Identification of key genes and active anti-inflammatory ingredients in Panax medicinal plants by climate-regulated callus culture combined with gene-component-efficacy gray correlation analysis  

气候调控愈伤组织培养结合基因组分功效灰色关联分析鉴定人参属药用植物关键基因及抗炎活性成分

作　　者：Jiao Ai Yongshen Ren Long Cheng Xinqiao Liu Yanqiu Li Majing Jiao Yingzhou Wang Xin Deng Linlin Chen 艾娇;任永申;程龙;刘新桥;李艳秋;焦马静;汪迎洲;邓鑫;陈霖霖(中南民族大学药学院;湖北中医药大学药学院)

机构地区：[1]School of Pharmaceutical Science,South-Central University for Nationalities,Wuhan,Hubei,China [2]School of Pharmacy,Hubei University of Chinese Medicine,Wuhan,Hubei,China

出　　处：《Acupuncture and Herbal Medicine》2022年第4期261-273,共13页针灸和草药（英文）

基　　金：supported by National Natural Science Foundation of China grants(No.81773893);National Major Scientific and Technological Special Project for“Significant New Drugs Development”(No.2017ZX09301060001);Hubei Province Key R&D Programme(No.2020BED017 and 2020BGB004);Jiangxi Province“Thousand Talents Plan”of Scientific and Technological Innovation(No.JXSQ2019201105);Fundamental Research Funds for the Central Universities“South-Central University for Nationalities”(No.CZP20025 and No.CZP20047)

摘　　要：Objective:We aimed to establish a novel strategy for identifying key genes and active anti-inflammatory ingredients in Panax medicinal plants.Methods:First,fresh roots of 2-year-old Panax plants,including P.ginseng C.A.Mey.,P.quinquefolium L.,P.notoginseng(Burk.)F.H.Chen,P.japonicus C.A.Mey.,P.japonicus Mey.var.major(Burk.)C.Y.Wu et K.M.Feng,were selected as explants,and callus formation was induced under three experimental temperatures(17,24,and 30℃).Second,high-performance liquid chromatography-mass spectrometry was used to analyze the saponin content of the callus.Nitric oxide reduction efficacy was used for“component-efficacy”gray correlation analysis to find the active anti-inflammatory ingredients.Quantitative reverse-transcription polymerase chain reaction(qRT-PCR)was used to determine the inflammatory factors and verify the active ingredients’anti-inflammatory effects.Finally,qRT-PCR was used to detect the expression of key genes in the callus,and“gene-component”gray correlation analysis was used to examine the relationships between the regulatory pathway of the genes and the components.Results:Among the three experimental temperatures(17,24,and 30℃),the lowest temperature(17℃)is the most suitable for generating Panax callus.Lower-latitude native Panax notoginseng is more adaptable under high culture temperatures(24℃and 30℃)than other Panax plants.The ginsenoside contents of the callus of P.notoginseng and P.japonicus were the highest under similar climate conditions(17℃).Major anti-inflammatory components were G-Rh1,G-Rb1,G-Rg3,and G-Rh6/FloralGKa.CYP76A47 contributed to the accumulation of anti-inflammatory components.Conclusions:This study provides a strategy for the gene-component-efficacy correlational study of multi-component,multi-functional,and multi-purpose plants of the same genus.[目的]建立人参属药用植物关键基因和抗炎活性成分的鉴定新方法。[方法]首先,选择2年生人参、三七、西洋参、日本参、大叶人参的新鲜根诱导愈伤组织形成。其次,采用HPLC-MS分析愈伤组织中的皂苷,以一氧化氮还原为功效进行“成分功效”灰色关联分析,寻找抗炎活性成分,并采用qRT-PCR测定炎症因子,以验证活性成分的抗炎作用。最后,再次使用qRT-PCR检测愈伤组织中关键基因的表达,并使用“基因组分”灰色关联分析检查基因调控途径与组分之间的关系。[结果]在三个实验温度(17、24和30℃)中,低温(17℃)更适合人参愈伤组织的产生。与其他人参植物相比,低纬度天然三七在高温(24℃和30℃)下的适应性更强。相同气候条件下(17℃)三七和日本人参愈伤组织中人参皂苷含量最高。抗炎成分主要是包括G-Rh1、G-Rb1、G-Rg3和G-Rh6/GKa。CYP76A47对抗炎成分的积累贡献最大。[结论]本研究为同一属多组分、多功能和多用途植物的基因组分功效相关研究提供了策略。

关 键 词：Anti-inflammatory ingredient Gene-component-efficacy gray correlation analysis HPLC–MS PANAX qRT-PCR Callus culture 

分 类 号：R284[医药卫生—中药学]