积雪草乙酸乙酯提取物对Aβ_(25-35)片段所致的PC12细胞损伤模型及SAMP8小鼠脑内SOD,GSH-Px的影响  

Effect of Ethyl Acetate Extract from Centella asiatica on PC12AD Cell Model and SOD,GSH-Px of SAMP8

作　　者：李霏 黄金兰[2] 崔雯[3] 张雯艳[3] 冯颖琴 郭尔楚 陈奔[3] 郭茜茜[3] 钟振国[3] 

机构地区：[1]广西桂东卫生学校,广西贺州543100 [2]徐州医学院,江苏徐州221004 [3]广西中医药大学,南宁530001

出　　处：《中国实验方剂学杂志》2014年第4期111-114,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81160512);广西自然科学基金项目(2010GXNSFD013050);广西创新研究团队项目(2011GXNSFF018006)

摘　　要：目的:研究积雪草乙酸乙酯提取物对β淀粉样蛋白25-35片段(Aβ25-35)所致的PC12细胞损伤模型及快速老化模型小鼠(SAMP8)脑内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的影响。方法:细胞以1×104个/mL密度接种,预防实验在接种24 h后同时给予含不同质量浓度的药物培养液和终浓度为10μmol·L-1的Αβ25-35溶液,治疗实验在接种24 h后给予终浓度为10μmol·L-1的Αβ25-35溶液,24 h后给予含不同质量浓度的药物培养液,作用72 h后采用MTT法检测积雪草乙酸乙酯提取物对Aβ25-35片段所造成的PC12细胞老年性痴呆(AD)损伤模型的预防和治疗作用;Hoechst 33258荧光显微镜染色法观察积雪草乙酸乙酯提取物作用Aβ25-35片段所致PC12细胞AD损伤模型的形态学变化,SAMP8)40只随机分为模型对照组、积雪草乙酸乙酯提取物高、中、低剂量组(以生药量计为40,20,10 g·kg-1)和石杉碱甲组(0.386 mg·kg-1),8只/组,连续ig给药2个月后,ELISA法测定积雪草乙酸乙酯提取物对SAMP8大脑组织SOD和GSH-Px的影响。结果:Aβ25-35片段所致的PC12细胞AD模型的预防和治疗实验中,不同浓度的积雪草乙酸乙酯提取物对Aβ25-35片段所致PC12细胞AD损伤有不同程度的保护作用;荧光显微镜观察到用药组的凋亡细胞数少于模型对照组与MTT的结果相符;ELISA结果显示除低剂量组外,其他各组的SAMP8大脑组织的SOD均高于模型对照组(P<0.05或P<0.01),中剂量组和石杉碱甲组SAMP8大脑组织的GSH-Px比模型对照组高(P<0.05或P<0.01)。结论:积雪草乙酸乙酯提取物在体外对由Aβ25-35片段所致的PC12细胞损伤有较好的保护和治疗作用,并能通过提高SAMP8脑内SOD,GSH-Px水平起到抗氧化,清除体内自由基而延缓衰老的作用。Objective： To explore the effects of Alzheirmer＇s disease （AD） model induced by β amyloid dismutase （SOD） and glutathione peroxidase （GSH-Px） ethyl acetate extract from Centella asiatica on PC12 cell 25-35 fragment （Aβ25-35 and the levels of superoxide in the brain of SAMP8. Method： The cell suspension（1 × 10^4 cell/mL） was seeded into 96-well plates for 24 h. For preventive trial, 10 ×mol .L-1 Aβ25-35 and drugs were added into 96-well plates. For treatment trail, drugs were added 24 h after 10 ×mol .L-1 Aβ25-35 was added into the cells. After 72 h, MTT assay was used to detect the prevention and treatment effect of AD model which made in PC12 cell induced by Aβ25-35 fragment after treated with ethyl acetate extract of C. asiatica, and fluorescence microscopy was applied to observe the morphological changes of the PC12 cells. For animal trail, forty senescence accelerated mouse-prone 8 （SAMP8） were divided into 5 groups. There were 8 mice in each group. Each mouse was intragastricly administrated for 2 months. The contents of SOD and GSH-Px in the brain of SAMP8 were determined by enzyme linked immunosorbent assay （ELISA）. Result： Different concentrations of ethyl acetate extract had different degrees of protection on PC12 cells, the results of fluorescence microscope was the same as MTT. ELISA demonstrated that the contents of SOD in high, mid-dosage ethyl acetate extract and Huperzine （Hup-A） groups were higher than that in control group, and the contents of GSH-Px treatment with mid-dosage ethyl acetate extract and Hup-A increased compared with control group. Conclusion： Ethyl acetate extracts of C. asiaticashows some degree of protective and therapeutic effects on the damage of PC12 cell caused by the Aβ25-35 fragments in vitro. Furthermore, ethyl acetate extracts of C. asiatica has antioxidant, scavenging oxygen free radicals effects which results in delaying senescence via enhancing the contents of SOD and GSH-Px.

关 键 词：积雪草乙酸乙酯提取物 PC12细胞株 Aβ25-35 快速老化模型小鼠SAMP8 超氧化物歧化酶 谷胱甘肽过 氧化物酶 

分 类 号：R285.5[医药卫生—中药学]