發(fā)布時間：2018-04-25 20:40

  本文選題：異氟烷 + 凋亡　； 參考：《華中科技大學(xué)》2012年碩士論文

【摘要】：目的 探討臨床相關(guān)濃度的異氟烷對新生大鼠睪丸生殖細(xì)胞的影響及其作用機(jī)制。本課題為最終闡明吸入性全身麻醉藥的生殖毒性及其作用機(jī)制，指導(dǎo)全身麻醉藥的安全合理應(yīng)用，以及探索其毒性作用的有效防治手段等方面提供了理論依據(jù)。 方法 60只出生1天（P1d)雄性大鼠隨機(jī)分為對照組和實驗組，對照組大鼠自主吸入空氣，實驗組大鼠則分別暴露于1MAC異氟烷+30%O2中2h、4h和6h（2h組、4h組及6h組）。運用TUNEL法對大鼠睪丸生殖細(xì)胞的凋亡進(jìn)行評價；Western Blot法對大鼠睪丸生殖細(xì)胞Bcl-2、Bax、細(xì)胞色素C（Cytochrome C）和活化Caspase-3蛋白質(zhì)表達(dá)水平進(jìn)行檢測；Real-Time PCR法對大鼠睪丸生殖細(xì)胞Bcl-2、Bax、Cytochrome C和Caspase-3mRNA表達(dá)水平進(jìn)行測定。 結(jié)果 TUNE結(jié)果顯示，對照組大鼠睪丸的生精小管內(nèi)很少出現(xiàn)凋亡的陽性細(xì)胞，異氟烷暴露2h、4h、6h組均使大鼠睪丸的平均每生精小管內(nèi)陽性細(xì)胞數(shù)顯著增加。與對照組相比，實驗組大鼠睪丸細(xì)胞Bcl-2蛋白水平表達(dá)下調(diào)（P0.05）；且Bcl-2蛋白的表達(dá)在6h組較4h組下調(diào)（P0.05）。4h和6h組與空白對照組相比較，大鼠睪丸細(xì)胞Bcl-2mRNA水平表達(dá)下調(diào)（P0.05）；Bcl-2mRNA的表達(dá)在2h、4h和6h依次下調(diào)（P0.05）。實驗組大鼠的Bax的蛋白質(zhì)表達(dá)水平較對照組上調(diào)（P0.05）；4h暴露組與2h暴露組相比，表達(dá)上調(diào)（P0.05）。4h、6h暴露組與空白對照組相比較，Bax mRNA表達(dá)水平明顯上調(diào)（P0.05）；Bax的mRNA的表達(dá)水平在2h、4h和6h依次上調(diào)（P0.05）。與對照組比較，異氟烷暴露2h、4h、6h均使大鼠睪丸Cytochrome C蛋白表達(dá)增加（P0.05）。Cytochrome C mRNA的表達(dá)在4h和6h組表達(dá)上調(diào)（P0.05）。睪丸內(nèi)活化Caspase-3蛋白及Caspase-3的mRNA表達(dá)量在實驗組均明顯升高，與空白對照組相比，差異有統(tǒng)計學(xué)意義（P0.05）；活化Caspase-3蛋白及Caspase-3mRNA的表達(dá)在2h組、4h組和6h組依次上調(diào)（P0.05）。 結(jié)論 通過對新生大鼠暴露于異氟烷的研究證實，，P1d大鼠暴露于1MAC的異氟烷2h將致睪丸生殖細(xì)胞凋亡，隨著暴露時間延長至6h，其凋亡效應(yīng)呈現(xiàn)時間依賴性。Bcl-2、Bax和Cytochrome C參與了異氟烷所致的新生大鼠生殖細(xì)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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R726.1

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相關(guān)期刊論文 前1條

1 劉上峰;人類睪丸生殖細(xì)胞凋亡相關(guān)基因的分子遺傳學(xué)研究進(jìn)展[J];國外醫(yī)學(xué).遺傳學(xué)分冊;2002年03期

本文編號：1802879