阿德福韦酯联合拉米夫定对大鼠肾细胞线粒体毒性的研究

发布时间：2018-11-13 17:47

【摘要】：目的评估阿德福韦酯(adefovir dipivoxil,ADV)和拉米夫定(lamivudine,LAM)联合应用对大鼠肾细胞线粒体的毒性损伤情况，为临床的安全用药提供参考。方法 1.动物分组及给药将40只清洁级SD大鼠随机分为LAM组、ADV组、ADV+LAM组和对照组，每组10只（雌雄各半），连续灌胃给药8周，其中LAM组给予LAM(300mg kg-1d-1)，ADV组给予ADV (40mg kg-1d-1)，，两药联用组给予ADV(40mg kg-1d-1)+LAM(300mg kg-1d-1)，对照组则给予生理盐水。 2.本实验主要的检测指标和方法 2.1采用全自动生化仪检测各组大鼠血清Cr、BUN水平，以评估大鼠肾脏的功能状态。 2.2采用透射电子显微镜观察SD大鼠肾细胞线粒体的超微形态结构变化，并采用Flameng方法进行分级评分，对线粒体的超微结构进行半定量分析并评分，以观察线粒体的功能状态和结构变化情况。 2.3采用实时荧光定量PCR (real time PCR)检测各组大鼠肾细胞线粒体DNA（mtDNA）的含量，线粒体DNA的含量与线粒体的氧化磷酸化功能密切相关。通过检测线粒体DNA水平可以反映其功能状态，同时也是反映线粒体基因损伤的指标之一。结果 1.各组大鼠血肌酐、尿素氮水平差异无统计学意义，P值分别为0.14和0.08。 2.LAM组、ADV组、ADV+LAM组大鼠肾细胞线粒体细胞色素b的基因含量分别为0.79±0.12、0.59±0.13、0.42±0.09，均低于对照组(1.00±0.00，F=63.72P0.05），其中ADV组较LAM组基因含量低，P0.05；ADV+LAM组分别与LAM及ADV两个单药组比较基因含量最低，P值均0.05。 3.透射电子显微镜下观察对照组、LAM组、ADV组及ADV+LAM组肾细胞线粒体评分分别为(0.19±0.02、0.28±0.03、0.43±0.04、0.56±0.07)，三个用药组较对照组损伤明显(F=80.38P0.05)，其中ADV组较LAM组损伤重，P0.05；ADV+LAM组与LAM及ADV两个单药组比较损伤最重，P值均0.05。结论 1.拉米夫定及阿德福韦酯均能对肾脏线粒体DNA聚合酶产生抑制作用，从而引起大鼠肾细胞线粒体损伤。 2.阿德福韦酯和拉米夫定两种药物联合应用对大鼠肾细胞线粒体的损伤具有叠加作用。 3.血肌酐、尿素氮对轻微肾脏损伤不能及时反映，故应进一步监测反映肾损伤较敏感的指标。
[Abstract]:Objective to evaluate the toxicity of adefovir dipivoxil (adefovir dipivoxil,ADV) combined with lamivudine (lamivudine,LAM) on mitochondria of renal cells in rats. Method 1. Forty clean SD rats were randomly divided into LAM group, ADV group and control group, 10 rats in each group (half of male and female), and the LAM group was given LAM (300mg kg-1d-1) for 8 weeks. ADV (40mg kg-1d-1) was given to ADV group, ADV (40mg kg-1d-1) LAM (300mg kg-1d-1) was given to two drugs combined group, and normal saline was given to control group. 2. The main indexes and methods 2. 1 the serum Cr,BUN level of rats in each group was measured by automatic biochemical instrument to evaluate the function of rat kidney. 2.2 the ultrastructural changes of mitochondria in renal cells of SD rats were observed by transmission electron microscope. The ultrastructure of mitochondria was graded by Flameng method, and the ultrastructure of mitochondria was evaluated by semi-quantitative analysis. The function and structure of mitochondria were observed. 2.3 the content of mitochondrial DNA (mtDNA) was measured by real-time fluorescence quantitative PCR (real time PCR). The content of mitochondrial DNA was closely related to the oxidative phosphorylation of mitochondria. The level of mitochondrial DNA can reflect the functional state of mitochondria, and it is also one of the indicators of mitochondrial gene damage. Result 1. There was no significant difference in serum creatinine and urea nitrogen levels among the three groups (P = 0.14 and 0.08, respectively). The mitochondrial cytochrome b gene content in 2.LAM group and ADV group was 0.79 卤0.120.59 卤0.130.42 卤0.09, respectively, which was lower than that in control group (1.00 卤0.00FN 63.72P0.05). The gene content in ADV group was lower than that in LAM group (P0.05). Compared with LAM and ADV, ADV LAM group had the lowest gene content (P < 0.05). 3. The mitochondria scores of renal cells in the control group, LAM group, ADV group and ADV LAM group were (0.19 卤0.02) 0.28 卤0.03 卤0.43 卤0.04 卤0.56 卤0.07 under transmission electron microscope, respectively. The damage of renal cells in the three groups was significantly higher than that in the control group (F=80.38P0.05). The damage in ADV group was more serious than that in LAM group (P0.05). The damage in ADV LAM group was the most serious than that in LAM and ADV group (P < 0.05). Conclusion 1. Lamivudine and adefovir ester could inhibit the mitochondrial DNA polymerase of kidney and induce mitochondria damage in rat renal cells. 2. The combination of adefovir and lamivudine has a superposition effect on mitochondria damage of rat renal cells. 3. The serum creatinine and urea nitrogen could not reflect the slight renal injury in time, so the sensitive indexes should be further monitored.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.62

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