【摘要】：目的:探討橄欖苦苷(oleuropein,OP)和丙烯醛(acrolein,ACR)對(duì)人支氣管上皮樣細(xì)胞(human bronchial epithelial cells,HBE)內(nèi)質(zhì)網(wǎng)應(yīng)激(endouplasmic reticulum stress,ERS)相關(guān)的增殖與凋亡影響的可能機(jī)制。方法 :OP與ACR聯(lián)合處理HBE細(xì)胞,MTT比色法檢測(cè)細(xì)胞存活率,Hoechst33258染色法和流式細(xì)胞檢測(cè)細(xì)胞凋亡,Western blot檢測(cè)ERS相關(guān)蛋白葡萄糖調(diào)節(jié)蛋白78(glucose-regulated protein 78,GRP78)和CCAAT增強(qiáng)子結(jié)合蛋白同源蛋白(C/EBP homologous protein,CHOP)的表達(dá),Real-time PCR法檢測(cè)GRP78和CHOP mRNA表達(dá);雄性Sprague-Dawley大鼠經(jīng)ACR腹腔注射和(或)橄欖葉提取物(olive leaf extract,OLE)灌胃處理,取大鼠肺組織,Western blot法檢測(cè)ERS相關(guān)蛋白GRP78和CHOP的表達(dá),Real-time PCR法檢測(cè)GRP78和CHOP mRNA表達(dá)。結(jié)果:體外實(shí)驗(yàn)證實(shí)ACR可以抑制HBE細(xì)胞的增殖,誘導(dǎo)其凋亡;聯(lián)合OP處理后,ACR抑制細(xì)胞增殖、促進(jìn)細(xì)胞凋亡的作用明顯受到抑制;同時(shí),ACR上調(diào)HBE細(xì)胞中ERS相關(guān)蛋白GRP78和CHOP的表達(dá),聯(lián)合OP可抑制GRP78和CHOP的表達(dá);在mRNA水平上,ACR單獨(dú)處理上調(diào)了ERS相關(guān)分子GRP78和CHOP的mRNA表達(dá)水平,而聯(lián)合OP處理,GRP78和CHOP的mRNA水平未見明顯變化;Sprague-Dawley大鼠體內(nèi)實(shí)驗(yàn)結(jié)果與體外細(xì)胞實(shí)驗(yàn)基本一致。結(jié)論:ACR抑制HBE細(xì)胞增殖,促發(fā)ERS,進(jìn)而誘導(dǎo)HBE細(xì)胞凋亡。在蛋白質(zhì)翻譯水平上,OP可以通過抑制ERS途徑,拮抗ACR誘導(dǎo)的細(xì)胞凋亡效應(yīng)。
[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.
【作者單位】： 南京醫(yī)科大學(xué)公共衛(wèi)生學(xué)院營(yíng)養(yǎng)與食品衛(wèi)生學(xué)系;
【基金】：國(guó)家自然科學(xué)基金(81472977) 江蘇高校優(yōu)勢(shì)學(xué)科建設(shè)工程資助項(xiàng)目
【分類號(hào)】：R151
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本文編號(hào)：2191554