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  

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

作　　者：石玥[1] 梁晓春[1] 张宏[2] 王普艳[1] 赵丽[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院中医科转化医学中心,北京100730 [2]中国医学科学院北京协和医学院基础医学研究所细胞中心,北京100005

出　　处：《基础医学与临床》2012年第9期1059-1063,共5页Basic and Clinical Medicine

基　　金：北京市自然科学基金(7082077)

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48 h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)检测细胞PARP-1(89 ku)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(89 ku)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。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. Methods SD rats were divided into normal control group （dis- tilled water） ,JMT group and vitamin C 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 （50 mmol/L glucose medium, JMT group （ JMT-medica-ted serum） and vitamin C （VC） group （ VC- medicated serum）. Schwann cells cultured in DMEM were used as the normal control. After 48 h culturing, the levelof 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. Results 1 ） Compared with high glu- cose 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. 2）Compared with high glucose group,the expression of PARP-1 （89 ku） in Schwann cells cultrued in JMT group decreased signifi- cantly（P 〈 0.01 ）. The expression of JMT group was also much lower than that of V C group （P 〈 0.01 ）. Conclu- sions The medicated serum of JMT down-regulates the expression of ROS and PARP-1 of Schwann cells cultured in high glucose medium and reduces the oxidative DNA damage.

关 键 词：雪旺细胞 高糖 氧化应激损伤 PARP-1 筋脉通 

分 类 号：R285.5[医药卫生—中药学]