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目的:探讨橄榄苦苷(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:OP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 blot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白(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(o-live 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HBE细胞的增殖,诱导其凋亡;联合OP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OP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OP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,OP可以通过抑制ERS途径,拮抗ACR诱导的细胞凋亡效应.“,”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 cell viability.Plate clone formation assay was used to get colony forming efficiency.Hoechst 33258 staining and flow cytometric 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 analyzed by real-time PCR.Twelve male Sprague-Dawley rats were treated with acrolein alone through intraperitoneal injection,or olive leaf extract in combination through gavage.Lung tissues were obtained from the rats for detecting the expressions of GRP78 and CHOP.Results:HBE cell proliferation was inhibited by acrolein in a dose dependent manner.Cell proliferation increased significantly with the combination of oleuropein.Oleuropein combined with acrolein also inhibited acrolein-induced apoptosis in HBE cells.Western blot demonstrated that ERS biomarkers such as GRP78 and CHOP were induced by acrolein combined with oleuropein,acrolein-induced ERS was alleviated.However,oleuropein had no effects on the GRP78 and CHOP mRNA expression.The in vivo results of Sprague-Dawley rats were in agreement with those in vitro.Conclusion:Acrolein could trigger ERS in HBE cells and lead to cell apoptosis.Nevertheless,oleuropein in combination with acrolein could inhibit the acrolein-induced apoptosis.The potential mechanism may be related with the alleviation of acrolein-induced ERS.In protein translational level,oleuropein can inhibit apoptosis induced by acrolein via ERS pathway.