核苷类似物对小鼠肝细胞线粒体DNA ND1和ND4的影响  

Mitochondrial DNA ND1 and ND4 mutation in liver and captured hepatocytes of mice after long-term nucleoside analogues exposure

作　　者：张维[1] 乔录新[1,2] 丁渭 陈德喜[1,2] 张玉林[1] ZHANG Wei;QIAO Luxin;DING Wei;CHEN Dexi;ZHANG Yulin(Beijing You′an Hospital,Capital Medical University,Beijing 100069,China;Beijing Institute of Hepatology,Beijing 100069,China)

机构地区：[1]首都医科大学附属北京佑安医院,北京100069 [2]北京市肝病研究所,北京100069

出　　处：《中国医药导报》2018年第21期8-13,共6页China Medical Herald

基　　金：国家自然科学基金资助项目(81571178;81371399)

摘　　要：目的探讨长期使用核苷类似物(NAs)是否导致小鼠肝脏线粒体DNA(mt DNA)ND1和ND4区损伤。方法 7周龄雌性Balb/c小鼠25只,采用简单随机分组法分为5组,对照组和4种核苷类似物组,每组各5只。实验组司他夫定(D4T)50 mg/kg,齐多夫定(AZT)100 mg/kg,拉米夫定(3TC)50 mg/kg和去羟肌苷(DDI)50 mg/kg,对照组为双蒸水,分别腹腔内注射,每周5次,连续3个月。留取各组肝组织,应用激光捕获显微技术获取肝细胞,对mt DNA ND1和ND4区克隆和测序。结果其余各组肝细胞的mt DNA ND4序列距离与参考序列的平均距离与对照组比较,差异均有高度统计学意义(P<0.01);AZT组肝细胞的mt DNA ND4平均d N与对照组比较,差异有统计学意义(P<0.05);肝细胞mt DNA ND1的序列距离与参考序列的平均距离,AZT组和3TC组序列距离与对照组比较,差异均有统计学意义(P<0.01);DDI组肝组织mt DNA ND1的平均d S与对照组比较,差异有统计学意义(P<0.05);AZT组和3TC组肝细胞mt DNA ND1的平均d S与对照组比较,差异均有统计学意义(P<0.05)。结论长期暴露于NAs可导致小鼠肝细胞mt DNA ND1和ND4区病变。Objective To investigate whether long-term use of nucleoside analogues(NAs)can lead to damage of DNA(mtDNA)ND1 and ND4 regions in liver mitochondria of mice.Methods According to the simple random method,twenty-five 7-weeks-old female Balb/c mice were divided into 5 groups,with 5 cases in each group.The control group received double-distilled water by intraperitoneal injection 5 days per week for 3 months.The each test group was respectively given D4T 50 mg/kg,AZT 100 mg/kg,3TC 50 mg/kg,or DDI 50 mg/kg by intraperitoneal injection per day,5 days per week for 3 months.Single liver cell was captured from mouse liver tissues by laser capture microdissection.The products of mtDNA ND1 and ND4 region PCR amplicons were cloned and sequenced in mice liver cells.Results The mean distance to the reference sequence of mtDNA ND4 in the captured hepatocytes of AZT,D4T and 3TC groups were longer than that in the control group,the differences were statistically significant(P<0.01).The mean mtDNA ND4 dN of the captured hepatocytes in the AZT group was higher than that in the control group,the difference was statistically significant(P<0.05).The mean distance to the reference sequence of mtDNA ND1 in the captured hepatocytes of AZT and 3TC groups were longer than that in the control group,the differences were statistically significant(P<0.01).The mean mtDNA ND1 dS of the liver tissue in the DDI group was higher than that in the control group,the difference was statistically significant(P<0.05).The mean mtDNA ND1 dS of AZT and 3TC groups were higher than that in the control group,the differences were statistically significant(P<0.05).Conclusion Long-term exposure to nucleoside analogue can result in mtDNA ND1 and ND4 regions lesion in mouse hepatocytes.

关 键 词：MTDNA ND1 MTDNA ND4 激光捕获显微切割 线粒体毒性 核苷类似物 

分 类 号：R978.7[医药卫生—药品] R512.91[医药卫生—药学]