摘要
目的:研究西洛他唑(Cilostazol,CIL)对乳鼠心肌细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的影响及机制。方法:①分离培养SD乳鼠心肌细胞,建立H/R模型。心肌细胞随机分4组:对照组;H/R组,缺氧2 h,复氧4 h;H/R+CIL组,缺氧前1 h予以CIL(10μmol.L-1)随即缺氧2 h,复氧4 h;H/R+CIL+Wort mannin(H/R+CIL+W)组,CIL处理前30 min予磷脂酰肌醇-3激酶(phosphatidylinositol-3 kinase PI3K)特异性抑制剂——渥曼青霉素(Wort mannin 0.1μmol.L-1);②检测各组培养液中乳酸脱氢酶、肌酸激酶同工酶含量,流式细胞术检测心肌细胞凋亡率,Western blot检测丝氨酸/苏氨酸蛋白激酶(Akt)及磷酸化丝氨酸/Akt(p-Akt)的表达。结果:CIL明显抑制H/R损伤导致的心肌细胞凋亡,并使p-Akt/Akt比值明显增加;Wort mannin可使p-Akt/Akt比值明显减少,抑制CIL的抗凋亡作用。结论:H/R损伤可导致心肌细胞凋亡,CIL可能通过PI3K-Akt途径发挥抗凋亡作用。
Objective：To investigate the protective effects of cilostazol on neonatal SD rat cardiomyocytes injury induced by hypoxia/reoxygenation and explore the mechanisms involved. Method： The cultured neonatal SD rat cardiomyoeytes were divided randomly into four groups：hypoxia/reoxygenation group（H/R） ：deoxygenation for 2 h and reoxygenation for 4 h ;hypoxia/reoxygenation＋ cilostazol group（ H/R＋ CIL） cilostazol （10 μmol·L^-1 ） was added lh before deoxygenation then deoxygenation for 2 h and reoxygenation for 4 h ;hypoxia/reoxygenation ＋ cilostazol＋Wortmannin group （ H/R＋ CIL＋ W） ： Wortmannin（0. 1 μmol ·L^-1 ）, a specific phosphatidylinositol-3 kinase（PI3K） inhibitor, added 30 min before cilostazol incubation. Activities of LDH and CK-MB in culture medium in each group were measured. Cardiocyte apoptosis were measured by flow cytometre. Expression of Serine/ threonine protein kinase （Akt） and phospho-Serine/threonine protein kinas（p-Akt） were detected hy Western blot. Result：Cilostazol significantly decreased the apoptotic index induced by hypoxia/reoxygenation and strongly increased p-akt/akt level. Wortmannin attenuated the antiapoptosis effects of cilostazol accompany with reduction of p-Akt/Akt level. Conclusion： Hypoxia/reoxygenation injury lead to cardiomyocyte apoptosis, Cilostazol inhibit apoptosis partly through PI3K/Akt pathway.
出处
《临床心血管病杂志》
CSCD
北大核心
2008年第10期775-778,共4页
Journal of Clinical Cardiology
