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.
出处
《基础医学与临床》
CSCD
北大核心
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)
