活性维生素D3对阿霉素肾病大鼠肾组织Akt磷酸化的影响  

Effects of active vitamin D3 on Akt phosphorylation in renal tissue of rats with adriamycin nephropathy

作　　者：陈亚茹 赵丹[2] 马国英 杨晓萍[2] 罗星[4] CHEN Ya-Ru;ZHAO Dan;MA Guo-Ying(Xinjiang Agricultural Vocational and Technical College,Changji 831100,Xinjiang,China)

机构地区：[1]新疆农业职业技术学院,新疆昌吉831100 [2]石河子大学医学院第一附属医院 [3]新疆维吾尔自治区人民医院 [4]石河子大学医学院

出　　处：《中国老年学杂志》2024年第14期3441-3447,共7页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(No.81160090);石河子大学科研项目(No.ZZZC201944A)。

摘　　要：目的研究活性维生素(AV)D3对阿霉素肾病(AN)大鼠肾组织蛋白激酶B(Akt)磷酸化(Ser473)的影响。方法将90只雄性SD大鼠均分为正常对照(NC)组、AN组及AVD3干预(AN+AVD3)组,每组各30只。除NC组外,其余均一次性尾静脉注射阿霉素7.5 mg/kg造模,造模成功后,AN+AVD3组0.25μg/(kg·d)AVD3灌胃治疗,其余两组给予等量的花生油灌胃,持续治疗10 w。采用免疫荧光(IF)观察p-Akt蛋白在肾小球中的定位;免疫组织化学染色(IHC)检测p-Akt蛋白的平均光密度;实时荧光定量聚合酶链反应(PCR)检测Akt mRNA水平;Western印迹检测Akt及p-Akt蛋白表达。结果造模大鼠精神、饮食、排泄都出现异常(大量蛋白尿),组织化学染色呈现严重的病理学损伤;IF结果显示:p-Akt蛋白定位于肾小球系膜基质区,连续样分布,AN组较NC组阳性信号明显增强,AN+AVD3组较AN组阳性信号减弱;IHC结果显示:实验后,AN组p-Akt蛋白的平均吸光度(第6周起)显著高于NC组,AN+AVD3组p-Akt蛋白的平均吸光度显著低于AN组(P<0.001);实时荧光定量PCR和Western印迹实验显示:实验后,各组Akt mRNA和蛋白水平未见明显差异(P>0.05),但AN组p-Akt/Akt较NC组显著升高,AN+AVD3组p-Akt/Akt较AN组显著降低(P<0.01,P<0.001)。结论AVD3显著抑制AN肾小球内Akt的磷酸化,提示AVD3可能通过抑制磷脂酰肌醇3-激酶(PI3K)/Akt通路发挥肾保护作用。Objective To investigate the effect of active vitamin(AV)D3 on protein kinase B(Akt)phosphorylation(Ser473)in renal tissue of adriamycin nephropathy(AN)rats.Methods A total of 90 male SD rats were divided into normal control(NC)group,AN group,and AVD3 intervention(AN+AVD3)group,with 30 rats in each group.Except for the NC group,the others were given a one-time tail vein injection of adriamycin 7.5 mg/kg to establish the model.After successful modeling,the AN+AVD3 group was given 0.25μg/(kg·d)AVD3 by gavage,and the other two groups were given the same amount of peanut oil by gavage,for a continuous treatment of 10 w.Immunofluorescence(IF)was used to observe the localization of p-Akt protein in the glomerulus;im-munohistochemical staining(IHC)was used to detect the average optical density of p-Akt protein;real-time fluorescence quantitative polymerase chain reaction(PCR)was used to detect the Akt mRNA level;Western blot was used to detect the expressions of Akt and p-Akt protein.Results The model rats showed abnormalities in mental,dietary and excretion(massive proteinuria),and histochemi-cal staining showed severe pathological damage;IF results showed that p-Akt protein was localized in the glomerular mesangial matrix area,with a continuous pattern distribution.The positive signal of the AN group was significantly stronger than that of the NC group,and the positive signal of the AN+AVD3 group was significantly weaker than that of the AN group;IHC results showed that after the ex-periment,the average absorbance of p-Akt protein(starting from the sixth week)in the AN group was significantly higher than that of the NC group,and the average absorbance of p-Akt protein in the AN+AVD3 group was significantly lower than that of the AN group(P<0.001);real-time fluorescence quantitative PCRand Western blot experiments showed that after the experiment,there was no sig-nificant difference in Akt mRNA and protein levels among each group(P>0.05),but the p-Akt/Akt in AN group was significantly higher than that in the NC group

关 键 词：活性维生素D3 阿霉素肾脏病 蛋白激酶B(Akt) P-AKT 

分 类 号：R692[医药卫生—泌尿科学]