基于Hepcidin/OPG-RANKL轴探讨当归多糖改善大鼠肾性贫血线粒体功能异常的作用机制

Mechanism Study of Angelica sinensis Polysaccharide on the Improvement of the Abnormal Mitochondrial Function in Renal Anemia Rats based on the Hepcidin/OPG-RANKL

目的:研究当归多糖对肾性贫血大鼠线粒体功能异常的影响及作用机制。方法:50只Wistar大鼠随机分为正常对照组、模型组及当归多糖低(100 mg/kg)、中(200 mg/kg)、高(400 mg/kg)剂量组,除正常对照组外,其余各组大鼠均采用腺嘌呤注射制备肾性贫血大鼠模型;证实成模后各组小鼠给予相应药物,正常对照组与模型组给予相应体积生理盐水,1次/d,连续给药14 d。检测各组大鼠外周血红细胞(RBC)计数、血红蛋白(Hb)水平、红细胞压积(Hct)、血尿素氮(BUN)、血清肌酐(Scr)、肾脏促红细胞生成素(EPO)、骨髓单个核细胞(BMMNCs)计数;通过流式细胞仪检测线粒体膜电位(MMP);RT-PCR检测骨髓单个核细胞Hepcidin、OPG、RANKL mRNA表达;Western blot检测骨髓单个核细胞Hepcidin、OPG、RANKL蛋白表达。结果:与正常对照组比较,模型组大鼠RBC、Hb、Hct、EPO水平及BMMNCs计数、BMMNCs线粒体膜电位、OPG mRNA及蛋白表达、OPG/RANKL比值显著降低,Scr、BUN水平和Hepcidin、RANKL mRNA及蛋白表达显著升高(P<0.01)。与模型组比较,当归多糖各剂量组大鼠RBC、Hb、Hct、EPO水平和BMMNCs线粒体膜电位均显著升高,Scr、BUN水平显著降低,当归多糖中、高剂量组BMMNCs计数、OPG mRNA及蛋白表达、OPG/RANKL比值显著升高,Hepcidin、RANKL mRNA及蛋白表达显著降低(P<0.01)。结论:当归多糖可有效治疗肾性贫血并改善其线粒体功能异常,其作用机制可能与调节Hepcidin/OPG-RANKL轴有关。

Objective:To study the effects and mechanism of Angelica sinensis polysaccharide(ASP)on mitochondrial dysfunction in renal anemia rats.Methods:50 Wistar rats were randomly divided into normal control group,model group,ASP low-,middle-and high-dose(100,200,400 mg/kg)groups.In addition to the normal control group,adenine injection was used to prepare renal anemia rat model.After confirmation of the formation of the model,rats in each group were given the corresponding drugs,the rats in normal control group and model group were given the corresponding volume of normal saline,1 time/d,continuously for 14 days.Peripheral blood red blood cell(RBC)count,hemoglobin(Hb)level,hematocrit(Hct),blood urea nitrogen(BUN),serum creatinine(Scr),renal erythropoietin(EPO),bone marrow mononuclear cell(BMMNCs)count were measured.Mitochondrial membrane potential(MMP)was detected by flow cytometry.The mRNA expressions of Hepcidin,OPG,RANKL in bone marrow mononuclear cells were detected by RT-PCR.Western blot was used to detect the expressions of Hepcidin,OPG,RANKL proteins in bone marrow mononuclear cells.Results:Compared with the normal control group,the levels of RBC,Hb,Hct and EPO,BMMNCs count,BMMNCs mitochondrial membrane potential,OPG mRNA and protein expressions,OPG/RANKL ratio of rats in the model group were significantly reduced,Scr,BUN levels,and the expressions of Hepcidin,RANKL mRNA and protein were significantly increased(P<0.01).Compared with the model group,the levels of RBC,Hb,Hct and EPO,BMMNCs mitochondrial membrane potential were significantly increased and the levels of Scr and BUN were significantly decreased in each dose group of ASP,BMMNCs count,OPG mRNA and protein expressions and OPG/RANKL ratio were significantly increased in the middle-and high-dose groups of ASP,the mRNA and protein expressions of Hepcidin,RANKL were significantly decreased(P<0.01).Conclusion:ASP can effectively treat renal anemia and improve its mitochondrial dysfunction,which may be related to regulating the Hepcidin/OPG-RANKL.