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Chinese Journal of Natural Medicines 2022年第8期20卷 614-626页

作者： DI Peng YAN Yan WANG Ping YAN Min WANG Ying-Ping huang lu-qi State Key Laboratory Breeding Base of Dao-Di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China Center for Post-doctoral Research China Academy of Chinese Medical SciencesBeijing 100700China State Local Joint Engineering Research Center of Ginseng Breeding and Application Jilin Agricultural UniversityChangchun 130118China

Panax quinquefolium is one of the most common medicinal plants worldwide.Ginsenosides are the major pharmaceutical components in P.quinquefolium.The biosynthesis of ginsenosides in different tissues of P.quinquefolium remained largely unknown.In the current study,an integrative method of transcriptome and metabolome analysis was used to elucidate the ginsenosides biosynthesis pathways in different tissues of P.quinquefolium.Herein,22 ginsenosides in roots,leaves,and flower buds showed uneven distribution patterns.A comprehensive P.quinquefolium transcriptome was generated through single molecular real-time(SMRT)and second-generation sequencing(NGS)technologies,which revealed the ginsenoside pathway genes and UDP-glycosyltransferases(UGT)family genes explicitly expressed in roots,leaves,and flower buds.The weighted gene co-expression network analysis(WGCNA)of ginsenoside biosynthesis genes,UGT genes and ginsenoside contents indicated that three UGT genes were positively correlated to pseudoginsenoside F11,notoginsenoside R1,notoginsenoside R2 and pseudoginsenoside RT5.These results provide insights into ginsenoside biosynthesis in different tissues ofP.quinquefolium.

关键词： Panax quinquefolium SMRT sequencing Ginsenoside biosynthesis UDP-glycosyltransferases

Molecular cloning and functional characterization of an isoflavone glucosyltransferase from Pueraria thomsonii

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Chinese Journal of Natural Medicines 2022年第2期20卷 133-138页

作者： DUAN Hai-Yan WANG Jian ZHA Liang-Ping PENG Hua-Sheng ZHAO Yu-Ping YUAN Yuan huang lu-qi School of Pharmacy Anhui University of Chinese MedicineHefei 230012China State Key Laboratory Breeding Base of Dao-di Herbs China Academy of Chinese Medical SciencesBeijing 100700China Research Unit of DAO-DI Herbs Chinese Academy of Medical SciencesBeijing 100700China

Pueraria thomsonii has long been used in traditional Chinese medicine.Isoflavonoids are the principle pharmacologically active components,which are primarily observed as glycosyl-conjugates and accumulate in P.thomsonii roots.However,the molecular mechanisms underlying the glycosylation processes in(iso)flavonoid biosynthesis have not been thoroughly elucidated.In the current study,an O-glucosyltransferase(PtUGT8)was identified in the medicinal plant P.thomsonii from RNA-seq database.Biochemical assays of the recombinant PtUGT8 showed that it was able to glycosylate chalcone(isoliquiritigenin)at the 4-OH position and glycosylate isoflavones(daidzein,formononetin,and genistein)at the 7-OH or 4′-OH position,exhibiting no enzyme activity to flavonones(liquiritigenin and narigenin)in vitro.The identification of PtUGT8 may provide a useful enzyme catalyst for efficient biotransformation of isoflavones and other natural products for food or pharmacological applications.

关键词： Flavonone Isoflavone Glucosyltransferase 4′-O-glucosylation 7-O-glucosylation

云苓延承与安苓兴起:清代地方志史料中茯苓产区及道地产区分析

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中国中药杂志 2024年第7期49卷 1981-1988页

作者：姜云璐彭华胜黄璐琦中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室北京100700 中医药文物研究国家文物局重点科研基地(中国中医科学院中药研究所) 北京100700

清代有丰富的地方志资料,为研究药材的生产提供了丰富的文献史料。该文整理了清代地方志中的茯苓相关内容,初步复原了清代茯苓在全国的分布情况。清代地方志中茯苓的分布涉及到23个省的318个清代县级区域。云南、安徽、湖南、浙江、福... 详细信息

清代有丰富的地方志资料,为研究药材的生产提供了丰富的文献史料。该文整理了清代地方志中的茯苓相关内容,初步复原了清代茯苓在全国的分布情况。清代地方志中茯苓的分布涉及到23个省的318个清代县级区域。云南、安徽、湖南、浙江、福建、江西、陕西、湖北等省份均有较多分布记录。清代地方志史料表明云南是茯苓的道地产区,与明清本草所述一致。安徽大别山区在清代茯苓品质优良,为新兴道地产区,习称“安苓”。安苓的发展,得益于从云南引进种植技术,并探索出特色的栽培技术,形成了包括种植、加工、售卖的完整茯苓产业链。清代地方志中茯苓史料非常丰富,不仅为道地产区的变迁提供了文献依据,也为福建、江西、湖南等重要的现代茯苓产区发展提供了历史脉络。除产区信息外,地方志还记录了各地茯苓品质、商品评价、栽培技法等内容,填补了本草古籍中的空白,为了解清代茯苓的产地与生产应用提供了翔实史料。

关键词：茯苓地方志清代道地药材产区

Genes functional identification and synthetic biology of natural products

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Chinese Journal of Natural Medicines 2020年第9期18卷 641-642页

作者： GAO Wei huang lu-qi School of Pharmaceutical Sciences Capital Medical UniversityBeijing 100069China State Key Laboratory Breeding Base of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China

Synthetic biology is one of the rapid developing scientific fields in recent years. Through synthetic biology,we have a better understanding of the natural synthesis process of natural products, which provides a favorable method to research the diversity of natural products, and also provides new tools for us to create new approach for producing natural products, we can synthesize compounds with different structures by artificial combination of different synthesis modules.

关键词： Genes functional identification and synthetic biology of natural products

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瑞香狼毒贝壳杉烷型二萜类合成酶功能表征

中国中药杂志 2024年第9期49卷 2410-2421页

作者：谭洪虎夏梦苏平黄璐琦南京中医药大学药学院江苏南京210023 中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室北京100700

ent-柯巴基焦磷酸合酶(ent-copalyl diphosphate synthase,CPS)与ent-贝壳杉烯合酶(ent-kaurene synthase,KS)的连续催化是植物以香叶基香叶基焦磷酸(geranylgeranyl pyrophosphate,GGPP)为底物开启赤霉素生物合成路径的关键步骤。该研究通过分析与挖掘瑞香狼毒的转录组数据,成功克隆得到赤霉素途径的2个关键二萜合酶基因SchCPS和SchKS。二者完整开放阅读框分别为2595、1701 bp,编码864、566个氨基酸残基,生物信息学分析预测其相对分子质量分别为97.9、64.6 kDa,理论等电点为5.61、6.12,均为亲水性蛋白。序列比对及系统发育树显示,SchCPS蛋白含有CPS基因家族保守的LHS、PNV及DxDD基序,归类于TPS-c亚家族;SchKS蛋白含有KS基因家族保守的DDxxD、NSE/DTE和PIx基序,归类于TPS-e亚家族。功能验证结果表明,SchCPS可催化GGPP质子化并环化成ent-CPP,SchKS则作用于ent-CPP去磷酸化并再次环化生成贝壳杉烯。该文首次从瑞香狼毒中克隆获得SchCPS与SchKS全长基因,并完成功能验证,不仅丰富了现有的CPS和KS基因库,也为瑞香狼毒中更多活性成分基因的克隆与生物合成途径解析奠定了基础。

关键词：瑞香狼毒贝壳衫烷二萜合酶功能鉴定生物信息学

洋地黄毒苷合成途径3β-羟基类固醇脱氢酶基因克隆及功能分析

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中国中药杂志 2024年第2期49卷 370-378页

作者：韩洋夏梦张逸风苏平黄璐琦中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室北京100700

洋地黄毒苷作为中药洋地黄中重要的次生代谢产物,是临床中最常用的强心药物之一。3β-羟基类固醇脱氢酶(3βHSD)是洋地黄毒苷生物合成途径酶,属于短链脱氢酶/还原酶(SDR)家族,起到氧化和异构前体化合物甾醇的作用,参与强心苷类化合物的... 详细信息

洋地黄毒苷作为中药洋地黄中重要的次生代谢产物,是临床中最常用的强心药物之一。3β-羟基类固醇脱氢酶(3βHSD)是洋地黄毒苷生物合成途径酶,属于短链脱氢酶/还原酶(SDR)家族,起到氧化和异构前体化合物甾醇的作用,参与强心苷类化合物的生物合成。该研究从洋地黄中首次克隆得到2个3βHSD基因,实验结果显示:Dp3βHSD1开放阅读框为780 bp,编码259个氨基酸;Dp3βHSD2开放阅读框为774 bp,编码257个氨基酸。Dp3βHSD1/2均具有TGxxxA/GxG辅因子结合位点和YxxxK催化位点基序,含有SDR酶的活性位点和NAD(P)结合位点。通过体外酶促试验证实Dp3βHSD1/2均能催化孕烯醇酮生成洋地黄毒苷前体化合物黄体酮,且Dp3βHSD1较Dp3βHSD2具有更强的催化能力。基因表达水平分析表明Dp3βHSD1基因在叶中的表达水平显著高于Dp3βHSD2,而洋地黄毒苷在叶中特异性富集,基于以上结果推测洋地黄毒苷生物合成途径中孕烯醇酮脱氢生成黄体酮过程主要由Dp3βHSD1催化完成。该研究成功解析了功能性3β-羟基类固醇脱氢酶,为进一步解析洋地黄中强心苷类化合物的生物合成途径奠定基础。

关键词：洋地黄洋地黄毒苷 3β-羟基类固醇脱氢酶功能鉴定

Factors Defining the Development of Severe Illness in Patients with COVID-19:A Retrospective Study

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Biomedical and Environmental Sciences 2021年第12期34卷 984-991页

作者： XIONG Yi Bai TIAN Ya Xin MA Yan YANG Wei LIU Bin RUAN Lian Guo lu Cheng huang lu qi Institute of Basic Research in Clinical Medicine China Academy of Chinese Medical SciencesBeijing 100700China Department of Infectious Diseases JinYinTan HospitalWuhan 430024HubeiChina National Resource Center for Chinese Materia Medica China Academy of Chinese Medical SciencesBeijing 100700China

Objective Early triage of patients with coronavirus disease 2019(COVID-19) is pivotal in managing the disease. However, studies on the clinical risk score system of the risk factors for the development of severe disease are limited. Hence, we conducted a clinical risk score system for severe illness, which might optimize appropriate treatment strategies.Methods We conducted a retrospective, single-center study at the JinYinTan Hospital from January 24,2020 to March 31, 2020. We evaluated the demographic, clinical, and laboratory data and performed a10-fold cross-validation to split the data into a training set and validation set. We then screened the prognostic factors for severe illness using the least absolute shrinkage and selection operator(LASSO)and logistic regression, and finally conducted a risk score to estimate the probability of severe illness in the training set. Data from the validation set were used to validate the score.Results A total of 295 patients were included. From 49 potential risk factors, 3 variables were measured as the risk score: neutrophil to lymphocyte ratio(OR, 1.27;95% CI, 1.15–1.39), albumin(OR,0.76;95% CI, 0.70–0.83), and chest computed tomography abnormalities(OR, 2.01;95% CI, 1.41–2.86)and the AUC of the validation cohort was 0.822(95% CI, 0.7667–0.8776).Conclusion This report may help define the potential of developing severe illness in patients with COVID-19 at an early stage, which might be related to the neutrophil to lymphocyte ratio, albumin, and chest computed tomography abnormalities.

关键词： COVID-19 Risk factors Severe illness Nomogram

Molecular cloning and functional characterization of multiple ApOSCs from Andrographis paniculata

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Chinese Journal of Natural Medicines 2020年第9期18卷 659-665页

作者： WANG Jian LIN Hui-Xin ZHAO Huan GUO Juan SU Ping YANG Jian WU Xiao-Yi huang lu-qi GAO Wei School of Traditional Chinese Medicine Shenyang Pharmaceutical UniversityShenyang 100016China State Key Laboratory Breeding Base of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China School of Traditional Chinese Medicine Capital Medical UniversityBeijing 100069China Geneis(Beijing)Co..Ltd Beiing 100102Chind.

Triterpenoids have been described in Andrographis paniculata. Oleanolic acid exhibits high biological activity and is widely used in the clinic, and β-sitosterol not only has good biological activity but also plays an important physiological role in plants.However, analysis of the biosynthetic pathway of triterpenoids in Andrographis paniculata has not been reported. Here, we provide the first report of the isolation and identification of nine 2, 3-oxidosqualene cyclases(ApOSC3 to ApOSC11) from A. paniculata. The results showed that ApOSC4 represented a monofunctional synthase that could convert 2, 3-oxidosqualene to β-amyrin. ApOSC5 as a bifunctional 2, 3-oxidosqualene cyclases, could transfer 2, 3-oxidosqualene to β-amyrin and α-amyrin. ApOSC6 to ApOSC8 composed the multifunctional 2, 3-oxidosqualene cyclases that could convert 2, 3-oxidosqualene to β-amyrin, α-amyrin and one or two undetermined triterpenoids. This study provides a better understanding of the biosynthetic pathway of triterpenoids in A. paniculata, and the discovery of multifunctional 2, 3-oxidosqualene cyclases ApOSC5 to ApOSC8 of the facilitates knowledge of the compounds diversity in A. paniculata.

关键词： Triterpenoids 2 3-Oxidosqualene cyclases A.paniculata β-Amyrin α-Amyrin

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紫丁香中1个新的倍半萜类化合物

中国中药杂志 2024年第9期49卷 2461-2467页

作者：邰巴达拉胡孙天霞代那音台虞慕瑶刘琪黄璐琦郑汉内蒙古民族大学蒙医药学院国家药品监督管理局中药(蒙药)质量控制重点实验室内蒙古通辽028000 中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室北京100700 长春中医药大学吉林省人参科学研究院吉林长春130117

该研究选定紫丁香结香部位乙醇提取物,通过硅胶柱色谱、薄层板制备以及制备液相色谱等进行系统分离与纯化,并结合紫外分析(UV)、红外分析(IR)、核磁共振(NMR)、高分辨质谱(HR-ESI-MS)、X射线衍射等波谱技术及文献理化数据比较等方法进... 详细信息

该研究选定紫丁香结香部位乙醇提取物,通过硅胶柱色谱、薄层板制备以及制备液相色谱等进行系统分离与纯化,并结合紫外分析(UV)、红外分析(IR)、核磁共振(NMR)、高分辨质谱(HR-ESI-MS)、X射线衍射等波谱技术及文献理化数据比较等方法进行结构鉴定,共鉴定出10个化合物,分别为紫丁香倍半萜D(1)、(-)-T-muurolol(2)、紫丁香倍半萜E~G(3~5)、14-noreudesma-3-hydroxy-3-en-2,9-dione(6)、1-isopropyl-2,7-dimethylnaphthalene(7)、异水菖蒲二醇(8)、α-二去氢菖蒲烯(9)、cadin-4-en-1-β-ol(10)。其中化合物1为未见报道的倍半萜类新化合物,其余9个化合物均为首次从紫丁香中分离得到。化合物1对脂多糖(LPS)诱导的RAW264.7细胞炎性损伤模型具有显著的保护作用。

关键词：紫丁香倍半萜抗炎活性

Differential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii

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Chinese Journal of Natural Medicines 2019年第8期17卷 575-584页

作者： TONG Yu-Ru ZHANG Yi-Feng ZHAO Yu-Jun HU Tian-Yuan WANG Jia-Dian huang lu-qi GAO Wei School of Pharmacy Traditional Chinese Materia Medica Shenyang Pharmaceutical University Shenyang 110016 China National Resource Center for Chinese Materia Medica China Academy of Chinese Medical Sciences Beijing 100700 China School of Pharmaceutical Science Capital Medical University Beijing 100069 China School of Traditional Chinese Medicine Capital Medical University Beijing 100069 China Advanced Innovation Center for Human Brain Protection Capital Medical University Beijing 100069 China

3-Hydroxy-3-methylglutaryl-CoA synthase(HMGS) is the first committed enzyme in the MVA pathway and involved in the biosynthesis of terpenes in Tripterygium wilfordii. The full-length cDNA and a 515 bp RNAi target fragment of TwHMGS were ligated into the p H7 WG2 D and p K7 GWIWG2 D vectors to respectively overexpress and silence, Tw HMGS was overexpressed and silenced in T. wilfordii suspension cells using biolistic-gun mediated transformation, which resulted in 2-fold increase and a drop to70% in the expression level compared to cells with empty vector controls. During Tw HMGS overexpression, the expression of TwHMGR, TwDXR and TwTPS7 v2 was significantly upregulated to the control. In the RNAi group, the expression of Tw HMGR,TwDXS, TwDXR and TwMCT visibly displayed downregulation to the control. The cells with TwHMGS overexpressed produced twice higher than the control value. These results proved that differential expression of Tw HMGS determined the production of triptolide in T.wilfordii and laterally caused different trends of relative gene expression in the terpene biosynthetic pathway. Finally, the substrate acetyl-Co A was docked into the active site of TwHMGS, suggesting the key residues including His247, Lys256 and Arg296 undergo electrostatic or H-bond interactions with acetyl-CoA.

关键词： Overexpression RNAi HMGS Triptolide Acetyl-CoA