【摘要】：目的·研究低糖后復(fù)糖對H9c2大鼠心肌細(xì)胞自噬活性的影響。方法·將H9c2細(xì)胞分為低糖組(LG)、復(fù)糖組(RG)、自噬抑制組[RG+氯喹(CQ)]和空白對照組(Con)。LG組用低糖培養(yǎng)基培養(yǎng)2 h;RG組用低糖培養(yǎng)基培養(yǎng)2 h后,用高糖培養(yǎng)基繼續(xù)培養(yǎng)2 h;RG+CQ組在給予低糖和高糖處理前1 h加入CQ;Con組始終用高糖培養(yǎng)基培養(yǎng)。采用MTT法檢測H9c2細(xì)胞增殖能力,LDH活性檢測試劑盒檢測細(xì)胞毒性,Western blotting檢測p-m TOR、m TOR、p-AMPK、AMPK、LC3、beclin-1的表達(dá)量。結(jié)果·與LG組相比,RG組細(xì)胞增殖能力降低,LDH活性升高(均P0.05);與RG組相比,RG+CQ組細(xì)胞增殖能力進一步降低,LDH活性進一步升高(均P0.05)。Western blotting結(jié)果顯示,與LG組相比,RG組p-AMPK、LC3Ⅱ/Ⅰ和beclin-1表達(dá)量增加,p-m TOR表達(dá)量降低(均P0.05);RG+CQ組較RG組LC3Ⅱ/Ⅰ和beclin-1表達(dá)進一步增加(均P0.05)。結(jié)論·H9c2細(xì)胞經(jīng)低糖后復(fù)糖處理后自噬活性增強,這可能與激活A(yù)MPK/m TOR途徑有關(guān),且此時自噬活性增強對于心肌細(xì)胞具有保護作用。
[Abstract]:Objective to study the effect of hypoglycemic compound on autophagy of cardiac myocytes in H9c2 rats. Methods H9c2 cells were divided into low glucose group, (LG), complex group, (RG), autophagy inhibition group [RG chloroquine (CQ)] and blank control group (Con). LG group, cultured in low glucose medium for 2 hours. The RG group was cultured in low glucose medium for 2 h, and the high glucose medium was used in the high glucose medium for 2 h. The RG CQ group was added to the CQ;Con group 1 h before being treated with low glucose and high glucose, and the high glucose medium was used all the time. The proliferation ability of H9c2 cells was detected by MTT assay, and the expression of pAm TOR,m TOR,p-AMPK,LC3,beclin-1 was detected by cytotoxic, Western blotting assay with LDH activity assay kit. Results compared with LG group, the proliferation ability of RG group decreased and the activity of LDH increased (P0.05). Compared with RG group, the proliferation ability of, RG CQ group was further decreased and the activity of LDH was further increased (P0.05). Western blotting). Compared with LG group, the expression of LC3 鈪，

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