生血宁对肾性贫血和铁过载慢性肾损伤小鼠的防治作用

Prevention and Treatment of Shengxuening on Renal Anemia and Chronic Renal Injury of Iron Overload in Mice

采用腺嘌呤(50mg·kg^(-1))灌胃的方式建立了肾性贫血小鼠模型,造模结束3周后测定小鼠外周血红细胞计数(RBC)、网织红细胞计数(Ret)和血红蛋白(Hb)含量;ELISA试剂盒测定促红细胞生成素(EPO)、干细胞因子(SCF)含量;商业化试剂盒测定血清铁、丙二醛(MDA)和超氧化物歧化酶(SOD)水平.采用右旋糖酐铁(100 mg·kg^(-1))腹腔注射的方法建立小鼠铁过载慢性肾损伤模型,造模结束2周后,HE染色评价肾组织损伤情况;ELISA试剂盒测定ROS、肿瘤细胞坏死因子(TNFα)、白细胞介素6(IL-6)、超敏C反应蛋白(hs-CRP)、肾组织8-OHdG含量;商业化试剂盒测定血清铁、总抗氧化能力(T-AOC)、MDA、总胆红素水平;Western检测肾脏血红素氧合酶-1(HO-1)蛋白表达.结果显示,在肾性贫血模型中,生血宁(400,200,100 mg·kg^(-1),ig)均可改善降低的RBC、Ret、Hb水平;增加血清铁、SCF含量,降低MDA含量.在铁过载慢性肾损伤模型中,生血宁(400,200mg·kg^(-1),ig)可改善肾脏病理;降低血清铁、血清总胆红素、TNFα、IL-6、hs-CRP、ROS、MDA和肾组织8-OHdG含量;提高T-AOC活性;上调HO-1表达.以上结果表明,生血宁对肾性贫血和铁过载慢性肾损伤小鼠具有明显的防治作用,对体内铁具有双向调节作用,能够避免铁过载氧化应激反应,其机制可能与上调HO-1蛋白表达有关.

The renal anemia model was induced by intragastric administration with adenine（50 mg·kg-1）in mice.Three weeks after the model was established,the peripheral blood RBC,Ret and Hb were determined.EPO and SCF were examined by ELISA.Commercial kits were used to measure serum levels of iron,MDA and SOD.The chronic renal injury model of iron overload was induced by intraperitoneal injection with iron dextran（100 mg·kg-1）in mice.Two weeks after the model was established,kidney tissue injury was evaluated by HE staining.ELISA was used to examine serum levels of ROS,TNFα,IL-6and hs-CRP and kidney content of 8-OHdG.Commercial kits were used to measure serum levels of iron,T-AOC,MDA and total bilirubin.Western blot was used to examine HO-1expression.Results：In the renal anemia model,Shengxuening（400,200,100mg·kg-1,ig）markedly improved decreased levels of RBC,Ret and Hb,increased serum levels of iron and SCF,decreased MDA content.In chronic renal injury model of iron overload,Shengxuening（400,200mg·kg-1,ig）decreased serum levels of iron,total bilirubin,TNFα,IL-6,hs-CRP,ROS and MDA and kidney content of 8-OHdG,promoted T-AOC activity and HO-1protein expression.In summary,shengxuening has obvious prevention and treatment effect on renal anemia and chronic renal injury of iron overload in mice with a bidirectional regulation effect on system iron which can avoid iron overload-induced oxidative stress,and its mechanism may be related to upregulation of HO-1protein expression.