【摘要】：目的:探討成纖維細(xì)胞生長(zhǎng)因子21(FGF21)在內(nèi)質(zhì)網(wǎng)應(yīng)激(ERS)誘導(dǎo)心肌細(xì)胞凋亡中的保護(hù)作用及其作用機(jī)制。方法:以pc DNA4為基因載體,構(gòu)建pc DNA4-FGF21質(zhì)粒并轉(zhuǎn)染H9c2大鼠心肌細(xì)胞48h,加入衣霉素(TM)10μM處理24h構(gòu)建內(nèi)質(zhì)網(wǎng)應(yīng)激模型,實(shí)驗(yàn)分為4個(gè)組:對(duì)照組、衣霉素處理組、pcDNA4-FGF21+衣霉素組、pcDNA4+衣霉素組。利用蛋白免疫印跡(Western blot)方法檢測(cè)FGF21蛋白及蛋白激酶R樣ER激酶(PERK)和c-Jun氨基末端激酶(JNK)介導(dǎo)的凋亡通路相關(guān)蛋白的表達(dá)。利用CCK8檢測(cè)細(xì)胞存活率和TUNEL檢測(cè)細(xì)胞凋亡率。結(jié)果:成功構(gòu)建pcDNA4-FGF21質(zhì)粒并在H9c2細(xì)胞中過(guò)表達(dá)。與對(duì)照組相比,衣霉素處理組和pcDNA4+衣霉素組明顯上調(diào)H9c2細(xì)胞內(nèi)源性FGF21的表達(dá)(P0.01),以及增加PERK和JNK介導(dǎo)的凋亡通路相關(guān)蛋白的表達(dá)(P0.05~0.01),減少細(xì)胞存活率和提高細(xì)胞凋亡水平(P0.05~0.01)。與衣霉素處理組和pcDNA4+衣霉素組相比,在pcDNA4-FGF21+衣霉素組明顯降低PERK和JNK介導(dǎo)的凋亡通路相關(guān)蛋白的表達(dá),增加細(xì)胞存活率,降低細(xì)胞凋亡水平(P0.05~0.01)。結(jié)論:FGF21過(guò)表達(dá)可以減輕內(nèi)質(zhì)網(wǎng)應(yīng)激誘導(dǎo)心肌細(xì)胞的凋亡,其機(jī)制可能部分與抑制內(nèi)質(zhì)網(wǎng)應(yīng)激中PERK和JNK介導(dǎo)促凋亡通路的信號(hào)傳導(dǎo)有關(guān)。
[Abstract]:Aim: to investigate the protective effect of fibroblast growth factor 21 (FGF21) on cardiac myocyte apoptosis induced by endoplasmic reticulum stress (ERS) and its mechanism. Methods: pc DNA4 was used as gene vector, pc DNA4-FGF21 plasmid was constructed and transfected into H9c2 rat cardiomyocytes for 48 h. Endoplasmic reticulum stress model was established by adding 10 渭 M (TM) for 24 hours. The model was divided into four groups: control group and Itetracycline treated group. PcDNA4-FGF21 group and pcDNA4 group. Western blot (Western blot) was used to detect the expression of FGF21 protein, protein kinase R-like ER kinase (PERK) and c-Jun amino-terminal kinase (JNK) mediated apoptosis pathway. Cell survival rate was detected by CCK8 and apoptosis rate was detected by TUNEL. Results: pcDNA4-FGF21 plasmid was successfully constructed and overexpressed in H9c2 cells. Compared with the control group, the expression of endogenous FGF21 in H9c2 cells and the expression of PERK and JNK mediated apoptosis-related proteins in H9c2 cells were upregulated in the chlortetracycline and pcDNA4 groups (P0.05 / 0. 01). The cell survival rate was reduced and the apoptosis level was increased (P0.05N0.01). Compared with the pcDNA4-FGF21 group and the pcDNA4 group, the expression of apoptosis-related proteins mediated by PERK and JNK was significantly decreased, the cell survival rate was increased, and the apoptosis level was decreased in the pcDNA4-FGF21 group (P0.05 / 0. 01). Conclusion: overexpression of FGF21 can attenuate the apoptosis induced by endoplasmic reticulum stress, and its mechanism may be related to the inhibition of signal transduction mediated by PERK and JNK in endoplasmic reticulum stress.
【作者單位】： 青島大學(xué)醫(yī)學(xué)院附屬煙臺(tái)毓璜頂醫(yī)院心內(nèi)科;青島大學(xué)醫(yī)學(xué)院附屬煙臺(tái)毓璜頂醫(yī)院生物芯片;青島大學(xué)醫(yī)學(xué)院附屬煙臺(tái)毓璜頂醫(yī)院中心實(shí)驗(yàn)室;
【分類(lèi)號(hào)】：R54

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