北柴胡和狭叶柴胡中黄酮类成分及其关键酶基因表达的组织差异分析  

Analysis of tissues difference of flavonoids and key enzyme genes expression in Bupleurum chinense and B. scorzonerifolium

作　　者：杨林林 赵钰 韩梅[1] 杨利民[1] YANG Lin-lin;ZHAO Yu;HAN Mei;YANG Li-min(College of Chinese Medicinal Materials,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学中药材学院,吉林长春130118

出　　处：《中草药》2019年第1期188-194,共7页Chinese Traditional and Herbal Drugs

基　　金：现代农业产业技术体系建设专项资金资助(CARS-21)

摘　　要：目的研究北柴胡Bupleurum chinense、狭叶柴胡B. scorzonerifolium不同组织中黄酮类成分含量及其关键酶基因表达量的关系。方法以北柴胡及狭叶柴胡的根、茎、叶、果实为实验材料,采用HPLC法测定不同组织中黄酮类成分(芦丁、槲皮素、山柰素、异鼠李素)的含量,用紫外分光光度法测定总黄酮含量,用实时荧光定量PCR(q RT-PCR)法检测黄酮类成分关键酶基因IFS、F3H、DFR表达量,用SPSS软件进行相关性分析。结果北柴胡、狭叶柴胡地上部分(茎、叶、果实)黄酮类成分含量显著高于根,其中黄酮类成分主要为芦丁,芦丁在北柴胡叶中含量最高可达106.961 mg/g;北柴胡、狭叶柴胡中总黄酮分布有明显的组织差异,含量由高到低为叶≥果实>茎>根;北柴胡IFS、F3H、DFR基因地上部分表达量远高于根,其中IFS基因表达量与芦丁含量呈显著正相关(P<0.05),F3H基因和DFR基因的组织表达量呈显著正相关(P<0.05),而狭叶柴胡各组织IFS、F3H、DFR基因表达均处于较低水平。结论北柴胡、狭叶柴胡黄酮类成分组织含量与关键酶基因的组织表达有一定的一致性,关键酶基因的组织差异表达调控了黄酮类成分在不同组织中的合成与积累。Objective To investigate the relationship between the content of flavonoids and the key enzyme genes expression in different tissues of Bupleurum chinense and B. scorzonerifolium. Methods The roots, stems, leaves, and fruits of B. chinense and B. scorzonerifolium were used as test materials, determination of flavonoids(rutin, quercetin, kaempferol, and isorhamnetin) in different tissues by HPLC, determination of total flavonoids by UV spectrophotometry, the tissues expression of key enzyme genes(IFS, F3 H, and DFR) in flavonoids synthesis was determined by real-time quantitative PCR, correlation analysis was performed with SPSS. Results The content of flavonoids in the aerial parts(stems, leaves, and fruits) of B. chinense and B. scorzonerifolium was significantly higher than that in roots, the content of flavonoids was mainly rutin, and the content of rutin in the leaves of B. chinense leaves was up to 106.961 mg/g;The distribution of total flavonoids in B. chinense and B. scorzonerifolium was obviously different, the content was from high to low: leaves ≥ fruit > stem > root;The expression of B. chinense IFS, F3 H, and DFR gene in the aerial parts was much higher than that in roots, IFS gene was significantly positive correlated with rutin(P < 0.05), F3 H gene was significantly positive correlated with DFR gene(P < 0.05), but the expression of IFS, F3 H, and DFR gene in each tissues of B. scorzonerifolium was at lower level. Conclusion The content of flavonoids in different parts of B. chinense and B. scorzonerifolium was consistent with the expression of flavonoids synthesis key enzyme genes, the differential expression of key enzyme genes regulates the synthesis and accumulation of flavonoids in different tissues.

关 键 词：北柴胡 狭叶柴胡 芦丁 槲皮素 山柰素 异鼠李素 基因表达量 相关性 

分 类 号：R282.12[医药卫生—中药学]