探索发状分裂相关增强子1（HES-1）差异表达对胆固醇刺激下血管内皮细胞的影响。

将Ea.hy926细胞分为4组，即NC组（空白对照）、CH组（单纯胆固醇刺激）、GO组（HES-1过表达）和GD组（HES-1低表达）。将HES-1片段克隆到载体中构建pCDNA3.1-3×Flag-HES-1β过表达载体，转染细胞后构建HES-1过表达细胞，用退火寡糖和消化的CRISPR载体构建HES-1基因敲除细胞，Western blot检测细胞中HES-1表达水平。选取不同浓度胆固醇（50、100和200 mmol/L）刺激细胞不同的时间（12、24和48 h），MTT法检测细胞存活率并选定适宜造模条件。按照确定的胆固醇作用浓度和时间刺激CH、GO、GD组细胞，采用流式细胞术检测细胞凋亡水平。实时荧光定量PCR法检测凋亡相关基因TNF-α、Bax、NF-κB和cyclin D1的相对表达水平。

在100 mmol/L胆固醇刺激24 h下，与NC组比较，CH组和GO组的细胞存活率显著降低，差异均有统计学意义（P<0.05），GD组的细胞存活率降低，但差异无统计学意义（P>0.05）；与CH组比较，GO组细胞存活率降低，GD组细胞存活率升高，差异均有统计学意义（P<0.05），故选用100 mmol/L胆固醇刺激24 h作为造模条件。与NC组（0.03%±0.01%）比较，CH组、GO组和GD组凋亡细胞百分比（36.03%±3.21%、56.73%±5.48%、20.03%±2.03%）均显著增加，其中GO组增加最显著，差异均有统计学意义（P<0.05）。与CH组相比，GO组TNF-α、Bax表达升高，cyclin D1和Bcl-2表达降低；GD组cyclin D1和Bcl-2表达升高，Bax表达降低；差异均有统计学意义（P<0.05）。

在胆固醇刺激下，HES-1过表达可激活凋亡相关基因表达，促进血管内皮细胞凋亡。

To explore the effect of differential expression of hairy and enhancer of split homolog-1 (HES-1) on cholesterol-stimulated vascular endothelial cells.

Ea.hy926 cells were divided into four groups, including NC group without cholesterol stimulation, CH group with cholesterol stimulation alone, GO group with cholesterol stimulation combined with HES-1 overexpression, and GD group with cholesterol stimulation combined with HES-1 low expression. The HES-1 fragment was cloned into vector to construct pCDNA3.1-3×Flag-HES-1β overexpression vector, and HES-1 overexpression cells were constructed after transfection of cells; HES-1 knockdown cells were constructed with annealed oligosaccharide and digested CRISPR vector. HES-1 expression level was detected by Western blot. The cells were stimulated with different concentrations of cholesterol (50, 100 and 200 mmol/L) for different times (12, 24 and 48 h), and the MTT method was used to detect the cell viability and select the appropriate modeling conditions. The cells of CH, GO and GD groups were stimulated according to the determined cholesterol concentration and time, and the apoptosis levels were detected by flow cytometry. The relative expression levels of apoptosis-related genes TNF-α, Bax, NF-κB and cyclin D1 were detected by real-time PCR and flow cytometry.

Under 100 mmol/L cholesterol stimulation for 24 h, compared with the NC group, the cell survival rates of CH and GO groups were significantly lower (P<0.05), and the cell survival rates of GD group were not statistically significant (P>0.05). Therefore, 100 mmol/L cholesterol stimulation for 24 h was selected as the modeling condition. Compared with the NC group (0.03%±0.01%), the percentage of apoptotic cells in the CH group, GO group, and GD group (36.03%±3.21%, 56.73%±5.48%, 20.03%±2.03%) significantly increased, with the GO group showing the most significant increase, with statistically significant differences (P<0.05). Compared with the CH group, TNF-α and Bax were elevated and cyclin D1 and Bcl-2 expression were decreased in the GO group; cyclin D1 and Bcl-2 expression was elevated and Bax was decreased in the GD group, with statistically significant differences (P<0.05).

Under cholesterol stimulation, HES-1 overexpression can activate apoptosis-related gene expression and promote vascular endothelial cell apoptosis.