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目的观察二氮嗪预处理对大鼠缺氧-复氧(H-R)心肌微血管内皮细胞(MMECs)Fractalkine(FKN)和p53 mRNA表达的影响。方法培养SD大鼠MMECs,分为四组:正常对照组(N组)、H-R组、二氮嗪组(DZ组)、二氮嗪+mitoKATP特异性阻断剂5-羟葵酸预处理组(DZ+5-HD组)。DZ组和DZ+5-HD组分别加入二氮嗪100μmol/L、二氮嗪100μmol/L+5-羟葵酸100μmol/L预处理2 h,然后和H-R组同样进行H-R 2 h。观察凋亡细胞形态,测定细胞凋亡率和活力、RT-PCR检测FKN和p53 mRNA转录水平。结果与N组比较,H-R组细胞凋亡率显著升高,细胞增殖率显著下降,p53 mRNA和FKN mRNA表达显著增强(P<0.01);与H-R组比较,DZ组细胞凋亡率显著降低,细胞增殖率显著增强(P<0.05),FKN mRNA和p53 mRNA表达显著降低(P<0.01);DZ+5-HD组取消了DZ组的作用。结论二氮嗪预处理通过抑制细胞凋亡,促使细胞增殖,下调FKN mRNA和p53 mRNA表达而实现对H-R MMECs损伤的保护。
Objective To observe the effect of diazoxide preconditioning on Fractalkine (FKN) and p53 mRNA expression in hypoxia-reoxygenation (H-R) myocardium microvascular endothelial cells (MMECs). Methods SD rat MMECs were cultured and divided into four groups: normal control group (N group), HR group, diazoxide group (DZ group), diazoxide + mitoKATP specific antagonist 5-hydroxalic acid pretreatment group (DZ + 5-HD group). DZ group and DZ + 5-HD group were added diazoxide 100μmol / L, diazoxide 100μmol / L + 5-hydroxy acid 100μmol / L pretreatment 2 h, and then H-R group H-R 2 h. The morphology of apoptotic cells was observed, the apoptosis rate and viability were measured, and the transcriptional levels of FKN and p53 mRNA were detected by RT-PCR. Results Compared with the N group, the apoptotic rate of HR group was significantly increased, the cell proliferation rate was significantly decreased, and the expression of p53 mRNA and FKN mRNA was significantly increased (P <0.01); Compared with HR group, the apoptosis rate of DZ group was significantly decreased, (P <0.05). The expression of FKN mRNA and p53 mRNA was significantly decreased (P <0.01). The DZ group was abolished in DZ + 5-HD group. Conclusion Diazoxide preconditioning can protect H-R MMECs by inhibiting cell apoptosis, promoting cell proliferation and down-regulating FKN mRNA and p53 mRNA expression.