衢枳壳不同组分体外降糖活性研究及4种黄酮组分含量分析

Study on the Hypoglycemic Activity of Different Components of Quzhiqiao in Vitro and Analysis of Four Flavonoids

关键词:  衢枳壳  黄酮类化合物  α-葡萄糖苷酶  HepG2  葡萄糖消耗

Key words:  Quzhiqiao  flavonoids  α-glucosidase  HepG2  glucose consumption

摘要:

目的 研究衢枳壳不同极性组分体外降糖活性，并明确各组分中4种黄酮类化合物的含量。方法 采用系统溶剂法提取与分离衢枳壳中不同极性的黄酮组分，通过体外试验明确各组分对α-葡萄糖苷酶活性和HepG2细胞葡萄糖消耗的影响。采用Agilent Zorbax SB C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈-0.02%磷酸水溶液（20︰80）为流动相，流速1.0 mL·min^-1，检测波长280 nm，柱温35℃，外标法测定衢枳壳各组分中芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷的含量。结果 衢枳壳各组分均具有α-葡萄糖苷酶抑制作用，抑制强弱顺序为正丁醇组分[IC₅₀：（0.033±0.010） mg·mL^-1] > 甲醇组分[IC₅₀：（0.092±0.006） mg·mL^-1] > 乙酸乙酯组分[IC₅₀：（0.170±0.014） mg·mL^-1] > 氯仿组分[IC₅₀：（0.509±0.070） mg·mL^-1]；衢枳壳甲醇组分和正丁醇组分能够显著促进HepG2细胞葡萄糖的消耗，差异具有统计学意义（P<0.05）。含量测定方法经方法学认证，各项参数均符合要求。经分析发现：衢枳壳各组分中均含有芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷等黄酮类化合物，且正丁醇组分中各黄酮成分含量最高。结论 衢枳壳各组分均具有α-葡萄糖苷酶抑制作用，且随着各组分中黄酮类化合物含量增高作用增强。同时，含有较高黄酮类组分含量的衢枳壳组分具有促进HepG2细胞葡萄糖消耗的作用，如甲醇、正丁醇和乙酸乙酯组分。

Abstract:

OBJIECTIVE To study the hypoglycemic activity of different components of Quzhiqiao in vitro and clarify the content of 4 flavonoid compounds in each component. METHODS The flavonoids with different polarities were extracted and separated by system solvent method, their α-glucosidase inhibitory activity were demonstrated by inhibitory test of α-glucosidase in vitro, and their effects on the glucose consumption of HepG2 cells were also investigated. An HPLC method was established with an Agilent Zorbax SB C₁₈ column (250 mm×4.6 mm, 5 μm), acetonitrile-0.02% phosphoric acid solution (20:80) as the mobile phase, the flow rate was 1.0 mL·min^-1, the detection wavelength was 280 nm, and the column temperature was 35℃. Finally, the content of rutinine, naringin, hesperidin and neohesperidin in different components of Quzhiqiao were determined by external standard method. RESULTS The components of Quzhiqao had α-glucosidase inhibitory effects, their order of inhibitory activity was:n-butanol component[IC₅₀:(0.033±0.010) mg·mL^-1] > methanol component[IC₅₀:(0.092±0.006)mg·mL^-1] > ethyl acetate component[IC₅₀:(0.170±0.014)mg·mL^-1] > chloroform components[IC₅₀:(0.509±0.070)mg·mL^-1]. The methanol component and n-butanol component of Quzhiqiao could significantly promote the consumption of glucose in HepG2 cells and the differences were statistically significant (P<0.05). Through methodological certification, the parameters of content determination method were all in line with the requirements. And the experiment found that each component of Quzhiqiao was all composed of rutininenaringin, naringin, hesperidin and new hesperidin, and the n-butanol component had the highest content of flavonoids. CONCLUSION Each component of Quzhiqiao all have good α-glucosidase inhibitory activity, and the inhibitory effect is enhanced with the increase of the concentration of flavonoids in the components. Simultaneously, the higher content of flavonoids such as methanol component, n-butanol component and the ethyl acetate component all have the effect of promoting the glucose consumption of HepG2 cells.