本文關(guān)鍵詞： 激光掃描共聚焦顯微鏡 雙酚A 精母細(xì)胞 線粒體膜電位 熒光成像 熒光探針JC-　出處：《中國醫(yī)學(xué)裝備》2016年02期 　論文類型：期刊論文

【摘要】：目的:運(yùn)用激光掃描共聚焦顯微鏡(LSCM)技術(shù)檢測雙酚A(BPA)致精母細(xì)胞損傷后線粒體膜電位的變化,探討其作用機(jī)制。方法:將培養(yǎng)的GC-2小鼠精母細(xì)胞分為對照組、低劑量組和高劑量組,分別加入0、10μmol/L、100μmol/L的BPA,一同培養(yǎng)3 h后使用熒光探針JC-1對3個(gè)組樣本分別進(jìn)行標(biāo)記,采用LSCM檢測胞內(nèi)線粒體內(nèi)JC-1熒光強(qiáng)度的變化。利用LSCM專用軟件分析熒光強(qiáng)度,通過熒光顏色的轉(zhuǎn)變檢測線粒體膜電位的變化,用紅綠熒光的相對比例衡量線粒體去極化的比例。結(jié)果:對照組、低劑量組和高劑量組之間紅綠熒光的比值有著顯著差異。BPA低劑量組和高劑量組的胞內(nèi)線粒體膜電位明顯低于對照組,而高劑量組的胞內(nèi)線粒體膜電位較低劑量組低。結(jié)論:BPA對精母細(xì)胞有損傷,且隨著劑量加大,損傷越嚴(yán)重。LSCM技術(shù)方法靈敏度高,能夠?qū)崟r(shí)監(jiān)測細(xì)胞內(nèi)線粒體膜電位的變化。
[Abstract]:Objective: to investigate the changes of mitochondrial membrane potential (MMP) after spermatocyte injury induced by bisphenol A (BPA) by laser scanning confocal microscopy (LSCM). Methods: the cultured spermatocytes of GC-2 mice were divided into control group. In the low dose group and the high dose group, 10 渭 mol 路L ~ (-1) L ~ (-1) of 0 渭 mol / L ~ (100 渭 mol/L) was added respectively. The three groups were labeled with fluorescent probe JC-1 for 3 h. The changes of JC-1 fluorescence intensity in mitochondria were detected by LSCM, the fluorescence intensity was analyzed by LSCM software, and the change of mitochondrial membrane potential was detected by fluorescence color transformation. The ratio of mitochondrial depolarization was measured by the relative ratio of red and green fluorescence. Results: control group, There was significant difference in the ratio of red and green fluorescence between the low dose group and the high dose group. The intracellular mitochondrial membrane potential of BPA low dose group and high dose group was significantly lower than that of the control group. Conclusion: BPA has damage to spermatocytes, and with the increase of dose, the damage is more serious. LSCM technique has high sensitivity and can monitor the changes of mitochondrial membrane potential in the cells in real time. [WT5 "HZ] [WT5" BZ] [WT5BZ] [WT5 "BZ] [WT5" BZ] [WT5 "BZ]
【作者單位】： 第三軍醫(yī)大學(xué)生物醫(yī)學(xué)分析測試中心;
【分類號】：R114

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