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目的探讨冠心平片含药血清抗H2O2诱导血管内皮细胞(vascular endothelialcells,VEC)凋亡作用及对核因子-κB(nuclear factor kappa B,NF-κB)蛋白表达的影响。方法将大白兔随机分为正常组(生理盐水,10mL/kg)、维拉帕米组(0.02g/kg,10mL/kg)、冠心平小剂量组(2.8g/kg,10mL/kg)、冠心平中剂量组(5.6g/kg,10mL/kg)、冠心平大剂量组(11.2g/kg,10mL/kg),每组3只。灌胃给药,连续3天,最后1天灌胃2次,间隔2h,末次给药后1h,耳缘静脉采血,放置1h,2500r/min离心30min,吸取血清,56℃、30min灭活,制备含药血清。采用100μmol/LH2O2作用于对数生长期的VEC建立细胞凋亡损伤模型。造模后分为6组,每组5个样本:正常组(10%空白血清培养液)、模型组(10%空白血清培养液+100μmol/LH2O2)、维拉帕米组(10%维拉帕米血清培养液+100μmol/LH2O2)、冠心平低剂量组(10%低剂量冠心平血清培养液+100μmol/LH2O2)、冠心平中剂量组(10%中剂量冠心平血清培养液+100μmol/LH2O2)、冠心平高剂量组(10%高剂量冠心平血清培养液+100μmol/LH2O2)。采用流式细胞术检测VEC凋亡率,采用Westernblot检测NF-κB蛋白表达。结果模型组VEC凋亡率(9.00%±1.18%)、NF-κB蛋白表达(0.39%±0.06%)较正常组升高(P<0.01);与模型组比较,维拉帕米组(6.00%±0.18%)及冠心平高剂量组(5.30%±0.08%)、中剂量组(6.83%±0.51%)VEC凋亡率明显降低,各给药组NF-κB蛋白表达显著降低(维拉帕米组:0.28%±0.03%,冠心平低剂量组:0.33%±0.03%,冠心平中剂量组:0.30%±0.03%,冠心平高剂量组:0.28%±0.04%,P<0.01,P<0.05)。结论冠心平片可以拮抗H2O2诱导VEC的凋亡效应,其机制可能与调节NF-κB有关。
Objective To investigate the effects of Guanxin Tablet-containing serum on H2O2-induced apoptosis of vascular endothelial cells (VECs) and the expression of nuclear factor-kappa B (NF-κB). Methods The rabbits were randomly divided into normal group (10mL / kg), verapamil group (0.02g / kg, 10mL / kg) Coronary heart flat dose group (5.6g / kg, 10mL / kg), Guanxintong high dose group (11.2g / kg, 10mL / kg), three in each group. Gavage for 3 consecutive days, the last 1 day gavage 2 times, interval 2h, 1h after the last administration, blood collection of the ear marginal veins, place 1h, 2500r / min centrifugation 30min, serum, 56 ℃, Preparation of drug-containing serum. The apoptotic model of VEC was established by using 100μmol / LH2O2 in logarithmic growth phase. The rats in the model group (10% blank serum medium + 100μmol / LH 2 O 2), verapamil group (10% Vera (10% lower dose of Guanxinping serum + 100μmol / LH2O2), Guanxinping medium dose (10% of medium dose of Guanxinping serum + 100μmol / L H2O2), low dose of Guanxinping (10% LH2O2), Guanxianping high dose group (10% high dose Guanxinping serum + 100μmol / LH2O2). The apoptosis rate of VEC was detected by flow cytometry. The protein expression of NF-κB was detected by Western blot. Results The apoptosis rate of VEC in model group (9.00% ± 1.18%) and the expression of NF-κB protein (0.39% ± 0.06%) were higher than those in normal group (P <0.01) % ± 0.18%) and Guanxiao Ping high dose group (5.30% ± 0.08%) and medium dose group (6.83% ± 0.51%). The apoptosis rate of VEC in each group was significantly decreased Pam group: 0.28% ± 0.03%, Guanxintong low dose group: 0.33% ± 0.03%, Guanxian Ping dose group: 0.30% ± 0.03% P <0.05). Conclusion Guanxin Tablet can antagonize the H2O2-induced apoptosis of VEC, and its mechanism may be related to the regulation of NF-κB.