發(fā)布時(shí)間：2018-01-18 15:26

  本文關(guān)鍵詞：線粒體在PBDE-47致SH-SY5Y細(xì)胞凋亡中的作用　出處：《華中科技大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： PBDE-47 SH-SY5Y細(xì)胞 細(xì)胞凋亡 線粒體膜電位 [Ca~(2+)]m 細(xì)胞色素C

【摘要】：多溴聯(lián)苯醚（Polybrominated diphenyl ethers，PBDEs）的神經(jīng)毒性作用近年來(lái)受到人們的廣泛關(guān)注，但其具體機(jī)制還不十分清楚。本實(shí)驗(yàn)通過(guò)PBDE-47作用于人神經(jīng)母細(xì)胞瘤細(xì)胞（SH-SY5Y細(xì)胞），觀察其對(duì)細(xì)胞凋亡率、線粒體膜電位（ΔΨm）、線粒體Ca~(2+)濃度（[Ca~(2+)]m）以及細(xì)胞色素C的影響，探討線粒體在PBDE-47致SH-SY5Y細(xì)胞凋亡中的作用，為PBDEs的神經(jīng)毒性作用機(jī)制研究提供理論依據(jù)。 第一部分PBDE-47對(duì)SH-SY5Y細(xì)胞凋亡率、線粒體膜電位和線粒體Ca~(2+)濃度的影響 目的：研究不同劑量PBDE-47對(duì)體外培養(yǎng)的SH-SY5Y細(xì)胞凋亡率、線粒體膜電位和線粒體Ca~(2+)濃度的影響。 方法：利用線粒體膜電位（JC-1）和線粒體鈣離子（Rhod-2/AM）檢測(cè)熒光探針處理細(xì)胞，分別采用流式細(xì)胞術(shù)（Flow cytometry,FCM）和熒光酶標(biāo)儀檢測(cè)PBDE-47作用對(duì)SH-SY5Y細(xì)胞凋亡率、ΔΨm和[Ca~(2+)]m的影響。 結(jié)果：與溶劑對(duì)照組比較，10μmol/L PBDE-47染毒組的細(xì)胞早期凋亡率、總凋亡率明顯升高（P 0.05）；10μmol/L PBDE-47染毒組ΔΨm和[Ca~(2+)]m顯著下降（P 0.05）。 結(jié)論：PBDE-47導(dǎo)致ΔΨm和[Ca~(2+)]m下降，可能與其誘導(dǎo)細(xì)胞凋亡相關(guān)。 第二部分PBDE-47對(duì)SH-SY5Y細(xì)胞內(nèi)細(xì)胞色素C mRNA和蛋白表達(dá)的影響 目的：探討PBDE-47對(duì)細(xì)胞色素C mRNA和蛋白表達(dá)水平的影響。 方法：提取胞漿RNA利用逆轉(zhuǎn)錄PCR法檢測(cè)胞漿細(xì)胞色素C的mRNA表達(dá)水平；分離、提取蛋白后利用Western blot技術(shù)檢測(cè)細(xì)胞胞漿和線粒體細(xì)胞色素C的蛋白表達(dá)水平。 結(jié)果：與溶劑對(duì)照組相比，10μmol/L PBDE-47染毒組胞漿細(xì)胞色素C mRNA表達(dá)水平明顯升高（P 0.05）；1、5、10μmol/L PBDE-47染毒組的線粒體細(xì)胞色素C蛋白表達(dá)水平顯著下降（P 0.05），同時(shí)，5、10μmol/L PBDE-47染毒組胞漿內(nèi)細(xì)胞色素C蛋白表達(dá)水平顯著升高（P 0.05）。結(jié)論：PBDE-47可導(dǎo)致細(xì)胞色素C從線粒體轉(zhuǎn)移到胞漿，從而激活線粒體信號(hào)通路，，誘導(dǎo)細(xì)胞凋亡。
[Abstract]:The neurotoxicity of polybrominated diphenyl ethersm (PBDEs) has attracted much attention in recent years. However, the specific mechanism is not very clear. In this experiment, human neuroblastoma cell line SH-SY5Y cells were treated by PBDE-47, and the apoptosis rate of SH-SY5Y cells was observed. Mitochondrial membrane potential, mitochondrial Ca~(2 concentration. [To explore the role of mitochondria in the apoptosis of SH-SY5Y cells induced by PBDE-47. To provide theoretical basis for the study of neurotoxicity mechanism of PBDEs. Effect of PBDE-47 on apoptosis rate, mitochondrial membrane potential and mitochondrial Ca~(2 concentration of SH-SY5Y cells Aim: to study the effects of different doses of PBDE-47 on apoptosis rate, mitochondrial membrane potential and mitochondrial Ca~(2 concentration in cultured SH-SY5Y cells. Methods: mitochondrial membrane potential (JC-1) and mitochondrial calcium ion Rhod-2 / AM were used to detect the fluorescence probe. Flow cytometry (FCM) and fluorescence enzyme labeling (FCM) were used to detect the apoptosis rate, 螖 蠄 m, and fluorescence enzyme of SH-SY5Y cells induced by PBDE-47. [Ca~(2)] m. Results: compared with the solvent control group, the early apoptosis rate and the total apoptosis rate in 10 渭 mol/L PBDE-47 group were significantly higher than those in the control group (P 0.05). 螖 蠄 m and 螖 蠄 m in 10 渭 mol/L PBDE-47 exposed group [Ca~(2)] m decreased significantly (P 0.05). Conclusion\\\% PBDE-47 leads to 螖 蠄 m sum. [The decrease of Ca~(2] m may be related to the induction of apoptosis. The effect of PBDE-47 on the expression of cytochrome C mRNA and protein in SH-SY5Y cells Objective: to investigate the effect of PBDE-47 on the expression of cytochrome C mRNA and protein. Methods: the mRNA expression level of cytochrome C was detected by reverse transcription PCR method. The protein expression of cytoplasmic and mitochondrial cytochrome C was detected by Western blot. Results: compared with the solvent control group, the expression of cytochrome C mRNA in 10 渭 mol/L PBDE-47 group was significantly higher than that in the control group (P 0.05). The expression of cytochrome C protein in mitochondria of 10 渭 mol/L PBDE-47 group decreased significantly (P 0.05). Cytochrome C protein expression in cytoplasm of 10 渭 mol/L PBDE-47 group increased significantly (P 0.05). Conclusion\% PBDE-47 can induce cytochrome C transfer from mitochondria to cytoplasm. Thus, mitochondrial signaling pathway was activated and apoptosis was induced.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R363

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相關(guān)期刊論文 前1條

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