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
发布时间：2018-08-19 11:42
【摘要】：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(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。
[Abstract]:Aim: to investigate the effects of oleuropein op and acrolein on the proliferation and apoptosis of human bronchial epithelial-like cells (human bronchial epithelial cells) under endoplasmic reticulum stress (endouplasmic reticulum stress). Methods the survival rate of HBE cells treated with ACR and op was determined by Hoechst33258 staining and apoptosis by flow cytometry. Glucose-regulated protein 78 GRP78 and CCAAT enhancer binding protein homologous eggs were detected by Western blot. The expression of C/EBP homologous protein chop was detected by Real-time PCR method. The expression of GRP78 and CHOP mRNA was detected by real-time PCR. Male Sprague-Dawley rats were treated by intraperitoneal injection of ACR and / or (olive leaf extract from olive leaves. The expression of ERS related protein GRP78 and CHOP were detected by Western blot in lung tissue of rats. GRP78 and CHOP mRNA were detected by real-time PCR. Results: in vitro, ACR could inhibit the proliferation and induce apoptosis of HBE cells, and the effect of ACR on inhibiting cell proliferation and promoting apoptosis was obviously inhibited after treatment with op. At the same time, ACR upregulated the expression of ERS related protein GRP78 and CHOP in HBE cells, combined with op could inhibit the expression of GRP78 and CHOP, and at mRNA level alone, ACR upregulated the mRNA expression of ERS related molecules GRP78 and CHOP. There was no significant change in mRNA levels of GRP78 and CHOP treated with op. The results of in vivo and in vitro cell experiments of Sprague-Dawley rats were consistent with those in vitro. ConclusionACR can inhibit the proliferation of HBE cells and induce the apoptosis of HBE cells. At the level of protein translation, op can antagonize the apoptosis induced by ACR by inhibiting the ERS pathway.
【作者单位】： 南京医科大学公共卫生学院营养与食品卫生学系;
【基金】：国家自然科学基金(81472977) 江苏高校优势学科建设工程资助项目
【分类号】：R151
，
本文编号：2191554
[Abstract]:Aim: to investigate the effects of oleuropein op and acrolein on the proliferation and apoptosis of human bronchial epithelial-like cells (human bronchial epithelial cells) under endoplasmic reticulum stress (endouplasmic reticulum stress). Methods the survival rate of HBE cells treated with ACR and op was determined by Hoechst33258 staining and apoptosis by flow cytometry. Glucose-regulated protein 78 GRP78 and CCAAT enhancer binding protein homologous eggs were detected by Western blot. The expression of C/EBP homologous protein chop was detected by Real-time PCR method. The expression of GRP78 and CHOP mRNA was detected by real-time PCR. Male Sprague-Dawley rats were treated by intraperitoneal injection of ACR and / or (olive leaf extract from olive leaves. The expression of ERS related protein GRP78 and CHOP were detected by Western blot in lung tissue of rats. GRP78 and CHOP mRNA were detected by real-time PCR. Results: in vitro, ACR could inhibit the proliferation and induce apoptosis of HBE cells, and the effect of ACR on inhibiting cell proliferation and promoting apoptosis was obviously inhibited after treatment with op. At the same time, ACR upregulated the expression of ERS related protein GRP78 and CHOP in HBE cells, combined with op could inhibit the expression of GRP78 and CHOP, and at mRNA level alone, ACR upregulated the mRNA expression of ERS related molecules GRP78 and CHOP. There was no significant change in mRNA levels of GRP78 and CHOP treated with op. The results of in vivo and in vitro cell experiments of Sprague-Dawley rats were consistent with those in vitro. ConclusionACR can inhibit the proliferation of HBE cells and induce the apoptosis of HBE cells. At the level of protein translation, op can antagonize the apoptosis induced by ACR by inhibiting the ERS pathway.
【作者单位】： 南京医科大学公共卫生学院营养与食品卫生学系;
【基金】：国家自然科学基金(81472977) 江苏高校优势学科建设工程资助项目
【分类号】：R151
，
本文编号：2191554
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