珠子参总皂苷通过促进Nrf2转位拮抗新生大鼠心肌细胞氧化应激损伤.docVIP

PAGE PAGE 7 珠子参总皂苷通过促进Nrf2转位拮抗新生大鼠心肌细胞氧化应激损伤 贺海波1，石孟琼2[通信作者] 石孟琼(1971-), 女, 实验师, 研究方向: 中药与心脑血管药理. 邮箱: shmq0212@126.com.，罗 涛3，陈良金2，周继刚3，张继红3，卢训丛2 （443002 [通信作者] 石孟琼(1971-), 女, 实验师, 研究方向: 中药与心脑血管药理. 邮箱: shmq0212@126.com. [摘要] 目的 观察珠子参总皂苷H2O2所致氧化应激损伤保护效应并探讨其机制。方法Wistar乳鼠心肌细胞原代培养，用H2O2建立氧化应激损伤模型，然后用珠子参总皂苷（100、200μg/ml）孵育24 h 后，观察细胞搏动频率，MTT 法检测珠子参总皂苷对细胞活力的影响，流式细胞仪和Hoechst33258 染色检测珠子参总皂苷对氧化应激诱导细胞凋亡及胞内ROS含量的影响，比色法测定培养液中乳酸脱氢酶（LDH）、肌酸激酶（CK）含量，同时检测培养液中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）的活性及丙二醛（MDA）含量，实时定量PCR技术检测珠子参总皂苷对SOD/GPX/CAT反应系统SOD1~ SOD3、CAT和GPX1基因表达的影响，Western blot检测珠子参总皂苷对细胞质和细胞核Nrf2蛋白表达的影响。结果 H2O2明显影响细胞活力和诱导凋亡(IC50为500μmol/L)，珠子参总皂苷（100、200μg/ml）处理能显著增加心肌细胞搏动频率和存活率（P＜0.01），抑制心肌细胞凋亡和改善细胞形态，降低细胞内ROS含量（P＜0.01）；减轻H2O2所致乳鼠心肌细胞培养液中 LDH、CK释放量，同时能够升高SOD、CAT和GSH-Px活性，降低MDA含量（P0.05，P0.01）；上调SOD/GPX/CAT反应系统基因表达水平和促进Nrf2核移位，增加心肌细胞核内Nrf2的表达水平（P0.05，P0.01）。结论 珠子参总皂苷抑制心肌细胞氧化应激损伤，与Nrf2抗氧化途径的激活和清除ROS有关。 [关键词] 竹节参总皂苷；过氧化氢；心肌细胞；转录因子Nrf2；氧化应激；保护机制 [中图法分类号] [文献标志码] A Protective effect of total saponin from Panacis majoris rhizoma on oxidative stress-induced injuries of neonatal rat cardiomyocytes Nrf2 translocation He Haibo1, Shi Mengqiong2, Luo Tao3, Chen Liangjin2, Zhou Jigang3, Zhang Jihong3, Lu Xuncong2 (1Hubei Key Laboratory of Natural Products Research and Development, 2 Experimental Center of Basic Medicine, Medical college, 3 Medical Department of Traditional Chinese Medicine Clinical Hospital, China Three Gorges University,Yichang,Hubei Province,443002,China） [Abstract] Objective To study the effect of total saponin from Panacis majoris rhizoma on oxidative stress-induced injuries of neonatal rat cardiomyocytes and its mechanism. Methods Oxidative stress-induced injury model was established with H2O2. Effects of total saponin from Panacis majoris rhizome(100, 200μg/ml) on beating rates of cardiomyocytes were monitored under inverted microscope, viability of cells and oxidative stress-induced apoptosis of cells was detected by MTT as