东北道地药材党参愈伤组织培养条件的优化及多糖的抗肿瘤活性研究  

Study on The Optimization of Callus Culture Conditions of Codadonopsis pilosula and the Antitumor Activity of its Polysaccharides

作　　者：高铭泽 田海燕 王祺瑶 徐贵国 凌娜 汲晨锋 GAO Mingze;TIAN Haiyan;WANG Qiyao;XU Guiguo;LING Na;JI Chenfeng(Pharmaceutical Engineering Technology Research Center,Harbin University of Commerce,Harbin 150076,China;School of Pharmocology,Harbin University of Commerce,Harbin 150076,China)

机构地区：[1]哈尔滨商业大学药物工程技术研究中心,黑龙江哈尔滨150076 [2]哈尔滨商业大学药学院,黑龙江哈尔滨150076

出　　处：《特产研究》2025年第1期129-135,共7页Special Wild Economic Animal and Plant Research

基　　金：黑龙江省中医药科研项目(ZYW2022-093)。

摘　　要：本研究旨在筛选出适于党参愈伤组织培养的影响因素,为党参组织培养体系的建立提供理论依据。通过对党参的不同部位诱导产生愈伤组织,采用正交试验法筛选获得诱导率最高的外植体、最佳外植体龄、最适培养基、最佳激素配比、最适温度、最适光照周期等影响因素,并对不同培养条件下党参愈伤组织多糖含量进行测定以及MTT法对党参多糖抗肿瘤活性进行研究。结果表明,叶片和茎段为外植体产生的愈伤组织最多,诱导率分别为90.67%和88%;多糖含量分别为127.54 mg/g和108.39 mg/g;28 d时叶片和茎段的诱导率达到最大值,分别为88.67%和84.67%,多糖含量为125.04 mg/g和104.22 mg/g;MS培养基明显优于其他几种培养基,诱导率达到92%,多糖含量为127.54 mg/g;在15~25℃时,诱导率急剧增加,并在25℃时诱导率达到最大值90.67%,多糖含量为162.45 mg/g;光照周期为6 h/d,愈伤组织诱导率为85.33%,多糖含量为102.45 mg/g。当激素组合为2,4-D 1 mg/L+6-BA 0.5 mg/L+NAA 1 mg/L,诱导率为98.67%,多糖含量最高为172.95 mg/g。此外,愈伤组织多糖对人乳腺癌MCF-7细胞和人宫颈癌HeLa细胞具有显著的生长抑制活性,作用48 h的IC50值分别为84.39 mg/L和164.78 mg/L。党参愈伤组织诱导的最佳外植体为叶片和茎段,最佳外植体龄为28 d,最佳培养基为MS+2,4-D 1 mg/L+6-BA 0.5 mg/L+NAA 1mg/L;25℃和6 h/d的光周期为其最适培养温度与光照条件。党参愈伤组织中提取的多糖具有显著的抗肿瘤活性。To screen out the suitable factors for callus culture of Codonopsis pilosula,and to provide theoretical basis for the establishment of C.pilosula tissue culture system.The optimal explants,explant age,culture temperature,light and culture medium of C.pilosula were selected from various influencing conditions of C.pilosula callus by orthogonal test,and the polysaccharide content under different culture conditions and the antitumor activity of C.pilosula polysaccharide were also studied by MTT assay.The optimal explants were leaves and stem segments,with induction rates of 90.67%and 88%,and the polysaccharide content of 127.54 mg/g and 108.39 mg/g,respectively.At 28 days,the induction rate of leaves and stem segments reached the maximum of 88.67%and 84.67%,and the polysaccharide contents were 125.04 mg/g and 104.22 mg/g,respectively.Moreover,MS medium is better than other media,with the induction rate of 92%and the polysaccharide content of 127.54 mg/g.Besides,the induction rate increased sharply from 15℃to 25℃,and reached a maximum of 90.67%at 25℃.The optimum light period was 6 h/d with the callus induction rate of 85.33%and the polysaccharide content of 102.45 mg/g.Moreover,under the hormone combination of 2,4-D 1 mg/L+6-BA 0.5 mg/L+NAA 1 mg/L,the induction rate was 98.67%and the highest pol-ysaccharide content was 172.95 mg/g.In addition,the callus polysaccharide of C.pilosula showed significant growth inhibition activity on human breast cancer MCF-7 cells and human cervical cancer HeLa cells,with IC50 values of 84.39 mg/L and 164.78 mg/L,respectively at 48 h.The best explants for callus induction were leaves and stem segments of C.pilosula.The optimal culture medium was MS+2,4-D 1mg/L+6-BA 0.5 mg/L+NAA 1mg/L.The optimum culture temperature and photoperiod of callus were at 25℃and 6 h/d light.The polysaccharide extracted from the callus pilosula of C.pilosula showed significant antitumor activity.

关 键 词：党参 组织培养 愈伤组织 多糖 抗肿瘤 

分 类 号：S567.53[农业科学—中草药栽培]