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  本文選題：吡咯喹啉醌 + 氧糖剝奪/復(fù)氧　； 參考：《大連醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的:腦缺血后的病理生理過程復(fù)雜,腦缺血再灌注損傷級聯(lián)反應(yīng)其中一環(huán)是細(xì)胞凋亡,抑制神經(jīng)細(xì)胞凋亡有利于缺血性腦血管病的治療和康復(fù)。近期國內(nèi)外研究表明,吡咯喹啉醌(Pyrroloquinoline quinone ,PQQ)具有較強(qiáng)的抗氧化作用,可能通過抗氧化應(yīng)激對腦缺血有保護(hù)作用。而氧化應(yīng)激反應(yīng)被認(rèn)為是導(dǎo)致凋亡的最重要途徑之一,因此本文研究吡咯喹啉醌(PQQ)是否具有抗凋亡作用及其可能機(jī)制。研究吡咯喹啉醌(PQQ)對體外培養(yǎng)小鼠神經(jīng)母細(xì)胞瘤Neuro2A細(xì)胞株氧糖剝奪/復(fù)氧模型(OGD/R)的影響,在細(xì)胞水平探討PQQ神經(jīng)保護(hù)作用的可能機(jī)制。 方法:利用不同濃度PQQ預(yù)處理Neuro2A細(xì)胞后,通過無氧密閉盒和無糖培養(yǎng)液,建立氧糖剝奪模型。取復(fù)氧后6h這一時間點(diǎn),觀察PQQ干預(yù)對Neuro2A細(xì)胞的細(xì)胞形態(tài)、存活率、凋亡率以及氧化應(yīng)激檢測指標(biāo)活性氧(ROS)和還原型谷胱甘肽(GSH)的影響。細(xì)胞形態(tài)學(xué)采用倒置相差顯微鏡觀察;細(xì)胞存活率采用四唑鹽(MTT)法檢測;細(xì)胞凋亡率采用熒光顯微鏡觀察;ROS和GSH采用熒光分光光度計(jì)檢測。 結(jié)果:一.倒置相差顯微鏡觀察細(xì)胞形態(tài):OGD 2h復(fù)氧6h后Neuro2A細(xì)胞形態(tài)出現(xiàn)嚴(yán)重?fù)p傷,6.4μM和12.8μM的PQQ預(yù)處理有減輕神經(jīng)細(xì)胞損傷的作用;二.MTT法檢測細(xì)胞存活率:OGD 2h復(fù)氧6h后,0 12.8μM的PQQ預(yù)處理組細(xì)胞的存活率呈逐漸增高趨勢(P0.05)。三.熒光顯微鏡下計(jì)數(shù)細(xì)胞凋亡率:OGD 2h復(fù)氧6h后,6.4μM、12.8μM的PQQ預(yù)處理組和OGD無保護(hù)組相比細(xì)胞凋亡數(shù)逐漸減少(P0.05)。四.PQQ預(yù)處理后細(xì)胞內(nèi)ROS生成減少(P0.01),GSH水平增高(P0.05),與氧糖剝奪/復(fù)氧組相比有統(tǒng)計(jì)學(xué)差異。 結(jié)論:一.PQQ對神經(jīng)細(xì)胞氧糖剝奪損傷有一定的保護(hù)作用。二.PQQ對氧糖剝奪損傷的Neuro2A細(xì)胞的神經(jīng)保護(hù)作用可能是通過減輕氧化應(yīng)激反應(yīng),降低細(xì)胞凋亡率來實(shí)現(xiàn)的。
[Abstract]:Aim: the pathophysiological process after cerebral ischemia is complicated. One of the cascade reactions of cerebral ischemia reperfusion injury is apoptosis. Inhibition of neuronal apoptosis is beneficial to the treatment and rehabilitation of ischemic cerebrovascular disease. Recent studies at home and abroad have shown that pyrroloquinoline quinone (PQQ) has a strong antioxidant effect, which may have protective effect on cerebral ischemia through antioxidant stress. Oxidative stress reaction is considered to be one of the most important ways to induce apoptosis. Therefore, whether pyrroloquinoline quinone (PQQ) has anti-apoptotic effect and its possible mechanism is studied in this paper. To study the effect of pyrroloquinoline quinone (PQQ) on the oxygen deprivation / reoxygenation model (OGD / R) of mouse neuroblastoma cell line Neuro2A in vitro, and to explore the possible mechanism of PQQ neuroprotective effect at the cell level. Methods: Neuro2A cells were pretreated with different concentrations of PQQ. At 6 h after reoxygenation, the effects of PQQ on cell morphology, survival rate, apoptosis rate, reactive oxygen species (Ros) and reduced glutathione (GSH) in Neuro2A cells were observed. Cell morphology was observed by inverted phase contrast microscope, cell survival rate was detected by MTT assay, apoptosis rate was observed by fluorescence microscope and GSH was detected by fluorescence spectrophotometer. Results: 1. Observation of cell morphology by inverted phase contrast microscope (PQQ) after reoxygenation for 6 h after reoxygenation for 2 h at 1: 0GD could attenuate the injury of neuro2A cells by PQQ pretreatment of 6.4 渭 M and 12.8 渭 M, respectively. 2. MTT assay was used to detect the survival rate of cells in the PQQ pretreatment group with 12.8 渭 M of 0 渭 M after reoxygenation for 2 h and 6 h after reoxygenation for 2 h (P0.05). III. The apoptosis rate was counted under fluorescence microscope. The apoptosis rate of PQQ pretreatment group (6.4 渭 M) 12.8 渭 M after reoxygenation for 6 h after reoxygenation for 2 h was gradually decreased compared with that in OGD unprotected group (P0.05). 4. After pretreatment with PQQ, Ros production decreased (P0.01) and GSH level increased (P0.05), which was significantly different from that of glucose deprivation / reoxygenation group. Conclusion: 1. PQQ has protective effect on neuronal oxygen and glucose deprivation injury. 2. The neuroprotective effect of PQQ on neuro2A cells injured by oxygen glucose deprivation may be achieved by reducing oxidative stress response and apoptosis rate.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王霞,張璧濤,鐘樂,余小河,楊于嘉,陳光建;黃芩苷對Neuro2A細(xì)胞氧糖剝奪的保護(hù)作用[J];中國藥理學(xué)通報(bào);2005年10期

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本文編號：2056550