骨化三醇对阿霉素肾病大鼠肾组织nephrin表达的影响

Effect of calcitriol on the expression of nephrin in renal tissue of rats with adriamycin-induced nephropathy

目的研究骨化三醇对阿霉素(ADR)肾病大鼠肾组织损伤及足细胞裂隙隔膜分子(nephrin)表达的影响。方法6周龄雄性SD大鼠分为:正常对照组(Control组),ADR肾病组,骨化三醇治疗组(ADR+VD组)。大鼠模型以单次尾静脉注射阿霉素(7.5 mg/kg),治疗组大鼠灌胃法服用0.25μg/(kg·d)骨化三醇。所有大鼠在处死前称重、留尿检测24 h尿蛋白(UP),留血检测血清白蛋白(Alb)、尿素氮(BUN)、肌酐(Cre);苏木素-伊红(HE)染色、马松三色染色(Masson)、透射电镜(TEM)观察肾病理学结构;免疫荧光观察nephrin的定位;免疫组织化学染色半定量检测nephrin的表达;Western印迹检测nephrin蛋白表达。结果与ADR肾病组比,ADR+VD组24 h UP、BUN、Cre、肾脏指数显著降低(P<0.05),Alb水平显著升高(P<0.05);组织化学染色显示骨化三醇减轻肾组织系膜基质增生及纤维化;TEM显示骨化三醇修复足细胞损伤;免疫组化与Western印迹检测结果显示高度一致性,骨化三醇升高ADR引起的nephrin表达降低(P<0.05)。结论骨化三醇能够减轻ADR引起的肾损伤和大量蛋白尿,这与改善肾足细胞标记分子nephrin低表达密切相关。

Objective To explore the effect of calcitriol on renal injury in rats with adriamyicin(ADR)nephropathy.Methods Male SD rats aged 6 weeks were divided into normal control(Control group),ADR nephropathy model(ADR nephropathy group),and calcitriol treatment groups(ADR+VD group).In the rat model,doxorubicin(7.5 mg/kg)was injected into the tail vein,and the rats in the treatment group were administered 0.25μg/(kg·d)calcitriol by intragastric administration.Before killed,all rats were weighted;24 h urine protein(UP),serum albumin(Alb),blood urea nitrogen(BUN),creatinine(Cre)were measured;Hematoxylin-Eosin(HE),Masson,transmission electron microscopy(TEM)were used to observe the structure of renal pathology;immunofluorescence was used to observe the localization of nephrin;immunohistochemical staining was used to detect the expression of nephrin semi-quantitatively;Western blot was used to detect the expression of nephrin protein.Results Compared with model group,calcitriol treatment significantly reduced 24 hUP,BUN,Cre,renal index(P<0.05),increased Alb level(P<0.05);histochemical staining showed calcitriol reduced renal mesangial matrix hyperplasia and fibrosis;TEM showed that calcitriol repaired podocyte injury;immunohistochemistry and Western blot showed high consistency,calcitriol increased the expression of nephrin induced by ADR(P<0.05).Conclusions Calcitriol could alleviate ADR-inducing kidney damage and massive proteinuria,which is closely related to the improvement of low expression of nephrin,a marker of renal foot cell marker.