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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.
Renal anemia mice model was established by intragastric administration of adenine (50 mg · kg -1). The peripheral blood red blood cell count (RBC), reticulocyte count (Ret) And hemoglobin (Hb). ELISA kit was used to determine the content of erythropoietin (EPO) and stem cell factor (SCF). Serum iron, malondialdehyde (MDA) and superoxide dismutase (SOD) The model of iron overload chronic kidney injury was established by intraperitoneal injection of iron dextran (100 mg · kg -1), and the injury of renal tissue was evaluated by HE staining two weeks after the model was established. The levels of ROS, Tumor necrosis factor (TNFα), interleukin 6 (IL-6), hs-CRP and renal tissue 8-OHdG were determined by ELISA. Serum iron and total antioxidant capacity -AOC, MDA and total bilirubin were detected by immunohistochemical method.Western blot was used to detect the expression of HO-1 in kidneys.Results showed that in the model of renal anemia, Sheng-Ni-Ning (400,200,100 mg · kg ~ (-1) 1), ig) can reduce the level of RBC, Ret, Hb, increase serum iron, SCF content, reduce MDA content.In iron overload chronic kidney injury model, (400, 200mg · kg -1, ig) could improve the renal pathology; reduce serum iron, serum total bilirubin, TNFα, IL-6, hs-CRP, ROS, MDA and renal tissue 8-OHdG content; Improve the activity of T-AOC and up-regulate the expression of HO-1.The above results show that Shengxuening has significant preventive and therapeutic effects on renal anemia and iron overloaded chronic kidney injury mice and has a bidirectional regulation of iron in the body and can avoid iron overload oxidation Shock response, the mechanism may be related to the up-regulation of HO-1 protein expression.