【摘要】：目的利用維生素K3(VK3)復制氧化應激模型,探討VK3誘導的ROS在Hela細胞溶酶體-線粒體細胞死亡途徑過程中的作用及機制。方法以人宮頸癌Hela細胞為研究對象,實驗分為:對照組、VK3組(30μmol/L)、NAC組(80μmol/L)和VK3與NAC聯(lián)合作用組。MTT法檢測細胞生存率,DCFH-DA染色觀察細胞內活性氧簇(ROS)水平,吖啶橙(AO)染色觀察溶酶體膜通透性變化,BCECF-AM染色流式細胞術檢測細胞質p H變化,羅丹明123(Rhodamine 123)染色檢測線粒體膜通透性變化。結果與對照組相比,單獨應用NAC對Hela細胞無明顯影響。VK3作用組Hela細胞生存率降低(P0.05),DCFH-DA染色熒光增強(P0.05),ROS水平增加;AO染色在VK3作用3 h后Hela細胞的細胞質綠色熒光增強(P0.05),紅色顆粒狀熒光減弱(P0.05),溶酶體通透性增加;BCECF-AM染色,VK3作用6 h后FL1/FL2比值降低(P0.05),細胞質呈酸性,酸性p H的細胞數(shù)為13.6%(P0.05);Rhodamin染色,VK3作用6 h后細胞質內紅色熒光強度降低(P0.05),線粒體膜電勢降低。與單獨應用VK3組比較,VK3與NAC聯(lián)合作用于Hela細胞時,細胞生存率增加(P0.05),ROS水平降低(P0.05),溶酶體通透性降低(P0.05),FL1/FL2比值增加(P0.05),酸性p H的細胞數(shù)為5.4%(P0.05),線粒體膜電勢增加(P0.05)。結論在氧化應激誘導Hela細胞損傷過程中,VK3通過ROS依賴途徑,誘導先后發(fā)生溶酶體及線粒體通透性增加,在這個過程中細胞質發(fā)生酸化。
[Abstract]:Objective to investigate the role and mechanism of VK3 induced ROS in the process of lysosomal mitochondrial cell death in Hela cells by using vitamin K3 (VK3) to replicate oxidative stress model. Methods Human cervical cancer Hela cells were divided into two groups: control group (30 渭 mol/L), control group (30 渭 mol/L) and VK3 combined with NAC group. Changes in membrane permeability of lysosome were observed by (AO) staining with acridine orange. The changes of cytosolic pH were detected by flow cytometry with BCECF-AM staining, and mitochondrial membrane permeability by Rhodamine 123 (Rhodamine 123) staining. Results compared with the control group, The survival rate of Hela cells in the Hela cells treated with NAC alone decreased (P0.05) the fluorescence intensity of DCFH-DA staining increased (P0.05) the cytoplasmic green fluorescence of Hela cells increased after 3 h of VK3 treatment (P0.05), and the red granular fluorescence decreased (P0.05). Weak (P0.05), the permeability of lysosome increased, the ratio of FL1/FL2 decreased after 6 h exposure to BCECF-AM staining (P0.05), and the cytoplasm was acidic. The number of acidic pH cells was 13.6% (P0.05). The red fluorescence intensity in cytoplasm was decreased (P0.05) and the mitochondrial membrane potential was decreased after VK3 was treated with Rhodamin staining for 6 h. Compared with the VK3 group, the cell survival rate increased (P0.05), the lysosomal permeability decreased (P0.05), the ratio of FL1 / FL2 increased (P0.05), the number of acidic pH cells increased 5.4% (P0.05), and the mitochondrial membrane potential increased (P0.05). Conclusion during oxidative stress induced Hela cell injury, VK3 induces the increase of lysosome and mitochondrial permeability through ROS dependent pathway, and the cytoplasm acidification occurs during this process.
【作者單位】： 北華大學基礎醫(yī)學院;國家食品藥品監(jiān)督管理總局藥品審評中心;
【基金】：吉林省教育廳資助課題〔吉教科合字2011第117號;2009第156號〕 吉林省科技廳自然科學基金資助課題〔201215103〕 吉林省衛(wèi)計委資助課題〔2013Z062〕 吉林市科技局項目〔201464064〕
【分類號】：R363

【相似文獻】

相關期刊論文 前5條

1 時艷艷;岳麓;齊衛(wèi)衛(wèi);魏紅梅;王秀美;邱文生;;消癌平治療小鼠惡性腹水的實驗研究[J];中華腫瘤防治雜志;2011年09期

2 ;顱腦[J];中國醫(yī)學文摘(腫瘤學);2005年02期

3 張平;趙梅;臧紅彥;鄒菁;葉欣;;50℃熱療對CT-26細胞體外作用的觀察[J];中華腫瘤防治雜志;2012年15期

4 吳偉忠,劉康達,湯釗猷,湯曉雷,吳厚生,胡新美,謝倩,高艷琴;熱休克對H22細胞膜HSP70蛋白及其mRNA水平的影響[J];中國腫瘤生物治療雜志;1998年01期

5 ;[J];;年期

相關會議論文 前1條

1 許睿;張姝;安保孝幸;楳田yP子;坂口e，

本文編號：2178378