摘要目的 了解1,25(OH)2D3对载脂蛋白E缺乏鼠(apoE-/-)主动脉内皮细胞(EC)增生、凋亡及内皮型一氧化氮合酶(eNOS)表达的影响.方法 apoE-/-鼠主动脉EC分离培养,MTT法观察1,25(OH)2D3影响apoE-/-鼠EC生长,TUNEL检测细胞凋亡,RT-PCR检测Bc1-2、Fas mRNA和eNOS mRNA表达.结果 细胞对照组与无水乙醇对照组EC增生率差异无统计学意义[(0.162±0.031)vs.(0.158±0.006),p＞0.05],1,25(OH)2D3在10-4、10-5、10-6、10-7、10-8 mol/L时EC增生率高于对照组(P＜0.01),但不同浓度1,25( OH)2D3作用组之间差异无统计学意义[分别为(0.189±0.013)、(0.285±0.011)、(0.296±0.026)、(0.284±0.017)、(0.233±0.010),P ＞0.05],选定10-6mol/L 1,25(OH)2D3为研究浓度,干预分离培养apoE-/-鼠主动脉EC.1,25( OH)2D310-6mol/L组、细胞对照组、无水乙醇对照组凋亡指数分别为(15.14±3.19)、(18.94 ±4.22)、(19.27±4.58),Bcl-2 mRNA分别为(0.78±0.16)、(0.46±0.21)、(0.42±0.17),Fas mRNA分别为(0.43±0.12)、(0.79±0.21)、(0.81±0.20),eNOS mRNA分别为(0.56±0.16)、(0.39±0.13)、(0.35±0.11).25(OH)2D3干预组EC凋亡指数、Fas mRNA、eNOS mRNA降低,Bcl-2 mRNA增高(P均＜0.01),细胞对照组与无水乙醇对照组比较差异无统计学意义(P＞0.05).相关分析发现在1,25(OH)2D3干预组,eNOS表达量与凋亡指数、Fas mRNA呈负相关(r=-0.676、-0.758),与Bcl-2表达呈正相关(r =0.762),差异有统计学意义(P均＜0.01).结论 1,25(OH)2D3刺激apoE-/-鼠主动脉EC增生、抑制主动脉EC凋亡,影响凋亡相关基因Bcl-2、Fas mRNA表达,上调eNOS mRNA表达.

abstractsObjective To study possible influences of 1,25 (OH)2D3 on endothelial cell proliferation,apoptosis and endothelial nitric oxide synthase (eNOS) expression of aorta in apolipoprotein E-deficient ( apoE -/- ) mice and to explore the relationship between vitamin D and atherosclerosis.Method Endothelial cell of aorta in apoE -/- mice were isolated and cultured,and the influence of 1,25 (OH)2 D3 on endothelial cell proliferation were observed by MTT,apoptosis of cells were quantitated by terminal deoxynucleotidyl transferase mediated dUTP nick end labelling,Bcl-2 mRNA,fas mRNA and eNOS mRNA was detected by reverse transcription-polymerase chain reaction.Result Endothelial cell proliferation rate of aorta did not significantly change in the two control groups (0.162 ±0.031 vs.0.158 ±0.006,P ＞0.05 ).Compared with control groups,1,25 (OH) 2 D3 stimulated endothelial cell proliferation of aorta ( P ＜0.05),but endothelial cell proliferation rate did not significantly change in different 1,25 (OH)2D3concentration groups [ 1,25( OH)2D3 concentration:10-4moL/L,10-5 moL/L,10-6 moL/L,10 -7 mol/L,10-8 mol/L,endothelial cell proliferation rate:0.189 ± 0.013 vs.0.285 ± 0.011 vs.0.296 ± 0.026 vs.0.284 ±0.017 vs.0.233 ± 0.010,P ＞ 0.05 ].1,25 ( OH )2D3 research concentration as chosen as 10 -6mol/L.In 1,25 (OH)2D3 10-6 mol/L group,the expression of Bcl-2,eNOS mRNA was significantly increased (0.78 t:0.16 vs.0.46 ± 0.21 vs.0.42 ± 0.17,0.56 ± 0.16 vs.0.39 ± 0.13 vs.0.35 ± 0.11,0.46 ± 0.2 vs.10.42 ± 0.17 vs.0.78 ± 0.16,0.79 ± 0.21 vs.0.81 ± 0.20 vs.0.43 ± 0.12 ),apoptotic index,Fas mRNA was significantly decreased ( 15.14 ±3.19 vs.18.94 ±4.22 vs.19.27 ±4.58,0.43 ±0.12 vs.0.79 ±0.21 vs.0.81 ± 0.20) ( P ＜ 0.05 ).The quantity of eNOS gene expression was inversely associated with apoptosis index and Fas mRNA,was positively associated with Bcl-2 mRNA (r =-0.676,-0.758,0.762,P ＜0.01).Conclusion 1,25(OH)2D3 stimulated endothelial cell proliferation,inhibited apoptosis and increased eNOS expression of aorta in apoE-/- mice.These results may deepen understanding of the pathogenesis of atherosclerosis.