槲皮素对凝血酶诱导大鼠血管平滑肌细胞增殖和迁移的影响及其机制

Effects and related mechanism of quercetin on thrombin-induced proliferation and migration of ratvascular smooth muscle cells

目的探讨槲皮素对凝血酶诱导大鼠血管平滑肌细胞（VSMC）增殖和迁移的影响及其机制。方法取第3～5代体外培养的大鼠胸主动脉VSMC，分为对照组（在DMEM培养基中加入磷酸盐缓冲液）、凝血酶组（在DMEM培养基中加入1 U/ml凝血酶）和槲皮素组（在DMEM培养基中先加入1 U/ml凝血酶刺激后，再加入100 μmol/L槲皮素）。采用CCK－8法检测细胞的增殖能力；Transwell小室法检测细胞的迁移能力；Western blot法检测磷酸化细胞外信号调节激酶1/2（ERK 1/2）和磷酸化p38蛋白的表达水平。结果（1）CCK－8法检测显示，凝血酶组的吸光度值高于对照组（2.59±0.16比1.97±0.18，P〈0.01）；槲皮素组的吸光度值（2.13±0.19）低于凝血酶组（P〈0.01）；槲皮素组与对照组的吸光度值差异无统计学意义（P〉0.05）。（2）Transwell小室法检测显示，凝血酶组VSMC的迁移细胞数多于对照组[（1 337±162）个比（99±26）个，P〈0.01]；槲皮素组VSMC的迁移细胞数为（926±111）个，少于凝血酶组（P〈0.05）；槲皮素组VSMC的迁移细胞数多于对照组（P〈0.01）。（3）凝血酶组的磷酸化ERK 1/2和磷酸化p38蛋白表达水平均高于对照组（P均〈0.05）；槲皮素组的磷酸化ERK 1/2和磷酸化p38蛋白表达水平均低于凝血酶组（P均〈0.05）；槲皮素组与对照组的磷酸化ERK 1/2和磷酸化p38蛋白表达水平差异无统计学意义（P均〉0.05）。结论槲皮素可抑制凝血酶诱导的大鼠VSMC增殖和迁移，其机制可能与抑制ERK 1/2和p38磷酸化有关。

Objective To investigate the effects and related mechanism of quercetin on thrombin- induced proliferation and migration of rat vascular smooth muscle cells （VSMCs）. Methods Third to fifth generation VSMCs were divided into three groups, including control group （ with PBS on the base of medium DMEM）, thrombin group （with 1 U/ml thrombin on the base of medium DMEM） and quercetin group （treated with 100 μmol/L quercetin before 1 U/ml thrombin on the base of medium DMEM）. The proliferation and migration eapacitities were tested by CCK-8 kit and transwell chamber, respectively. The protein level of phospho-extraeellular signal regulated kinase 1/2 and phosphor-p38 were measured by Western blot. Results （ 1 ） According to the CCK-8 results, optical density value was significantly higher in thrombin group than that of control group （ 2. 59± 0. 16 vs. 1.97 ± 0. 18, P 〈 0. 01 ）, which could be significantly attenuated by pretreatment with quercetin （2. 13 ± 0. 19, P 〈 0. 01 ）, and there was no significant difference between quereetin group and control group（ P 〉 0. 05 ）. （2） The transwell results showed that the migrated VSMCs were significantly higher in thrombin group than in control group （ 1 337 ± 162 vs. 99 ±26, P 〈 0.01 ） , which could be significantly reduced by pretreatment with quercetin （ 926 ± 111, P 〈 0.05 ） , but still significantly higher than control group （ P 〈 0. 01 ）. （ 3 ） The protein expressions of phospho- extraeellular signal regulated kinase 1/2 and phosphor-p38 were significantly upregulated in thrombin groupcompared to control group （ both P 〈 O. 05 ）, which could be significantly downregulated by pretreatment with quercetin （P 〈 O. 05）. Conclusion Quercetin can effectively attenuate thrombin-induced vascular smooth muscle cells proliferation and migration, possibly through inhibiting the phosphorylation of extracellular signal regulated kinase 1/2 and p38 pathway.