银耳多糖抗心肌细胞凋亡作用的实验研究  

Anti-apoptosis effect of tremella polysaccharides on cardiomyocytes

作　　者：曲丹[1] 蔡东联[1] 张浩[2] 乔晓萍[1] 何芳[1] 王莹[1] 林宁[1] 

机构地区：[1]第二军医大学长海医院营养科,上海200433 [2]第二军医大学长海医院胸心外科,上海200433

出　　处：《第二军医大学学报》2009年第4期383-386,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30672733)~~

摘　　要：目的:从细胞及动物整体水平研究银耳多糖(tremella polysaccharides,TP)抗心肌细胞凋亡的作用。方法:采用醇提法提取精制TP,气相色谱法检测TP纯度。对乳鼠心肌细胞原代培养后随机分为正常对照组(A组),凋亡对照组(B组)和TP预处理组(T组),72 h后进行形态学观察和锥虫蓝摄取率测定。采用流式细胞术检测各组心肌细胞凋亡指数。在动物体内实验中,选择清洁级ICR纯系小鼠100只,按体质量随机分为5组:阴性对照组(N组)每天腹腔注射生理盐水,其余4组每天腹腔注射120 mg/kg D-半乳糖以建立小鼠衰老模型,同时阳性对照组(C组)每日以生理盐水灌胃,L、M、H组分别以每日100、200、400 mg/kg TP灌胃。8周后处死小鼠,获取心肌标本进行组织学研究和生化检测。结果:培养的各组乳鼠心肌细胞锥虫蓝摄取率:B>C>A(P<0.01);心肌细胞凋亡指数:B>C>A(P<0.01)。动物实验中,与N组相比,阳性对照组(C组)及各TP干预组心肌均无明显病理学改变。TUNEL法检测心肌细胞凋亡指数显示:H<M<C(P<0.05)。心肌组织匀浆检测结果显示:H、M组MDA和LP含量低于C组(P<0.05),且H组低于M组(P<0.05);各TP干预组GSH-Px和SOD活性均高于C组(P<0.05),而H组活性最高(P<0.05)。结论:银耳多糖能抑制氧化损伤诱导体外培养乳鼠心肌细胞凋亡的发生,对心肌细胞具有保护作用。对D-半乳糖致衰老模型小鼠心肌细胞具有抗凋亡和抗氧化作用,且这种作用具有剂量相关性。Objective：To observe the anti-apoptosis effect of tremella polysaccharides （TP） on cardiomyocytes by in vitro and in vivo experiments. Methods： TP was extracted by dehydrated alcohol and the TP purity was measured by gas chromatography. The cultured cardiomyocytes of the neonatal rats were randomly assigned to normal group （Group A）, apoptosis-induced group （Group B） and TP-pretreatment group （Group T）. The morphology and uptake rate of trypan blue were studied after 72 h. The apoptosis index of cardiomyocyte was measured by flow eytometry. In the in vivo study, 100 ICR mice were randomly assigned to 5 groups after paired by body weight as following： Group N （negative control group, peritoneally injected by saline daily）; Group C （positive control group, peritoneally injected by D-galaetose and saline daily）; Group L, M, and H （peritoneally injected by D-galactose and intragastric administered with TP at 100, 200, 400 mg/day, respectively）. Mice were sacrificed for histology and biochemistry studies after 8 weeks. Results： Data of cultured cardiomyocytes were as follows： Uptake rate of trypan blue： B〉C〉A （P〈0.01）;apoptosis index：B〉C〉A （P〈0.01）. In the animal studies there were no obvious difference in myocardium pathology between groups. The order of the apoptosis indices of myocardium was H〈M〈C （P〈0.05）. Myocardium homogenate detection showed that MDA and LP in Group H, M were lower than those in group C （P〈0.05） ; MDA and LP in group H were lower than those in group M （P〈0.05） ;the activities of GSH-Px and SOD in all TP-pretreatment groups were higher than those in group C （P〈0.05）, and that in group H was the highest （P〈 0. 05）. Conclusion： TP can suppress the apoptosis of cardiomyocytes induced by oxidative damage in vitro, indicating a protective effect on cardiomyocytes. Our results suggest that TP has anti-apoptosis and anti-oxidation effect on cardiomyocytes in D-galaetose induced aging mice and the effect is

关 键 词：银耳多糖 细胞凋亡 心肌细胞 抗氧化 

分 类 号：R286.7[医药卫生—中药学]