莱菔硫烷对H9C2心肌细胞缺氧/复氧损伤的保护作用

摘要

Objective:To investigate the protective effects of sulforaphane (SFN )on hypoxia-reoxygenation (H/R)injury of myocardial cells. Methods:H9C2 cells were randomly divided into 3 groups:Normal control group,H/R group (cells subjected to hypoxia-reoxygenation)and SFN pretreatment group (after pretreatment by 10 μmol/L SFN for 12 h, hypoxia-reoxygenation was performed).The cell morphology and the degree of apoptosis were observed by inverted microscope. The survival rate of H9C2 cells in each group was detected by CCK8 method,and the protein expressions of HO-1 ,NQO1 ,Bcl-2 and Bax was detected by western blot. Results:Compared with the normal control group,the survival rates of cells in H/R group and SFN pretreatment group were significantlydecreased (all P<0.05),and the protein expressions of HO-1,NQO1, Bcl-2 and Bax, and the Bcl-2/Bax were increased (all P<0.05).The growth of cells in SFN pretreatment group were better than that in H/R group.The survival rate of cells in SFN pretreatment group was significantly increased compared with the H/R group [(76.00±0.52)% vs.(55.73±0.43)%,P<0.01].The protein expressions of HO-1(3.24±0.01 vs.1.86±0.01,P<0.01),NQO1(1.67±0.01 vs.0.95±0.01,P<0.01)and Bcl-2 (0.70±0.00 vs.0.48±0.01,P<0.01),and the Bcl-2/Bax(1.22±0.01 vs.0.56± 0.00,P<0.01)were increased,while the protein expressions of Bax were significantly decreased(0.57 ±0.00 vs.0.85± 0.01;P<0.01 )in SFN pretreatment group compared with the H/R group. Conclusion:SFN has protective effects on hypoxia-reoxygenation injury of H9C2 myocardial cells,which may be related to the promotion of HO-1 ,NQO-1 and Bcl-2 protein expression and the inhibition of Bax protein expression.%目的:探讨莱菔硫烷(sulforaphane,SFN)对H9C2心肌细胞缺氧/复氧损伤的保护作用.方法:H 9 C2心肌细胞随机分为3组:正常对照组、缺氧/复氧组(H/R组)和SFN预处理组(10μmol/L SFN预处理细胞12 h后,进行缺氧/复氧处理).采用倒置显微镜观察各组细胞形态及凋亡程度,CCK8法检测各组H9 C2心肌细胞存活率,Western blot法检测血红素氧化酶1(HO-1)、醌氧化还原酶1(NQO1)、Bcl-2和Bax蛋白的表达水平.结果:与正常对照组相比,H/R组和SFN预处理组心肌细胞存活率均降低(P均<0.01),HO-1、NQO1、Bcl-2和Bax蛋白表达水平及Bcl-2/Bax均升高(P均<0.01).与H/R组相比,SFN预处理组的细胞生长状态较好,细胞存活率明显提高[(76.00±0.52)%对(55.73±0.43)%,P<0.01],HO-1(3.24±0.01对1.86±0.01,P<0.01)、NQO1(1.67±0.01对0.95±0.01,P<0.01)和Bcl-2(0.70±0.00对0.48±0.01,P<0.01)蛋白表达水平及Bcl-2/Bax(1.22±0.01对0.56±0.00,P<0.01)均明显升高,Bax蛋白表达水平明显降低(0.57±0.00对0.85±0.01,P<0.01).结论:SFN对H9C2心肌细胞缺氧/复氧损伤有保护作用,可能与促进HO-1、NQO1、Bcl-2蛋白表达,抑制Bax蛋白表达有关.