[關(guān)鍵詞]
[摘要]
目的 探討注射用益氣復(fù)脈(凍干,YQFM)對(duì)阿霉素(doxorubicin,DOX)誘導(dǎo)H9c2(2-1)心肌細(xì)胞毒性的保護(hù)作用。方法 H9c2(2-1)心肌細(xì)胞隨機(jī)分為對(duì)照組,模型組(終濃度為0.3 μmol/L的DOX處理細(xì)胞48 h),YQFM低、中、高劑量(125、625、3 125 μg/mL)組(提前2 h加入藥物預(yù)處理,然后加入終濃度為0.3 μmol/L的DOX處理48 h),采用CCK-8法檢測(cè)細(xì)胞活力;使用乳酸脫氫酶(LDH)和ATP試劑盒檢測(cè)細(xì)胞LDH和ATP水平;應(yīng)用Hoechst 33258染色法檢測(cè)細(xì)胞凋亡;JC-1法檢測(cè)細(xì)胞線粒體膜電位;Western blotting法檢測(cè)caspase-3蛋白的表達(dá)水平。結(jié)果 與模型組比較,YQFM中、高劑量組顯著增加細(xì)胞存活率(P<0.05、0.01),低、高劑量組明顯改善細(xì)胞凋亡;低、中、高劑量組LDH活性顯著降低(P<0.05、0.01),ATP含量顯著增加(P<0.05、0.01),線粒體膜電位明顯恢復(fù)。結(jié)論 YQFM抑制DOX誘導(dǎo)H9c2(2-1)的細(xì)胞毒性,其作用機(jī)制可能與抑制線粒體凋亡信號(hào)通路的激活有關(guān)。
[Key word]
[Abstract]
Objective To investigate the protective effect of Yiqi Fumai Lyophilized Injection (YQFM) against Doxorubicin (Dox)-induced cardiotoxicity in H9c2 (2-1). Methods H9c2 (2-1) myocardial cells were randomly divided into control group, model group (cells treated with DOX at the final concentration of 0.3 μmol/L for 48 h), YQFM low, medium and high dose (125, 625, and 3 125 g/mL) group (pretreatment with YQFM for 2 h, and then adding DOX of a final concentration of 0.3 μmol/L). Cell viability was detected with CCK-8 assay. Cells LDH and ATP levels were examined using LDH and ATP kits. Cell death was measured by Hoechst 33258 stain assay. The mitochondrial membrane potential was detected by JC-1. Caspase-3 protein expression was evaluated by Western blotting. Results Compared with model group, YQFM medium and high dose group significantly increased the cell survival rate (P < 0.05 and 0.01), low and high dose group significantly improved cell apoptosis; LDH activity of low, medium and high dose group was significantly decreased (P < 0.05 and 0.01), ATP content increased significantly (P < 0.05 and 0.01), and the mitochondrial membrane potential was restored. Conclusion YQFM inhibited DOX-induced cardiotoxicity in H9c2 (2-1), the possible mechanisms may be related to the inhibition of activation of mitochondrial apoptotic signaling pathway.
[中圖分類號(hào)]
[基金項(xiàng)目]
天津市中藥注射劑關(guān)鍵技術(shù)校企協(xié)同創(chuàng)新實(shí)驗(yàn)室建設(shè)基金(17PTSYJC00090)