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目的:研究淫羊藿主要成分淫羊藿苷对ConA(刀豆蛋白)致体外BRL-3A细胞(大鼠肝细胞)损伤的影响和作用机制。方法:体外培养BRL-3A细胞,分别加入25、50、100μmol/L的淫羊藿苷24h,再以100μg/ml的ConA诱导损伤24h,MTT法检测BRL-3A细胞的存活率,考察细胞上清液中谷草转氨酶、谷丙转氨酶、超氧化物歧化酶、丙二醛、谷胱甘肽过氧化物酶含量,以荧光探针DCFH-DA检测细胞内活性氧含量,免疫细胞化学方法检测肝细胞Bax、Bcl-2表达水平。结果:淫羊藿苷显著提高ConA刺激后BRL-3A的存活率,25μmol/L、50μmol/L、100μmol/L淫羊藿苷均明显降低谷草转氨酶的含量;100μmol/L淫羊藿苷明显降低谷丙转氨酶的含量,并明显升高超氧化物歧化酶、谷胱甘肽过氧化物酶的活性,降低丙二醛、活性氧含量,增强Bcl-2的表达,降低Bax的表达。结论:淫羊藿苷能明显改善ConA诱导的BRL-3A细胞损伤,作用机制与淫羊藿苷抗氧化相关。
Objective: To study the effect and mechanism of Icariin, a major component of Epimedium, on the injury of BRL-3A cells (rat hepatocytes) induced by ConA (concanavalin) in vitro. Methods: BRL-3A cells were cultured in vitro. The cells were treated with 25, 50 and 100μmol / L icariin for 24h, and then treated with 100μg / ml ConA for 24 hours. The survival rate of BRL-3A cells was measured by MTT assay Serum aspartate aminotransferase, alanine aminotransferase, superoxide dismutase, malondialdehyde, glutathione peroxidase content, fluorescent probe DCFH-DA detection of intracellular reactive oxygen species, immunocytochemical detection of liver Cell Bax, Bcl-2 expression levels. Results: Icariin significantly increased the survival rate of BRL-3A after ConA stimulation. 25μmol / L, 50μmol / L, 100μmol / L icariin significantly decreased the content of aspartate aminotransferase; 100μmol / L icariin significantly decreased Alanine aminotransferase, and significantly increased superoxide dismutase, glutathione peroxidase activity, lower malondialdehyde, reactive oxygen species, increased Bcl-2 expression, decreased Bax expression. CONCLUSION: Icariin can significantly improve the injury induced by ConA in BRL-3A cells. The mechanism is related to the antioxidation of icariin.