益肾健脾化瘀汤通过调节Keap1/Nrf2/HO-1通路减轻STZ致大鼠肾损伤

Yishen Jianpi Huayu Decoction mitigated STZ induced kidney injury in rats by regulating the Keap1/Nrf2/HO-1 pathway

目的:基于Keap1/Nrf2/HO-1通路观察益肾健脾化瘀汤(YJHD)减轻链脲佐菌素(STZ)所致大鼠肾损伤的情况。方法:在60只SD雄性大鼠中随机选取10只作为对照组,普通维持饲料喂养,其余50只大鼠给予高脂饲料喂养,4 w后,单次小剂量腹腔注射STZ,将造模成功的40只大鼠按照每组10只随机分为模型组、YJHD低、高剂量组和阳性药物组[卡格列净(CANA)组],阳性药物选择卡格列净(CANA),灌胃8 w,同时间内模型组灌胃等容积的蒸馏水。给药后测定口服糖耐量(OGTT)以及曲线下面积(AUC),收集尿液测定24 h尿蛋白,取材时取血,用血清样本测定血肌酐(Cre)、血尿素氮(BUN)及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)及丙二醛(MDA)的水平,HE染色观察肾组织病理改变情况,Western blot迹检测肾组织中Keap1、Nrf2、HO-1水平。结果:与对照组相比,模型组FBG、AUC明显升高(P<0.05),24 h尿蛋白、Cre、BUN水平明显升高(P<0.05),MDA明显升高,SOD、GSH-Px以及CAT明显下降(P<0.05),肾组织中可见大量的炎性细胞增生浸润,Keap1的表达明显升高,Nrf2和HO-1的表达明显降低(P<0.05)。与阳性药物组相比,YJHD组FBG、AUC水平明显下降(P<0.05),24 h尿蛋白、Cre、BUN水平都有明显改善(P<0.05),MDA水平降低,SOD、GSH-Px、CAT水平提高(P<0.05),而在肾组织中,细胞的增生浸润情况有所缓解,Keap1的蛋白表达量降低,Nrf2和HO-1蛋白表达量明显升高(P<0.05)。结论:益肾健脾化瘀汤可能是通过Keap1/Nrf2/HO-1通路来缓解减轻STZ所致的大鼠的肾损伤。

Objective To observe the reduction of STZ-induced kidney injury in rats by Yishen Jianpi Huayu Decoction(YJHD)based on Keap1/Nrf2/HO-1 pathway.Method Ten of 60 SD male rats were randomly selected as the control group and fed with normal maintenance chow,while the remaining 50 rats were fed with high-fat chow,and after 4 weeks,a single low-dose intraperitoneal injection of streptozotocin(STZ)was administered,40 successfully modeled rats were randomly divided into 4 groups according to 10 rats in each group:model group,low group,high dose administration group and positive drug group[Cagliflozin(CANA)group],and the positive drug was chosen as Cagliflozin(CANA),which was gavaged for 8 weeks,while the model group was gavaged with an equal volume of distilled water during the same time.Oral glucose tolerance(OGTT)as well as area under the curve(AUC)were measured after drug administration,urine was collected to measure 24-hour urine protein,blood was taken at the time of sampling,and serum samples were used to measure blood creatinine(Cre),blood urea nitrogen(BUN),and levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT),and malondialdehyde(MDA).HE staining was performed to observe the pathological changes in renal tissues,and Western blot was performed to detect the levels of Keap1,Nrf2,and HO-1 in renal tissues.Results Compared with the control group,FBG and AUC were significantly increased(P<0.05),24-hour urine protein,Cre and BUN levels were significantly increased(P<0.05),MDA was increased,SOD,GSH-Px and CAT were decreased(P<0.05),a large number of inflammatory cell proliferation infiltration was seen in kidney tissues,and the expression of Keap1 was increased,Nrf2 and HO-1 expression decreased(P<0.05);compared with the positive drug group,FBG and AUC levels of YJHD group were seen to decrease(P<0.05),24-hour urine protein,Cre and BUN levels were significantly improved(P<0.05),MDA levels decreased,and SOD,GSH-Px and CAT levels increased(P<0.05),while in the renal tissue,the cell proliferation infiltration was alleviated,the protein expression of Keap1 was reduced,and the protein expression of Nrf2 and HO-1 was significantly increased(P<0.05).Conclusion Yishen Jianpi Huayu Decoction(YJHD)may alleviate and mitigate STZ-induced kidney injury in rats through the Keap1/Nrf2/HO-1 pathway.