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目的:探讨同型半胱氨酸(homocysteine,Hcy)对大鼠血管平滑肌细胞(vascular smooth muscle cells,v SMCs)增殖效应的影响及其与Ⅰ、Ⅲ、Ⅳ、Ⅵ型胶原产生间的关联。方法:用不同浓度(0.025~5.000 mmol/L)的Hcy刺激v SMCs,在不同时间点采用四甲基偶氮唑盐(MTT)法检测Hcy对平滑肌细胞活性的影响;5溴-2脱氧尿苷法检测Hcy对v SMCs DNA合成的作用,流式细胞仪观察细胞周期的变化;采用酶联免疫吸附试验测定胶原的含量。结果 :0.025~0.050 mmol/L的Hcy刺激12、48 h,v SMCs增殖明显增加。流式细胞仪分析发现,0.025~0.050 mmol/L的Hcy组其增殖细胞S期的百分比较对照组及其他浓度组明显增加。与对照组比较,细胞分泌的Ⅰ型和Ⅳ型胶原随着Hcy浓度的升高呈剂量依赖性增高。Ⅲ型胶原有轻度升高趋势,但差异尚无统计学意义。1~5 mmol/L的Hcy可减少Ⅵ型胶原的分泌,并呈剂量依赖性。结论:0.025~0.050 mmol/L的Hcy能促进v SMC增殖,且Hcy可能通过改变纤维斑块中的胶原类型构成导致斑块的不稳定性和易损性,最终加速了粥样斑块的发生和发展。
Objective: To investigate the effect of homocysteine (Hcy) on the proliferation of rat vascular smooth muscle cells (v SMCs) and its correlation with type Ⅰ, Ⅲ, Ⅳ and type Ⅵ collagen production. Methods: VSMCs were stimulated with Hcy at different concentrations (0.025-5.000 mmol / L), the effects of Hcy on the activity of SMCs were detected by MTT assay at different time points, and 5-bromo-2-deoxyuria Glycosides were used to detect the effect of Hcy on the DNA synthesis of v SMCs. The changes of cell cycle were observed by flow cytometry. The content of collagen was measured by enzyme linked immunosorbent assay (ELISA). Results: Hcy at 0.025-0.050 mmol / L stimulated for 12,48 h and v SMCs proliferated significantly. Flow cytometry analysis found that 0.025 ~ 0.050 mmol / L of Hcy group, the percentage of proliferating cells S phase was significantly increased compared with the control group and other concentrations. Compared with the control group, the type Ⅰ and type Ⅳ collagen secreted by cells increased in a dose-dependent manner with the increase of Hcy concentration. Type Ⅲ collagen increased slightly, but the difference was not statistically significant. 1 ~ 5 mmol / L Hcy can reduce the type Ⅵ collagen secretion in a dose-dependent manner. CONCLUSION: Hcy between 0.025 and 0.050 mmol / L can promote v SMC proliferation, and Hcy may accelerate the plaque instability and vulnerability by altering the collagen type in the fibrous plaque And development.