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目的 :研究 3β ,5α ,6 β 胆甾烷三醇 (Triol)和2 5 羟胆固醇 (2 5OH)对VSMC增殖的影响 ,探讨氧化甾醇在动脉粥样硬化 (AS)发病中的作用和意义。方法 :培养和鉴定新西兰白兔主动脉VSMC ;以含不同浓度的Triol和 2 5OH的培养液作用于VSMC为实验组 ,不含氧化甾醇或含胆固醇的培养液作为对照组。VSMC增殖及其代谢活性以WST 1测定 ,细胞凋亡以光镜、透射电镜、脱氧核苷酸末端转移酶介导脱氧尿苷三磷酸缺口末端标记 (TUNEL)判断。结果 :在1× 10 - 9～ 1× 10 - 7mol·L- 1范围的Triol和 2 5OH对VSMC的WST 1代谢活性无明显变化 (P >0 .0 5 ) ,而≥ 1× 10 - 6 mol·L- 1的氧化甾醇则使其代谢活性显著下降 (P <0 .0 1) ;≥ 2 0× 10 - 6 mol·L- 1的Triol和2 5OH诱导VSMC胞体皱缩变小、染色质在核周边浓集、核碎裂、空泡和凋亡小体形成等超微结构变化 ;TUNEL呈阳性反应。结论 :小剂量氧化甾醇无促进VSMC增殖而较大剂量氧化甾醇可诱导VSMC凋亡。
Objective: To investigate the effect of 3β, 5α, 6β triol and 25-hydroxycholesterol (2 5OH) on VSMC proliferation and to explore the role and significance of oxidized sterol in the pathogenesis of atherosclerosis (AS). METHODS: New Zealand white rabbits were aortic VSMCs cultured and identified. Serums containing different concentrations of Triol and 25OH were used as the experimental group and without oxysterol or cholesterol-containing culture medium as the control group. The proliferation and the metabolic activity of VSMC were measured by WST 1. The apoptotic cells were identified by light microscopy, transmission electron microscopy and deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL). Results: There was no significant change in WST 1 metabolic activity of VSMCs between 1 × 10 - 9 and 1 × 10 - 7 mol·L - 1 Triol and 2 5OH (P> 0.05), while ≥ 1 × 10 - 6 (P <0.01). Triol and 2 5OH of ≥20 × 10-6 mol·L-1 induced a decrease in the shrinkage of VSMC somatic cells, staining The ultrastructural changes such as the concentration around the nucleus, the nuclear fragmentation, the vacuolization and the formation of apoptotic bodies; TUNEL positive reaction. Conclusion: Small doses of oxidized sterol did not promote the proliferation of VSMC, but larger doses of oxidized sterol induced VSMC apoptosis.