玉郎伞两种黄酮单体对心肌细胞缺氧/复氧损伤的保护作用

Protection and its mechanism of two flavone morphons from Yulangsan on hypoxia-reoxygenation induced injury in myocardial cells

目的研究从玉郎伞(Yulangsan,YLS)中首次分离的两种黄酮单体对体外培养大鼠乳鼠心肌细胞缺氧/复氧损伤的保护作用,并初步探讨其作用机制。方法建立体外培养大鼠乳鼠心肌细胞缺氧/复氧损伤模型,倒置显微镜下观察加入YLS两种单体的含药血清(10%、5%、2.5%)后,各组缺氧/复氧心肌细胞形态学和搏动频率的变化;以MTT法检测各组细胞的存活率;用ELISA法测定心肌细胞Na+,K+-ATP酶、Ca2+,Mg2+-ATP酶活性及细胞培养上清液中总超氧化物歧化酶(T-SOD)、乳酸脱氢酶(LDH)、一氧化氮合酶(NOS)活性和丙二醛(MDA)含量。结果与模型组相比,YLS两种单体含药血清的高、中剂量均能明显增加心肌细胞的存活率,增强Na+,K+-ATP酶、Ca2+,Mg2+-ATP酶活性,降低细胞培养上清液LDH、NOS活性、MDA含量,提高T-SOD活性并呈剂量依赖性(P<0.05)。结论两种YLS单体对体外培养大鼠乳鼠心肌细胞缺氧/复氧损伤具有保护作用,其机制可能与清除自由基、抑制心肌细胞Ca2+超载有关。

Aim To observe the protective effects of two flavone morphons on hypoxia/reoxygenation（H/R） injury in myocardial cells and explore the mechanisms. Methods The hypoxia/reoxygenation injury model of cultured neonatal rat cardiomyocytes was developed. The cell morphology, the spontaneous beating, the survival rate, the content of LDH, NOS, T-SOD, MDA and the activity of Na^＋ , K^＋-ATP, Ca^2＋, Mg＆2 ＋ -ATP enzyme were determined in the cultured neonatal rat cardiomyocytes injuried by H/R. Results Compared with the model, preconditioning by two flavone morphons enhanced the spontaneous beating and the survival rate. Meanwhile, it increased the activity of T-SOD, Na^＋, K^＋-ATP, Ca^2＋, Mg^2＋-ATP enzyme and decreased the release of LDH, NOS, MDA （P 〈 0.05 ） in a dose-dependent manner. Conclusion The two flavone morphons have good effects on hypoxia/reoxygenation injury in cultured neonatal rat cardiomyocytes, which is related with scavenging free radicals, inhibiting Ca^2＋-overloading.