基础医学与临床 ›› 2012, Vol. 32 ›› Issue (9): 1059-1063.

• 研究论文 • 上一篇    下一篇

筋脉通含药血清降低高糖培养大鼠雪旺细胞活性氧水平及PARP-1蛋白表达

石玥1,梁晓春1,张宏2,王普艳1,赵丽1   

  1. 1. 中国医学科学院 北京协和医学院 北京协和医院
    2. 中国医学科学院基础医学研究所;北京协和医学院基础学院
  • 收稿日期:2011-10-31 修回日期:2011-12-25 出版日期:2012-09-05 发布日期:2012-08-28
  • 通讯作者: 梁晓春 E-mail:xcliang@vip.sina.com
  • 基金资助:
    筋脉通对糖尿病周围神经病变氧化应激致细胞凋亡的影响

Effects of Chinese herbal medicine Jinmaitong-medicated serum on the ROS level and expression of PARP-1 of rat Schwann cells cultured in high-glucose medium

  • Received:2011-10-31 Revised:2011-12-25 Online:2012-09-05 Published:2012-08-28
  • Contact: LIANG Xiao-chun E-mail:xcliang@vip.sina.com

摘要: 目的 研究筋脉通含药血清对体外高糖培养雪旺细胞活性氧(ROS)水平及多聚(ADP-核糖)聚合酶-1(PARP-1)蛋白表达的影响。方法 30只雄性SD大鼠随机分为3组,分别灌胃筋脉通(JMT)、维生素C(VC)或蒸馏水制备含药血清和对照血清。取新出生大鼠的双侧坐骨神经用于制备雪旺细胞,分为高糖组、JMT组(加入筋脉通含药血清)、VC组(加入维生素C含药血清)及正常对照组。培养48h后, 采用激光扫描共聚焦显微技术检测细胞内二氯荧光黄(DCF)的荧光强度而测得细胞内ROS水平;采用免疫印迹法(Western blot)的方法检测细胞PARP-1(89ku)的蛋白表达。结果 1)高糖组雪旺细胞ROS-DCF荧光值(86.59±8.14)显著高于正常值(P<0.01);筋脉通含药血清组(44.58±8.67)与维生素C含药血清组(46.12±8.80)均显著低于高糖组(P<0.01)。2)与正常组比较,高糖组雪旺细胞内PARP-1(89ku)含量(0.712±0.012)明显增加(P<0.01);与高糖组比较,筋脉通含药血清组含量(0.307±0.025)明显降低(P<0.01),且明显优于维生素C组(0.594±0.017)(P<0.01)结论 筋脉通含药血清能显著减少ROS生成,降低PARP-1的活性和酶解,减轻细胞DNA氧化损伤。

关键词: 雪旺细胞, 高糖, 氧化应激损伤, PARP-1, 筋脉通

Abstract: Objective To investigate the effects of medicated serum prepared by administration of Jinmaitong(JMT), a compound Chinese herbal medicine, on oxidative damage and poly ADP-ribose polymerase-1(PARP-1) of Schwann cells cultured in high-glucose medium, Method SD rats were divided into normal control group(distilled water),JMT(1.31g/(kg?d)) group and vitamin C(0.08g/(kg?d))group to prepare medicated serum. Bilateral sciatic nerves of new born SD rats were used to separate Schwann cells. Schwann cells cultured in high-glucose medium were divided into high glucose group (50mmol/L glucose medium,JMT group(JMT-medicated serum) and vitamin C(VC)group (VC-medicated serum).Schwann cells cultured in DMEM were used as the normal control. After 48h culturing, the level of ROS was measured by confocal laser scanning microscope with 2',7'-dichlorofluorescein(DCF) as a molecular probe and the expression of PARP-1 protein was detected by Western blot. Result: (1) Compared with high glucose group, the fluorescence intensities of ROS-DEC in Schwann cells cultured in JMT and VC groups were weaker significantly(P<0.01). There were no significant differences between these two treated groups(P>0.05). (2)Compared with high glucose group,the expression of PARP-1(89ku) in Schwann cells cultrued in JMT group decreased significantly(P<0.01).The expression of JMT group was also much lower than that of VC group(P<0.01).Conclusion: The medicated serum of JMT could down-regulate the expression of ROS and PARP-1 of Schwann cells cultured in high glucose medium, reduce the oxidative DNA damage.

Key words: Schwann cell, high glucose, oxidation damage, PARP-1, Jinmaitong capsule