisplatin induced acute kidney injury in mice, we observed Huaiqihuang extractum pretreatment effect on cisplatin induced renal tubular epithelial cell apoptosis, programmed necrosis. And, we further investigated the renal protective mechanism of Huaiqihuang extractum in cisplatin-induced acute kidney injury.Methods 1.The acute kidney injury in mice was induced by cisplatin. 25 g weight C57 BL / 6 male mice were divided into the following groups:(1) Con group : Intraperitoneal injection of saline(0.02 ml / g body weight), three days later the mice were sacrificed.(2) Cis15mg/kg group: Intraperitoneal injection of cisplatin(15mg/kg body weight), three days later the mice were sacrificed.(3)Cis20mg/kg group: Intraperitoneal injection of cisplatin(20mg/kg body weight),three days later the mice were sacrificed.(4) Cis25mg/kg group: Intraperitoneal injection of cisplatin(25mg/kg body weight), three days later the mice were sacrificed. Serum urea nitrogen and PAS staining of renal sections were analyzed to obtain the optimum cisplatin induced-AKI. 2. In the following experiments, mice were divided into four groups to investigate the renal protection of Huaiqihuang extractum pretreatment in cisplatin indcued-AKI.(1)Con group: In normal saline(0.02 ml / g body weight) two days and one hour before the injection given normal saline(0.015 ml / g body weight) gavage pretreatment.(2) Cis group: In cisplatin(20mg/kg body weight) two days and one hour before the injection given normal saline(0.015 ml / g body weight)gavage pretreatment.(3) HQH+Cis group:In cisplatin(20mg/kg body weight)two days and one hour before the injection given Huaiqihuang extractum(6g/kg body weight) gavage pretreatment.(4) Dex+Cis group: In intraperitoneal injection of cisplatin(20mg / kg body weight) one hour before dexamethasone(4mg / kg body weight) by intraperitoneal injection pretreatment. Serum urea nitrogen and PAS staining was analyzed to evaluate renal function and renal damage. Immunohistochemistry, immunofluorescence, PCR and western blot were used to analyze kidney inflammation and necrosis.Results We found that with the increase of the cisplatin concentration, serum urea nitrogen and kidney damage exacerbated in cisplatin treated-mice.(P<0.05). The optimum concentration of cisplatin induced-AKI in mice was20mg/kg, which was used for the following experiments. We found that casts or tubular epithelial cell injury, serum urea nitrogen were significantly reduced the both HQH+Cis group and Dex+Cis group, compared to Cis group(P <0.01).Acute kidney injury indicator Kim-1, programmed necrosis index RIP3,inflammatory cytokines TNF- α, IL-1β, HMGB1 karyoplasmic transfer and apoptosis index of TUNEL were found decreased in HQH+Cis group and Dex+Cis group. Western blot showed pro-apoptotic index of Bax decreased(P <0.06), and the anti-apoptotic index of BCL-2 increased(P <0.06) in HQH+Cis group and Dex+Cis group. Compared to Cis group, HQH+Cis group and Dex+Cis group demonstrated less.Conclusion Huaiqihuang extractum pretreatment ameliorates cisplatin induced AKI by inhibiting the renal inflammation, and renal tubular apoptosis and necrosis.