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

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.

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

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(distilled 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 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.Results 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.2)Compared with high glucose group,the expression of PARP-1(89 ku) 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).Conclusions 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.