TRPC通道阻滞剂SKF96365对缺氧复氧诱导新生大鼠肥大心肌细胞凋亡的影响

目的探讨TRPC通道阻滞剂SKF96365对缺氧复氧诱导肥大心肌细胞凋亡的影响。方法①HP组，血管紧张素Ⅱ刺激体外培养的新生鼠心室肌细胞构建肥大心肌细胞模型，用5％CO2＋95％N2培养条件下进行缺氧6h，然后恢复5％C02＋21％02的条件复氧3h。②HP—SKF组，在缺氧前加入SKF96365，其他同肥大组。③CON组，在整个实验过程加入生理盐水代替AngⅡ。用AnnexinVFITC和PI双染色检测细胞凋亡；Westernblot法检测TPRCl和胞衬蛋白的表达。结果HP组心肌细胞的凋亡率明显高于CON组和HP—SKF组。CON组TRPCI的表达低于HP组和HP—SKF组。HP组剪切的胞衬蛋白率与完整的胞衬蛋白比值高于CON组和HP—SKF组。结论SKF96365能减低缺氧复氧诱导的肥大心肌细胞的凋亡。

EFFECT OF TRPC CHANNELS BLOCKER ON HYPOXIA/REOXYGENATION-INDUCED APOPTOSIS OF HYPERTROPHIC CARDIOMYOCYTES

Objective To investigate the effect of SKF96363, a blocker of canonical transient receptor potential （TRPC） channels, on hypoxia/reoxygenation- induced apoptosis of hypertrophic cardiomyocytes. Methods Three groups were divided. （1） HP group： The hypertrophy of cardiomyocytes was induced by Ang Ⅱ for 48 hrs after ventricular myocytes of neonatal rats were isolated and cultured. Then cardiomyocytes were cultured under the hypoxic condition （5%CO2 ＋ 95%N2） for 6 hrs and under reoxygenic condition （5%CO2 ＋21%O2） for 4 hrs. （2）HP- SKF group： The treatments were the same as those of HP group except for adding SKF96365 before hypoxia. （3） CON group： The processes were like those of HP group except that normal saline was added instead of Ang Ⅱ. The apoptosis rates were assessed by the method of annexin V FITC/PI. The expressions of TRPC1 and α- fodrin were measured .by the method of Western blot. Results Apoptosis rates in HP group were higher than those in HP- SKF group and in CON group. The expressions of TRPC1 in CON group were lower than those in HP group and in HP- SKF group. The ratios of cleaved α-fodrin to noncleaved α-fodrin in HP group were higher than those in the CON group and in HP- SKF group. Conclusion SKF96365 can reduce the hypoxia/reoxygenation - induced apoptosis of hypertrophic cardiomyocytes.