摘要
目的研究羟基红花黄色素A(hydroxysafflor yellow A,HSYA)对心肌细胞模拟缺血/再灌注(simulated ischemia/reperfusion,SI/R)损伤的保护作用。方法采用H9c2心肌细胞建立SI/R损伤模型,检测心肌细胞存活率、乳酸脱氢酶(LDH)水平,TUNEL染色法检测细胞凋亡,Western blot检测Akt/e NOS信号通路蛋白的表达。结果与SI/R组比较,20μM HSYA预处理能显著提高SI/R损伤的H9c2细胞的存活率,(69.7±5.8)%vs.(79.9±5.8)%,差异有统计学意义(P<0.05)。与Control组相比,SI/R组心肌细胞LDH释放增加,(43.3±9.4)U/L vs.(129.5±11.9)U/L,差异有统计学意义(P<0.05)。与SI/R组相比,HSYA可显著降低LDH的释放,而加入PI3K的抑制剂LY29004后,这种作用消失。与Control组比较,SI/R损伤显著增加H9c2心肌细胞的凋亡率,差异有统计学意义(P<0.05);与SI/R组相比,SI/R+HSYA组细胞凋亡率显著下降,而加入LY29004后,逆转了HSYA的抑制凋亡的作用。与SI/R组相比,SI/R+HSYA组p-Akt蛋白的表达显著升高,差异有统计学意义(P<0.05),加入LY29004后p-Akt蛋白的表达被明显抑制。与SI/R组比较,SI/R+HSYA组p-e NOS蛋白的表达明显升高,差异有统计学意义(P<0.05),加入LY29004后p-e NOS蛋白的表达被明显抑制。结论羟基红花黄色素A能增强H9c2细胞活力,减少心肌细胞损伤,抑制心肌细胞凋亡,并通过激活Akt/e NOS信号通路发挥保护作用。
Objective To study the preventive effect of hydroxysafflor yellow A(HSYA) on simulated cardiomyocyte ischemia-reperfusion(SI/R) injury. Methods The model of SI/R injury was established by using H9c2 cardiomyocyte. The survival rate of cardiomyocyte and level of lactate dehydrogenase(LDH) were detected, apoptosis was detected by using TUNEL staining assay, and Akt/e NOS protein expressions were detected by using Western blot test. Resutls The survival rate of H9c2 cardiomyocyte with SI/R injury increased significantly in SI/R+HSYA group compared with SI/R group [(69.7±5.8) % vs.(79.9±5.8)%, P<0.05]. The release of LDH increased in SI/R group compared with control group [(43.3±9.4) U/L vs.(129.5±11.9) U/L, P<0.05]. HSYA decreased significantly the release of LDH, which disappeared after LY29004(PI3K inhibitor) added. The apoptosis rate of H9c2 cardiomyocyte increased significantly in SI/R group compared with control group(P<0.05). The apoptosis rate of H9c2 cardiomyocyte decreased significantly in SI/R+HSYA group compared with SI/R group, which was reversed after LY29004 added. The expression of p-Akt increased significantly in SI/R+HSYA group compared with SI/R group(P<0.05), which was significantly inhibited after LY29004 added. The expression of p-e NOS increased significantly in SI/R+HSYA group compared with SI/R group(P<0.05), which was significantly inhibited after LY29004 added. Conclusion HSYA can improve the viability of H9c2 cardiomyocyte, relieve cardiomyocyte injury, inhibit cardiomyocyte apoptosis, and play a protective role through activating Akt/e NOS signal path.
出处
《中国循证心血管医学杂志》
2015年第5期663-665,671,共4页
Chinese Journal of Evidence-Based Cardiovascular Medicine
