四种莲子心生物碱对H_2O_2诱导血管内皮细胞损伤的保护作用  

Protective effects of four alkaloids of embryo loti on H_2O_2-induced oxidative damage of vascular endothelial cells

作　　者：张玉玲[1] 杨光明[1] 李萍[1] 潘扬[1] 

机构地区：[1]南京中医药大学药学院,江苏南京210046

出　　处：《中国生化药物杂志》2015年第3期1-5,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金(81373295);江苏省高校优势学科建设工程资助项目(ysxk-2014)

摘　　要：目的探讨中药莲子心中4种生物碱莲心季铵碱(Lot)、莲心碱(Lien)、异莲心碱(Iso)和甲基莲心碱(Nef)对H2O2诱导人脐静脉内皮细胞株ECV304氧化损伤的保护作用。方法采用MTT法测定不同浓度Lot、Lien、Iso和Nef对正常和H2O2损伤ECV304细胞活性的影响,通过比色法测定并比较各组细胞一氧化氮(NO)及一氧化氮合酶(NOS)的水平。结果 Lot、Lien、Iso和Nef对正常ECV304细胞形态和活性均无影响;Lot为100μmol/L,Lien、Iso和Nef为0.1μmol/L时,对H2O2致损内皮细胞活性有较好的缓解作用,此时细胞活性分别为H2O2损伤组的112.8%、129.3%、125.6和118.2%(P<0.01或0.05)。4种生物碱在上述剂量时对H2O2致损内皮细胞形态的改善作用与阳性药卡托普利基本相近,除Lot的作用较弱外,Lien、Iso和Nef均可使受损内皮细胞的细胞间隙缩小,细胞彼此相连增强,细胞边界变清晰。另外,且4种莲子心生物碱均可提高H2O2致伤内皮细胞的NO浓度(P<0.05),增加损伤内皮细胞的NOS活性(P<0.05)。结论莲心季铵碱、莲心碱、异莲心碱和甲基莲心碱均对H2O2诱导内皮细胞损伤的有一定的保护作用,其作用机制可能是通过增加NOS生成提高血管内皮细胞释放NO,从而发挥保护内皮的功能。Objective To observe protective effects of four active liposoluble alkaloids of a Chinese herb, lotusine （Lot）, liensinine （Lien）, isoliensinine （Iso） and neferine （Nef） of embryo loti （the green embryo）, against H2 O2-induced oxidative damage on human umbilical vascular endothelial cell ECV-304.Methods The protective effects of Lot, Lien, Iso and Nef on the survival of normal and oxidatively damaged ECV 304 cells were studied by cell morphology observation and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium （MTT） assay.The levels of nitric oxide （NO） and nitric oxide synthase （ NOS） were measured using colorimetric assay.Results Lot, Lien, Iso and Nef did not affect cell morphology and cell viability of normal ECV 304 cells.The survival of oxidative damaged vascular endothelial cells was rescued by incubating with Lot at 100μmol/L, and Lien, Iso and Nef at 0.1 μmol/L.The proliferative activity of medicated groups increased to 112.8%, 129.3%, 125.6 and 118.2%, respectively （P〈0.01 or 0.05）, relative to that of the group with H2O2 induced oxidative damage.The four alkaloids restrained oxidative injury of endothelial cells induced by H2 O2 and the protective influences were similar with captopril, which served as a positive control.Each alkaloid except Lot reduced intercellular space and increased the connections of oxidative damaged cells, concomitant with more recognizable cell borders.Lien, Iso and Nef also increased the concentration of NO （ P〈0.05 ） .Besides, all of the four alkaloids activated NOS in damaged vascular endothelial cells （ P〈0.05 ） . Conclusion The four alkaloids of embryo loti, especially Lien, Iso and Nef, have certain protective effects against H2 O2-induced oxidative damage on vascular endothelial cells.The protective mechanism may be promotion of NO release through the increase of NOS production.

关 键 词：莲心季铵碱 莲心碱 异莲心碱 甲基莲心碱 血管内皮细胞 H2 O2诱导损伤 

分 类 号：R932[医药卫生—生药学]