【摘要】：目的 評估阿德福韋酯(adefovir dipivoxil,ADV)和拉米夫定(lamivudine,LAM)聯(lián)合應(yīng)用對大鼠腎細胞線粒體的毒性損傷情況，為臨床的安全用藥提供參考。 方法 1.動物分組及給藥 將40只清潔級SD大鼠隨機分為LAM組、ADV組、ADV+LAM組和對照組，每組10只（雌雄各半），連續(xù)灌胃給藥8周，其中LAM組給予LAM(300mg kg-1d-1)，ADV組給予ADV (40mg kg-1d-1)，，兩藥聯(lián)用組給予ADV(40mg kg-1d-1)+LAM(300mg kg-1d-1)，對照組則給予生理鹽水。 2.本實驗主要的檢測指標和方法 2.1采用全自動生化儀檢測各組大鼠血清Cr、BUN水平，以評估大鼠腎臟的功能狀態(tài)。 2.2采用透射電子顯微鏡觀察SD大鼠腎細胞線粒體的超微形態(tài)結(jié)構(gòu)變化，并采用Flameng方法進行分級評分，對線粒體的超微結(jié)構(gòu)進行半定量分析并評分，以觀察線粒體的功能狀態(tài)和結(jié)構(gòu)變化情況。 2.3采用實時熒光定量PCR (real time PCR)檢測各組大鼠腎細胞線粒體DNA（mtDNA）的含量，線粒體DNA的含量與線粒體的氧化磷酸化功能密切相關(guān)。通過檢測線粒體DNA水平可以反映其功能狀態(tài)，同時也是反映線粒體基因損傷的指標之一。 結(jié)果 1.各組大鼠血肌酐、尿素氮水平差異無統(tǒng)計學意義，P值分別為0.14和0.08。 2.LAM組、ADV組、ADV+LAM組大鼠腎細胞線粒體細胞色素b的基因含量分別為0.79±0.12、0.59±0.13、0.42±0.09，均低于對照組(1.00±0.00，F(xiàn)=63.72P0.05），其中ADV組較LAM組基因含量低，P0.05；ADV+LAM組分別與LAM及ADV兩個單藥組比較基因含量最低，P值均0.05。 3.透射電子顯微鏡下觀察對照組、LAM組、ADV組及ADV+LAM組腎細胞線粒體評分分別為(0.19±0.02、0.28±0.03、0.43±0.04、0.56±0.07)，三個用藥組較對照組損傷明顯(F=80.38P0.05)，其中ADV組較LAM組損傷重，P0.05；ADV+LAM組與LAM及ADV兩個單藥組比較損傷最重，P值均0.05。 結(jié)論 1.拉米夫定及阿德福韋酯均能對腎臟線粒體DNA聚合酶產(chǎn)生抑制作用，從而引起大鼠腎細胞線粒體損傷。 2.阿德福韋酯和拉米夫定兩種藥物聯(lián)合應(yīng)用對大鼠腎細胞線粒體的損傷具有疊加作用。 3.血肌酐、尿素氮對輕微腎臟損傷不能及時反映，故應(yīng)進一步監(jiān)測反映腎損傷較敏感的指標。
[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 鹵0.120.59 鹵0.130.42 鹵0.09, respectively, which was lower than that in control group (1.00 鹵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 鹵0.02) 0.28 鹵0.03 鹵0.43 鹵0.04 鹵0.56 鹵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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R512.62

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