南天竹总生物碱对三氧化二砷减毒作用及其LC-MS分析

目的:研究南天竹果实中总生物碱对三氧化二砷减毒的作用,并分析南天竹总生物碱部位的化学成分,为南天竹对三氧化二砷减毒的总生物碱有效部位研究提供研究基础。方法:采用三氧化二砷致急性损伤模型,依据生化指标及组织形态检测结果,比较三氧化二砷单用与总生物碱联用对实验鼠心、肝和肾功能的影响,进而评价南天竹总生物碱化学部位对三氧化二砷的减毒作用。采用Peakview(Version1. 2,AB SCIEX)软件对南天竹总生物碱部位质谱数据进行处理,依据化合物的相对分子质量(分子式),再结合二级碎片、色谱峰保留时间及文献信息等对总生物碱部位检测到的化合物进行确认。结果:单独给药组肌酸激酶(CK),乳酸脱氢酶(LDH),血清尿素氮(BUN),丙二醛(MDA),谷氨酸-丙酮酸氨基转移酶(ALT)和门冬氨酸氨基转移酶(AST)明显高于联合给药组和空白组,而其Na~+-K~+-三磷酸腺苷(ATP)酶,超氧化物歧化酶(SOD),过氧化氢酶(CAT)和血肌酐(SCr)消除率明显低于联合给药组和空白组;相对南天竹组,总生物碱组的LDH值的降低更多,而Na~+-K~+-ATP酶活性提高明显。联合用药组心脏细胞水肿情况有所改善,肾脏及肝脏损伤减轻。从南天竹生物碱部位鉴别出25个生物碱,已知化合物18个,未知7个,包括3种结构类型,其中以阿朴吗啡类生物碱居多。结论:总生物碱联合给药在心脏、肾脏、肝脏损伤程度较三氧化二砷单独给药有较明显的改善,提取纯化的总生物碱部位对三氧化二砷毒性具有减毒作用。南天竹组对三氧化二砷减毒作用较总生碱组总体具优势,但无显著差异,而总生物碱组对三氧化二砷致毒的心脏保护作用较南天竹组更优。南天竹总生物碱中的阿朴吗啡类生物碱如南天宁碱、南天竹碱等具有含量相对高和专属性较强的特点,可作为该部位质量控制方法的指标性成分。

Effect of Total Alkaloids from Nandina domfestica in Attenuating Arsenic Trioxide and Its LC-MS

Objective: To study the effect of total alkaloids of Nandina domestica in attenuating arsenic trioxide, and analyze chemical constituents of alkaloids extract of Nandina, in order to provide the theoretical basis for studying the effect of N. domestica in attenuating arsenic trioxide with alkaloids extract of N. domestica as effective fraction. Method: The model of acute liver injury induced by arsenic trioxide was used to compare the effects on heart and liver functions of mice between arsenic trioxide alone and total alkaloids of N.domestica combined with arsenic trioxide. The detoxification of total alkaloids on arsenic trioxide was evaluated based on biochemical parameters and pathological report. Peakview (Version1.2,AB SCIEX) software was used to process the mass spectrometry data of total alkaloids of N. domestica. The structures of determined compounds were identified by molecular weight of compound (molecular formula),secondary fragments,chromatographic peak retention and literature information. Result: Among biochemical indicators, creatine kinase(CK),lactate dehydrogenase(LDH),blood urea nitrogen(BUN),malondialdehyde(MDA),alanine aminotransferase(ALT) and amino transferase of aspartate(AST) of the arsenic trioxide group were increased,while elimination rates of Na⁺-K⁺-adenosine triphosphate(ATP),superoxide dismutase(SOD),catalase(CAT) and serum creatinine(SCr) were decreased,compared with those of the combination group. CK and LDH of the alkaloids extract group were more obviously increased than those of the N. domestica extract group,but with no remarkable difference. In histomorphometric examination,edema of mouse heart cells was improved,and some kidney and liver damages in rats were alleviated. Totally 25 alkaloids of alkaloid extract were identified. Among them,18 were known,and 7 were unknown,including 3 structural types,in which apomorphine alkaloids were mostly. Conclusion: Heart,kidney and liver damage degrees of the combination group were significantly alleviated compared with the arsenic trioxide group. The total alkaloids fraction extracted and purified have a significant attenuation in arsenic trioxide toxicity. The detoxification of total alkaloids extract was equal to that of N. domestica extract. Furthermore,apomorphine alkaloids, such as nantenine and domesticine, can be used as index components to establish a quality control method for total alkaloids.