穿山龙米曲霉固体发酵物总皂苷的抗肿瘤活性研究  

Study on Antitumor Activity of Total Saponins from Dioscorea Aspergillus Oryzae Solid Fermentation

作　　者：陈航宇[1] 刘鹤[2] 娄婷婷[1] 何蕊[1] 贺丹彤 刘睿[2] 位鸿[1] CHEN Hangyu;LIU He;LOU Tingting;HE Rui;HE Dantong;LIU Rui;WEI Hong(The Affiliated Hospital to Changchun University of Chinese Medicine,Changchun 130021,China;Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学附属医院,长春130021 [2]长春中医药大学,长春130117

出　　处：《吉林中医药》2020年第4期526-529,545,共5页Jilin Journal of Chinese Medicine

基　　金：吉林省中医药科技项目(2017191);吉林省教育厅科技发展项目(JJKH20181275KJ)。

摘　　要：目的探讨穿山龙药材固体发酵对抗肿瘤活性的影响,并研究其抗肿瘤机制。方法 70%乙醇回流法对穿山龙药材以及穿山龙药材米曲霉固体发酵产物进行提取;D101树脂纯化;MTT法,双染法以及Western blotting探讨穿山龙总皂苷和穿山龙米曲霉发酵物总皂苷对HepG2的作用及机制。结果 MTT法结果显示穿山龙总皂苷和穿山龙米曲霉发酵物总皂苷均能使HepG2细胞存活率下降,呈现剂量依赖性;双染法结果表明不同浓度穿山龙总皂苷和穿山龙米曲霉发酵物总皂苷均可诱导HepG2细胞凋亡(P<0.001),凋亡比例明显呈浓度依赖性的升高;Western blotting结果显示,不同浓度穿山龙总皂苷和穿山龙米曲霉发酵物总皂苷处理的HepG2细胞,与空白对照组比较,Bcl-2、Bax、Caspase3蛋白表达水平无显著差异。结论穿山龙药材经过米曲霉固体发酵前后,均具有抗肿瘤活性,发酵没有使穿山龙药材抗肿瘤活性发生改变,均对HepG2细胞具有促进凋亡的作用,其机制可能为上调caspase-3的表达,对其上游Bax及Bcl-2的表达没有明显影响。Objective To investigate the effect of solid fermentation on tumor activity and its antitumor mechanism.Methods The solid fermentation products of aspergillus miltiorrhiza and aspergillus miltiorrhiza were extracted by 70% ethanol reflux method,purified by D101 resin,MTT,double staining and Western blotting to explore the effect and mechanism of total saponins of aspergillus paniculata and aspergillus paniculata fermentation on HepG2.Results The results of MTT assay showed that the survival rate of HepG2 cells decreased in a dose-dependent manner.The results of double staining showed that different concentrations of dioscorea or fermentation extract could induce apoptosis of HepG2 cells (P<0.001),and the proportion of apoptosis was significantly increased in a concentration-dependent manner.The results of Western blotting showed that there was no significant difference in the expression of Bcl-2,Bax,Caspase3 protein between HepG2 cells treated with different concentrations of dioscorea or fermentation extract and the blank control group.Conclusion Before and after solid fermentation of dioscorea aspergillus oryzae,the medicinal materials have antitumor activity always;Fermentation did not change the antitumor activity of dioscorea,and all of them could promote apoptosis of HepG2 cells.The mechanism may be to up-regulate the expression of caspase-3,but has no significant effect on the expression of bax and bcl-2 upstream.

关 键 词：穿山龙 固体发酵 促进HepG2凋亡 

分 类 号：R914[医药卫生—药物化学]