生物资源

2018, v.40;No.161(03) 246-251

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板桥党参多糖提取工艺优化及对肝癌细胞HepG 2的抑制
Polysaccharide extraction process optimization of Banqiao Codonopsis and its inhibition effect on human hepatoma cell HepG2

张骏;要辉;张崇;潘洁;张瑜;梁华丽;
ZHANG Jun;YAO Hui;ZHANG Chong;PAN Jie;ZHANG Yu;LIANG Huali;Pharmacy Department,Xiangyang No.1 People's Hospital,Hubei University of Medicine;School of Pharmacy,Hubei University of Chinese Medicine;Cardiology Department,Xiangyang Central Hospital;

摘要(Abstract):

本文采用星点设计—效应面优化法和正交设计优化板桥党参多糖的提取工艺,并对两种工艺进行比较,同时初步考察了板党多糖对肝癌细胞Hep G2细胞的细胞毒作用。本实验的两种方法均以粗多糖得率为因变量,以液料比、提取时间、提取温度为自变量对提取工艺进行系统考察。星点设计优选的最佳工艺为提取时间154.6 min,料液比1∶12.3(m/v),提取温度91.5℃,板党多糖得率为(18.67±1.23)%;正交设计优选的最佳工艺为料液比1∶12(m/v),提取时间3.0 h,提取次数2次,板党多糖提取率为(15.49±1.36)%;板党多糖对Hep G2细胞细胞毒作用采用噻唑蓝(MTT)法进行检测,结果 24 h抑制Hep G2细胞的IC_(50)值为346.322μg/m L,48 h IC_(50)值为325.609μg/m L。同时检测了板党多糖对Hep G2细胞的凋亡诱导作用,结果显示浓度为400μg/m L时对细胞的凋亡诱导作用最强。实验结果表明两种实验设计方法所优化工艺条件基本相近,但是星点设计—效应面优化法精度更高,可用于指导制剂生产;细胞毒作用的结果表明板党多糖具有抑制Hep G2细胞增殖的能力。
In this paper,the central composite design-response surface method and orthogonal design were used to optimize the extraction process of Banqiao Codonopsis polysaccharides,and the two processes were compared. At the same time,the cytotoxicity of the polysaccharide on human hepatoma cell Hep G2 was investigated. Both methods of this experiment took the yield of crude polysaccharide as the dependent variable,the liquid-material ratio,the extraction time,and the extraction temperature as the independent variables to conduct a systematic inspection of the extraction process. The final optimal design for central composite design was the extraction time of 154.6 minutes,the ratio of material to liquid of 1∶12.3,and the extraction temperature of 91.5 °C;the yield of Bantang polysaccharide was(18.67±1.23)%.The optimum technology for orthogonal design was the ratio of material to liquid of 1∶12,the extraction time of 3.0 h,and the extraction times of 2;the polysaccharide extraction rate was(15.49±1.36)%. The cytotoxicity of extracted polysaccharide on Hep G2 cells was detected by methyl tetrazolium(MTT)assay. The results showed that the IC_(50) value of Hep G2 cells was inhibited at 346.322 μg/m L for 24 h,and the IC_(50) value at 48 h was 325.609 μg/m L. At the same time,the apoptosis induction of Hep G2 cells was tested. The results showed that extracted polysaccharide at 400 μg/m L had the strongest induction of apoptosis.These results indicate that the experimental conditions optimized by the two experimental methods are basically similar,but the central composite design-response surface method is more accurate and can be used to guide the production of the preparation. The results of cytotoxicity show that the polysaccharide can inhibit the proliferation of Hep G2 cells.

关键词(KeyWords): 板桥党参;星点设计—效应面法;多糖得率;抑制;噻唑蓝(MTT)
Banqiao Codonopsis;central composite design-response surface method;polysaccharide yield;inhibi tion;methyl tetrazolium(MTT)

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基金项目(Foundation): 国家自然科学基金青年基金(8170131364)

作者(Author): 张骏;要辉;张崇;潘洁;张瑜;梁华丽;
ZHANG Jun;YAO Hui;ZHANG Chong;PAN Jie;ZHANG Yu;LIANG Huali;Pharmacy Department,Xiangyang No.1 People's Hospital,Hubei University of Medicine;School of Pharmacy,Hubei University of Chinese Medicine;Cardiology Department,Xiangyang Central Hospital;

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