橄榄苦苷抑制丙烯醛诱导的HBE细胞内质网应激的机制研究.docxVIP

橄榄苦昔抑制丙烯醛诱导的HBE细胞内 质网应激的机制研究 徐朝琪许玉宇姜盼冯晴 南京医科大学公共卫生学院营养与食品卫生学系 摘要: 目的:探讨橄榄苦昔(oleuropein, OP)和丙烯醛(acrolein, ACR)对人支气管 上皮样细胞(human bronchial epithelial cells, HBE)内质网应激 (endouplasmic reticulum stress, ERS)相关的增殖与凋亡影响的可能机制。 方法：0P与ACR联合处理HBE细胞，MTT比色法检测细胞存活率,Hoechst33258 染色法和流式细胞检测细胞凋亡，Western blol检测ERS相关蛋白葡萄糖调节 蛋白 78 (glucose-regulated protein 78, GRP78)和 CCAAT 增强子结合蛋白同 源蛋口 (C/EBP homologous protein, CHOP)的表达，Real-time PCR 法检测 GRP78和CHOP mRNA表达;雄性Sprague-Dawley大鼠经ACR腹腔注射和(或)橄 榄叶提取物(olive leaf extract, OLE)灌胃处理，取大鼠肺组织‘Western blot法检测ERS相关蛋白GRP78和CHOP的表达,Real-time PCR法检测GRP78 和CHOP mRNA表达。结果:体外实验证实ACR可以抑制IIBE细胞的增殖，诱导其 凋亡;联合0P处理后，ACR抑制细胞增殖、促进细胞凋亡的作用明显受到抑制； 同时，ACR上调HBE细胞中ERS相关蛋白GRP78和CHOP的表达，联合0P可抑制 GRP78和CHOP的表达;在mRNA水平上，ACR单独处理上调了 ERS相关分子GRP78 和CHOP的mRNA表达水平，而联合0P处理，GRP78和CHOP的mRNA水平未见明 显变化;Sprague-Dawley大鼠体内实验结果与体外细胞实验基本一致。结论:ACR 抑制HBE细胞增殖，促发ERS,进而诱导HBE细胞凋亡。在蛋白质翻译水平上，0P 可以通过抑制ERS途径，拮抗ACR诱导的细胞凋亡效应。 关键词： 橄榄苦晋;丙烯醛;IIBE细胞;内质网应激; 作者简介：冯晴，E-mail :qingfcng@njmu. cdu. cn 收稿日期:2017-02-24 基金：国家自然科学基金 Mechanism study of oleuropein inhibiting endoplasmic reticulum stress induced by acrolein in HBE cells Xu Chaoqi Xu Yuyu Jiang Pan Feng Qing Department of Nutrition and Food Hygiene, School of Public Health, NJMU; Abstract： Objective:To study the effect of oleuropein in combination of acrolein on proliferation and apoptosis related to endoplasmic reticulum stress (ERS) in human bronchial epithelial cells. Methods:HBE cells were treated with oleuropein and acrolein. MTT assay was performed to detect cel 1 viability. Plate clone formation assay was used to get colony forming efficiency. Hocchst33258 staining and flow cytomctrie assay were applied to investigate the apoptosis of cells.The levels of ERS biomarkers including glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) were measured by Western blot. The mRNA expressions of GRP78 and CHOP were dnalyzed by real-time PCR- Twelve male Sprague-Dawley rats were treated with acrolein alone through intrapcritone