异氟烷致新生大鼠睾丸生殖细胞凋亡及机制研究

发布时间：2018-04-25 20:40

  本文选题：异氟烷 + 凋亡　； 参考：《华中科技大学》2012年硕士论文

【摘要】：目的 探讨临床相关浓度的异氟烷对新生大鼠睾丸生殖细胞的影响及其作用机制。本课题为最终阐明吸入性全身麻醉药的生殖毒性及其作用机制，指导全身麻醉药的安全合理应用，以及探索其毒性作用的有效防治手段等方面提供了理论依据。 方法 60只出生1天（P1d)雄性大鼠随机分为对照组和实验组，对照组大鼠自主吸入空气，实验组大鼠则分别暴露于1MAC异氟烷+30%O2中2h、4h和6h（2h组、4h组及6h组）。运用TUNEL法对大鼠睾丸生殖细胞的凋亡进行评价；Western Blot法对大鼠睾丸生殖细胞Bcl-2、Bax、细胞色素C（Cytochrome C）和活化Caspase-3蛋白质表达水平进行检测；Real-Time PCR法对大鼠睾丸生殖细胞Bcl-2、Bax、Cytochrome C和Caspase-3mRNA表达水平进行测定。 结果 TUNE结果显示，对照组大鼠睾丸的生精小管内很少出现凋亡的阳性细胞，异氟烷暴露2h、4h、6h组均使大鼠睾丸的平均每生精小管内阳性细胞数显著增加。与对照组相比，实验组大鼠睾丸细胞Bcl-2蛋白水平表达下调（P0.05）；且Bcl-2蛋白的表达在6h组较4h组下调（P0.05）。4h和6h组与空白对照组相比较，大鼠睾丸细胞Bcl-2mRNA水平表达下调（P0.05）；Bcl-2mRNA的表达在2h、4h和6h依次下调（P0.05）。实验组大鼠的Bax的蛋白质表达水平较对照组上调（P0.05）；4h暴露组与2h暴露组相比，表达上调（P0.05）。4h、6h暴露组与空白对照组相比较，Bax mRNA表达水平明显上调（P0.05）；Bax的mRNA的表达水平在2h、4h和6h依次上调（P0.05）。与对照组比较，异氟烷暴露2h、4h、6h均使大鼠睾丸Cytochrome C蛋白表达增加（P0.05）。Cytochrome C mRNA的表达在4h和6h组表达上调（P0.05）。睾丸内活化Caspase-3蛋白及Caspase-3的mRNA表达量在实验组均明显升高，与空白对照组相比，差异有统计学意义（P0.05）；活化Caspase-3蛋白及Caspase-3mRNA的表达在2h组、4h组和6h组依次上调（P0.05）。 结论 通过对新生大鼠暴露于异氟烷的研究证实，，P1d大鼠暴露于1MAC的异氟烷2h将致睾丸生殖细胞凋亡，随着暴露时间延长至6h，其凋亡效应呈现时间依赖性。Bcl-2、Bax和Cytochrome C参与了异氟烷所致的新生大鼠生殖细胞凋亡。
[Abstract]:Purpose To investigate the effect of clinical concentration of isoflurane on testicular germ cells in neonatal rats and its mechanism. This paper provides a theoretical basis for elucidating the reproductive toxicity of inhaled general anesthetic and its mechanism, guiding the safe and rational application of general anesthetic, and exploring the effective prevention and treatment of its toxic effect. Method 60 male rats were randomly divided into two groups: control group and experimental group. Rats in control group inhaled air autonomously, rats in experimental group were exposed to 1MAC isoflurane 30%O2 for 2 h for 4 h, 6h(2h group for 4 h and 6h(2h group for 4 h and 6 h group, respectively. Evaluation of apoptosis of rat testicular germ cells by TUNEL method the expression of Bcl-2 C(Cytochrome C (cytochrome C) and activated Caspase-3 protein in rat testis germ cells were detected by Western Blot method. Real-time PCR method was used to detect the expression of Bcl-2Baxtochrome C and Caspase-3mRNA in rat testicular germ cells. Expression level was measured. Result TUNE results showed that there were few apoptotic positive cells in the seminiferous tubules of the control group, and the average number of positive cells per seminiferous tubule was significantly increased in the rats exposed to isoflurane for 2 h or 4 h or 6 h. Compared with the control group, the expression of Bcl-2 protein in testicular cells of the experimental group was down-regulated, and the expression of Bcl-2 protein was down-regulated in the 6h group compared with the 4h group and the 6h group compared with the blank control group. The expression of Bcl-2mRNA in rat testicular cells was down-regulated at 4 h and 6 h, respectively, and the expression of Bcl-2 mRNA was down-regulated at 2 h, 4 h and 6 h, respectively. The protein expression level of Bax in the experimental group was significantly higher than that in the control group at 4 h and 2 h, respectively. The expression of P0.05 mRNA was significantly up-regulated in the 6h group compared with the control group at 4 h and 6 h, respectively, and the expression level of mRNA of P0.05 was upregulated at 4 h and 6 h, respectively, in comparison with the control group. Compared with the control group, isoflurane increased the expression of Cytochrome C protein in testis of rats at 4 h and 6 h after exposure to isoflurane for 4 h and 6 h, respectively. The expression of Cytochrome C mRNA was up-regulated at 4 h and 6 h after exposure to isoflurane. The expression of activated Caspase-3 protein and mRNA of Caspase-3 in testis were significantly higher in the experimental group than in the blank control group, and the expression of activated Caspase-3 protein and Caspase-3mRNA were up-regulated in the groups of 2h and 6h, respectively. Conclusion Exposure to isoflurane in neonatal rats confirmed that isoflurane exposed to 1MAC for 2 hours induced apoptosis of germ cells in testis. As the exposure time was prolonged to 6 h, the apoptotic effect was time-dependent. Bcl-2 Bax and Cytochrome C were involved in the apoptosis of neonatal rat germ cells induced by isoflurane.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R726.1

【参考文献】

相关期刊论文 前1条

1 刘上峰;人类睾丸生殖细胞凋亡相关基因的分子遗传学研究进展[J];国外医学.遗传学分册;2002年03期

本文编号：1802879