油菜蜂花粉活性成分对体外肝细胞损伤的保护作用

Hepatoprotective Effects of Bioactive Compounds from Rape Bee Pollen on L-02 Cells Injured by CCl4

摘要:

本文通过对油菜蜂花粉的分离纯化，得到槲皮素苷QMP、山奈酚苷KMG和KMP三个化合物。通过体外培养L-02细胞，建立四氯化碳损伤模型，采用MTT实验，来研究QMP、KMG和KMP抑制四氯化碳诱导L-02细胞损伤的作用。并选用丙二醛（MDA）、超氧化物歧化酶（SOD）和乳酸脱氢酶（LDH）作为特征性指标，从氧化应激角度探究保肝作用机制。结果表明，槲皮素苷QMP处理组（200 μg/mL）细胞相对存活率分别是模型组的2.64和2.66倍，说明QMP能够明显抑制四氯化碳对L-02细胞造成的损伤，山奈酚苷KMP和KMG抗L-02细胞的损伤作用不明显。QMP、山奈酚和槲皮素显著降低细胞内MDA含量，其中300 μm剂量组MDA含量分别为模型组的0.65、0.60、0.59和0.52倍；LDH漏出率分别降低了23.82%、27.69%、30.11%和32.77%；同时SOD含量分别提高了1.45、1.61、1.58和1.65倍。这些体外实验证明了QMP具有显著的体外抗氧化和保肝作用，且其抗氧化作用是保肝作用机制之一，为开发治疗肝损伤性疾病的药物提供了选择。

Abstract:

Quercetin-3-O-β-D-glucosyl-(2→l)-β-glucoside (QMP), kaempferol-3,4'-di-O-β-D-glucoside (KMG), and kaempferol-3-O- β-D-glucosyl-(2→l)-β-D-glucoside (KMP) were isolated and purified from rape bee pollen. A model of CCl4-induced liver injury was established via in vitro cultivation of L-02 cells, and the protective effects of QMP, KMG, and KMP on L-02 cells with CCl4-induced injury were studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The contents of malondialdehyde (MDA), superoxide dismutase (SOD), and lactic dehydrogenase (LDH) were used as the characteristic indicators to study the hepatoprotective effect in terms of oxidative stress. The relative cell survival rate of QMP-treated cells (200 μg/mL) was 2.64 and 2.66 times as much as that of the model group, respectively, suggesting that QMP can effectively prevent CCl4-induced liver injury in L-02 cells, but the hepatoprotective effects of KMG and KMP were not significant. QMP, kaempferol, and quercetin could significantly reduce intracellular MDA levels; the MDA levels of 300 μM dose groups were 0.65, 0.60, 0.59 and 0.52 times as much as that of the model group, respectively; the corresponding LDH leakage rates were reduced by 23.82%, 27.69%, 30.11%, and 32.77%, respectively; the SOD contents were increased by 1.45, 1.61, 1.58, and 1.65, respectively. The results suggest that QMP possesses significant in vitro antioxidant activity and a hepatoprotective effect, the antioxidant activity is one of the mechanisms underlying its hepatoprotective effect. This study provides a basis for developing drugs to treat diseases associated with liver injury.