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目的 观察金属硫蛋白 (metallothionein ,MT)对同型半胱氨酸损伤血管内皮细胞的拮抗作用。方法 人血管内皮细胞培养 ;自动生化分析仪测乳酸脱氢酶 (LDH)漏出量 ;10 9Cd 血红素饱和法测细胞MT含量 ;台盼蓝排除法计算细胞存活率。结果 同型半胱氨酰 (Hcy) (0 1～ 1 0mmol·L-1)呈浓度依赖性地增加细胞LDH漏出和降低细胞存活率。外源给予MT 5× 10 -8μmol·L-1可呈剂量依赖性的抑制Hcy引起的血管内皮细胞损伤 ,5× 10 -8mol·L-1MT与1 0mmol·L-1Hcy共同孵育组较单纯Hcy孵育组血管内皮细胞LDH漏出降低 2 7 1% (P <0 0 5 ) ,细胞存活增加 6 %(P <0 0 5 )。凝血酶预孵育可诱导培养的血管内皮细胞MT生成 ,其含量较对照组增加 32 5 % (P <0 0 1)。凝血酶诱导内源性MT生成亦明显抑制Hcy引起的血管内皮细胞损伤 ,10 0 0U·L-1凝血酶与Hcy共同孵育组较单纯Hcy孵育组血管内皮细胞LDH漏出降低 46 % (P <0 0 1) ;细胞存活率升高 8% (P <0 0 5 )。结论 无论外源给予MT或内源性诱导MT生成均可降低Hcy引起的血管内皮细胞损伤作用
Objective To observe the antagonism of metallothionein (MT) on homocysteine-injured vascular endothelial cells. Methods Human vascular endothelial cells were cultured. The leakage of lactate dehydrogenase (LDH) was measured by automatic biochemical analyzer. The MT content of cells was measured by 10 9Cd heme saturation method. Cell viability was calculated by trypan blue exclusion method. Results Hcy (0 1 ~ 10 mmol·L-1) increased LDH leakage and decreased cell viability in a concentration-dependent manner. Exogenous administration of MT 5 × 10-8μmol·L-1 could inhibit Hcy-induced vascular endothelial cell injury in a dose-dependent manner. Co-incubation with 5 × 10 -8 mol·L-1MT and 10 mmol·L-1Hcy In the incubation group, LDH leakage decreased by 27.1% (P <0.05) and cell survival increased by 6% (P <0.05). Thrombin preincubation could induce MT formation of cultured vascular endothelial cells, which was increased by 325% (P <0.01) compared with the control group. Thrombin-induced endogenous MT production also significantly inhibited vascular endothelial cell injury induced by Hcy. Compared with Hcy-only incubation group, LDH leakage decreased by 46% (P <0) in HUVECs incubated with 10 000 U · L-1 thrombin and Hcy 0 1). Cell viability increased by 8% (P <0 05). Conclusion Exogenous MT or endogenous induced MT can reduce the injury of vascular endothelial cells induced by Hcy