抗HBV药物富马酸替诺福韦二吡呋酯长期低剂量灌胃大鼠的肾脏毒性观察  

Renal toxicity in rats after long-term low-dose intragastric administration of anti-HBV drug TDF

作　　者：孔德亮 梅小斌 陈舟 徐爱静 KONG Deliang;MEI Xiaobin;CHEN Zhou;XU Aijing(Infection Department,The First Affiliated Hospital of Naval Medical University of the Chinese People's Liberation Army,Shanghai 200433,China)

机构地区：[1]中国人民解放军海军军医大学第一附属医院感染科,上海200433

出　　处：《山东医药》2024年第35期40-44,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(32070935)。

摘　　要：目的观察抗乙型肝炎病毒药物富马酸替诺福韦二吡呋酯(TDF)长期低剂量灌胃大鼠的肾脏损伤情况,以评价该药物的安全性。方法将30只SD大鼠随机分为空白对照组、生理盐水组、TDF-0w组、TDF-4w组、TDF-8w组,每组6只。TDF-0w组、TDF-4w组、TDF-8w组均给予TDF 1 mg/(kg·d)灌胃6周,停药时间分别为0、4、8周;生理盐水组给予等量生理盐水灌胃6周,空白对照组自由进食、进水6周。使用Sunrise酶标仪检测各组大鼠血磷、血钙、血β2微球蛋白(β2-MG)、血肌酐(Scr)及尿β2-MG、尿β-N-乙酰氨基葡萄糖苷酶(NAG),处死后取肾脏组织,采用HE染色及Massion染色观察肾脏组织病理情况,采用实时荧光定量PCR法检测肾脏组织肾损伤分子1(Kim-1)、阴离子转运蛋白1(Oat1)、阴离子转运蛋白2(Oat2)、多药耐药相关蛋白4(Mrp4)mRNA相对表达量,Western blotting法检测肾脏组织Kim-1、Mrp4蛋白相对表达量。结果各组血磷、血钙、血β2-MG、血Scr及尿β2-MG比较差异均无统计学意义(P均>0.05)。TDF-4w组大鼠尿NAG水平均低于其他各组(P均<0.05)。空白对照组和生理盐水组大鼠肾小球结构完整,肾小管上皮细胞排列整齐,肾脏基底膜完整且未出现病变;与空白对照组和生理盐水组比较,TDF-0w组大鼠部分肾小管上皮细胞存在轻度/中度肿胀变性和脱颗粒现象,有明显肾脏纤维化及炎症细胞浸润,但总体上肾小球结构完整,未观察到明显肾小管坏死现象;与TDF-0w组比较,TDF-4w组和TDF-8w组大鼠肾小球病理形态改善,肾脏纤维化及炎症细胞浸润均减轻。与空白对照组及生理盐水组比较,TDF-0w组、TDF-4w组大鼠肾脏组织Kim-1、Oat1、Oat2、Mrp4 mRNA及Kim-1、Mrp4蛋白相对表达量均升高,TDF-0w组、TDF-4w组、TDF-8w组上述指标依次降低(P均<0.05)。结论TDF长期低剂量灌胃大鼠会出现肾小管病理损伤,这种损伤会随着TDF停药时间的延长而改善。Objective To observe the renal injury in rats after long-term low-dose intragastric administration of teno-fovir disoproxil fumarate(TDF)in order to evaluate the safety of this drug.Methods Thirty SD rats were randomly di-vided into the control group,normal saline group,TDF-0w group,TDF-4w group,and TDF-8w group,with 6 rats in each group.Rats in the TDF-0w,TDF-4w,and TDF-8W groups were given TDF 1 mg/(kg·d)by gavage for 6 weeks,and the drug withdrawal time was 0,4,and 8 weeks,respectively.Rats in the normal saline group were given the same volume of normal saline by gavage for 6 weeks,and rats in the blank control group were allowed to eat and drink freely for 6 weeks.Blood phosphorus,blood calcium,bloodβ2-microglobulin(β2-MG),serum creatinine(Scr),urineβ2-MG and urineβ-N-acetylglucosaminidase(NAG)were detected by Sunrise microplate analyzer.After the rats were sacrificed,the kidney tis-sues were taken for HE staining and Massion staining to observe the pathological changes of kidney tissues.The relative ex-pression levels of kidney injury molecule 1(Kim-1)antibody,anion transporter 1(Oat1),anion transporter 2(Oat2)and multidrug resistanence-associated protein 4(Mrp4)mRNA in the renal tissues were detected by real-time fluorescence quantitative PCR.Western blotting was used to detect the relative expression of Kim-1 and Mrp4 protein in kidney tissues.Results There were no significant differences in serum phosphorus,serum calcium,serumβ2-MG,serum Scr or urineβ2-MG among these groups(all P>0.05).The urinary NAG water in the TDF-4w group was significantly lower than that in the blank control,saline,TDF-0w,and TDF-8w groups(all P<0.05).In the blank control group and the normal saline group,the glomerular structure was intact,the renal tubular epithelial cells were arranged neatly,and the renal basement membrane was intact without lesions.Compared with the blank control group and the saline group,the TDF-0w group showed mild/moderate swelling,degeneration and degranulation of some renal tubular epithelial cells

关 键 词：乙型肝炎病毒 核苷酸类似物 富马酸替诺福韦二吡呋酯 肾小管损伤 慢性肾损伤 

分 类 号：R33[医药卫生—人体生理学]