PHARMACEUTICAL BIOLOGY
·
2017
·
IF 1.916
Xu-Xin Zhang,Yun-Feng Cao,Li-Xuan Wang,Xiao-Lin Yuan,Zhong-Ze Fang
Contents: Danshen is a popular herb employed to treat cardiovascular and cerebrovascular diseases worldwide. Danshen–drug interaction has not been well studied.Objective: The inhibitory effects of four major tanshinones (tanshinone I, tanshinone IIA, cryptotanshinone, and dihydrotanshinone I) on UDP-glucuronosyltransferases (UGTs) isoforms were determined to better understand the mechanism of danshen–prescription drugs interaction.Materials and methods: In vitro recombinant UGTs-catalyzed 4-methylumbelliferone (4-MU) glucuronidation reaction was employed. Tanshinones (100 μM) was used to perform the initial screening of inhibition capability. High-performance liquid chromatography (HPLC) was used to separate 4-MU and its glucuronide. In vitro–in vivo extrapolation (IV–IVE) was employed to predict in vivo inhibition situation.Results: Cryptotanshinone inhibited UGT1A7 and UGT1A9 with IC50 values of 1.91 ± 0.27 and 0.27 ± 0.03 μM, respectively. Dihydrotanshinone I inhibited UGT1A9-catalyzed 4-MU glucuronidation reaction with the IC50 value of 0.72 ± 0.04 μM. The inhibition of cryptotanshinone towards UGT1A7 and UGT1A9 was best fit to competitive inhibition type, and UGT1A9 was non-competitively inhibited by dihydrotanshinone I. Using in vitro inhibition kinetic parameters (Ki) and in vivo maximum plasma concentration (Cmax) of cryptotanshinone and dihydrotanshinone I, the change of area-under-the-concentration–time curve (AUC) was predicted to be 0.4–4.2%, 3.7–56.3%, and 0.6–6.4% induced by cryptotanshinone and dihydrotanshinone inhibition towards UGT1A7 and UGT1A9, respectively.Discussion and conclusion: The inhibitory effects of tanshinones towards important UGT isoforms were evaluated in the present study, which provide helpful information for exploring the mechanism of danshen–clinical drugs interaction.
研究领域:
药理学
·
中药学
·
代谢组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2016
·Vol. 6
·期 1
·1-11
·
IF 5.228
Shi Min,Zhou Wei,Zhang Jianlin,Huang Shengxiong,Wang Huizhong,Kai Guoyin
Jasmonic acid (JA) is an important plant hormone involved in regulation of many aspects of plant growth and development including secondary metabolism and JASMONATE ZIM-DOMAIN (JAZ) proteins are key components in JA signal processes. In this study, two new JAZ genes named SmJAZ3 and SmJAZ9 were cloned from S. miltiorrhiza hairy roots and characterized. Expression profiles under methyl jasmonate (MJ) treatment revealed that SmJAZ3 and SmJAZ9 were both MJ-responsive. Subcellular localization assay showed that SmJAZ3 was located in nucleus while SmJAZ9 was preferentially in nucleus. Expression of SmJAZ3 and SmJAZ9 in S. miltiorrhiza hairy roots differently affected the production of tanshinone. Over-expression of SmJAZ3 or SmJAZ9 in hairy roots produced lower level of tanshinone compared with the control, tanshinone production was as low as 0.077 mg/g DW in line SmJAZ3-3 and 0.266 mg/g DW in line SmJAZ9-22 . Whereas, down-regulation of SmJAZs enhanced tanshione production, the content of tanshinone increased to 2.48 fold in anti - SmJAZ3-3 line and 1.35-fold in anti-SmJAZ9-23 line. Our work indicated that SmJAZ3 and SmJAZ9 are involved in regulation of tanshinone biosynthesis and act as repressive transcriptional regulators in the JA signaling pathway, which paves the way to further dissect molecular mechanism in details in the future.
研究领域:
植物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
METABOLIC ENGINEERING
·
2022
·
IF 8.829
Min Shi,Ruiyan Zhu,Yi Zhang,Siwei Zhang,Tingyao Liu,Kunlun Li,Shucan Liu,Leran Wang,Yao Wang,Wei Zhou,Qiang Hua,Guoyin Kai
Phenolic acids and tanshinones are main bioactive compounds produced in Salvia miltiorrhiza widely used in treatment of cardiovascular diseases, which could be promoted by abscisic acid elicitation. However, the regulation mechanism remained to be elucidated. An ABA-inducible IIa WRKY transcription factor (TF) named SmWRKY34 exhibiting high homology with AtWRKY40 was isolated. SmWRKY34 exhibited a negative role on phenolic acids and tanshinones by directly regulating SmRAS and SmGGPPS . Moreover, ABA-responsive bZIP TF member named SmbZIP3 expressing significantly in SmWRKY34 transcriptome was screened. SmWRKY3 4 showed a negative regulatory role on SmbZIP3 . SmbZIP3 acted as a positive regulator in the biosynthesis of phenolic acids and tanshinones by targeting SmTAT and two tanshinone-promoting TFs SmERF128 and SmMYB9b . Taken together, we identify a new module WRKY34-bZIP3 involved in ABA signaling that manipulates phenolic acid and tanshinone accumulation, shedding new insights in metabolic engineering application in S. miltiorrhiza .
研究领域:
植物科学
·
分子生物学
·
药理学
·
转录组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF EXPERIMENTAL BOTANY
·
2020
·Vol. 71
·期 19
·5948-5962
·
IF 5.908
Deng Changping,Shi Min,Fu Rong,Zhang Yi,Wang Qiang,Zhou Yang,Wang Yao,Ma Xingyuan,Kai Guoyin
Phenolic acids and tanshinones are major bioactive ingredients in Salvia miltiorrhiza, which possess pharmacological activities with great market demand. However, transcriptional regulation of phenolic acid and tanshinone biosynthesis remains poorly understood. Here, a basic leucine zipper transcription factor (TF) named SmbZIP1 was screened from the abscisic acid (ABA)-induced transcriptome library. Overexpression of SmbZIP1 positively promoted phenolic acid biosynthesis by enhancing expression of biosynthetic genes such as cinnamate-4-hydroxylase (C4H1). Furthermore, biochemical experiments revealed that SmbZIP1 bound the G-Box-like1 element in the promoter of the C4H1 gene. Meanwhile, SmbZIP1 inhibited accumulation of tanshinones mainly by suppressing the expression of biosynthetic genes including geranylgeranyl diphosphate synthase (GGPPS) which was confirmed as a target gene by in vitro and in vivo experiments. In contrast, the phenolic acid content was reduced and tanshinone was enhanced in CRISPR/Cas9 [clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9]-mediated knockout lines. In addition, the previously reported positive regulator of tanshinone biosynthesis, SmERF1L1, was found to be inhibited in SmbZIP1 overexpression lines indicated by RNA sequencing, and was proven to be the target of SmbZIP1. In summary, this work uncovers a novel regulator and deepens our understanding of the transcriptional and regulatory mechanisms of phenolic acid and tanshinone biosynthesis, and also sheds new light on metabolic engineering in S. miltiorrhiza.
研究领域:
植物生物学
·
分子生物学
·
药理学
·
基因工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Chinese Journal of Natural Medicines
·
2017
·
IF 1.667
Xue-Jiao GUO,Xue-Jiao FAN,Bin QIAO,Zhi-Qiang GE
Danshensu [3-(3, 4-dihydroxyphenyl) lactic acid, DSS], one of the significant cardioprotective components, is extracted from the root of Salvia miltiorrhiza. In the present study, an ester prodrug of Danshensu (DSS), palmitoyl Danshensu (PDSS), was synthesized with the aim to improve its oral bioavailability and prolong its half-life. The in vitro experiments were carried out to evaluate the physicochemical properties and stability of PDSS. Although the solubility of PDSS in water was only 0.055 mg·mL −1 , its solubility in FaSSIF and FeSSIF reached 4.68 and 9.08 mg·mL −1 , respectively. Octanol-water partition coefficient (log P ) was increased from −2.48 of DSS to 1.90 of PDSS. PDSS was relatively stable in the aqueous solution in pH range from 5.6 to 7.4. Furthermore, the pharmacokinetics in rats was evaluated after oral administration of PDSS and DSS. AUC and t 1/2 of PDSS were enhanced up to 9.8-fold and 2.2-fold, respectively, compared to that of DSS. C max was 1.67 ± 0.11 μg·mL −1 for PDSS and 0.81 ± 0.06 μg·mL −1 for DSS. Thus, these results demonstrated that PDSS had much higher oral bioavailability and longer circulation time than its parent drug.
研究领域:
药理学
·
药物化学
✓ 已验证
DOI
高影响因子
PLANT CELL TISSUE AND ORGAN CULTURE
·
2021
·Vol. 147
·期 2
·389-404
·
IF 2.711
Shi Min,Hua Qiang,Kai Guoyin
Phenolic acids and tanshinones are important bioactive components with multiple pharmacological properties accumulated in traditional Chinese medicinal plant Salvia miltiorrhiza . Abscisic acid (ABA) is effective to enhance the content of the two compounds in S. miltiorrhiza , while the regulation mechanism remains unknown. Deep transcriptome profile of S. miltiorrhiza hairy roots treated with ABA was conducted. Totally, 42,710 unigenes with the average length of 1,157 bp were obtained from de novo assembly from about 2.81 billion produced clean reads, of which 27,641 unigenes (64.72%) were successfully annotated. Unigenes encoding key enzymes such as PAL, TAT, C4H, RAS, CYP98A14, GGPPS expressed significantly higher than that in control. Candidate laccases showing similar expression with RAS and CYP98A14 in phenolic acid biosynthesis and potential P450 unigenes involved in tanshinone biosynthesis genes were screened out. Furthermore, transcription factor (TF) profiles revealed that 1,250 TFs representing 35 families were identified, of which MYB, AP2/ERF, bHLH and bZIP were most abundant. The present study provided a comprehensive transcriptional expression profile affected by ABA, which will help underly the biosynthesis and regulation mechanism.
研究领域:
植物科学
·
分子生物学
·
药理学
·
转录组学
✓ 已验证
产品关联
DOI
高影响因子
FUNCTIONAL & INTEGRATIVE GENOMICS
·
2014
·Vol. 14
·期 3
·603-615
·
IF 2.691
Shi Min,Luo Xiuqin,Ju Guanhua,Yu Xiaohong,Hao Xiaolong,Huang Qiang,Xiao Jianbo,Cui Lijie,Kai Guoyin
Tanshinone is widely used for treatment of cardio-cerebrovascular diseases with increasing demand. Herein, key enzyme genes SmHMGR (3-hydroxy-3-methylglutaryl CoA reductase) and SmDXR (1-deoxy- d -xylulose 5-phosphate reductoisomerase) involved in the tanshinone biosynthetic pathway were introduced into Salvia miltiorrhiza ( Sm ) hairy roots to enhance tanshinone production. Over-expression of SmHMGR or SmDXR in hairy root lines can significantly enhance the yield of tanshinone. Transgenic hairy root lines co-expressing HMGR and DXR (HD lines) produced evidently higher levels of total tanshinone (TT) compared with the control and single gene transformed lines. The highest tanshinone production was observed in HD42 with the concentration of 3.25 mg g −1 DW. Furthermore, the transgenic hairy roots showed higher antioxidant activity than control. In addition, transgenic hairy root harboring HMGR and DXR (HD42) exhibited higher tanshinone content after elicitation by yeast extract and/or Ag + than before. Tanshinone can be significantly enhanced to 5.858, 6.716, and 4.426 mg g −1 DW by YE, Ag + , and YE-Ag + treatment compared with non-induced HD42, respectively. The content of cryptotanshinone and dihydrotanshinone was effectively elevated upon elicitor treatments, whereas there was no obvious promotion effect for the other two compounds tanshinone I and tanshinone IIA. Our results provide a useful strategy to improve tanshinone content as well as other natural active products by combination of genetic engineering with elicitors.
研究领域:
药理学
·
植物生物技术
·
代谢工程
✓ 已验证
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2017
·
IF 2.892
Yumin Fang,Zhuoni Hou,Xiaodan Zhang,Dongfeng Yang,Zongsuo Liang
Salvia miltiorrhiza Bunge ( S. miltiorrhiza ) and Salvia castanea Diels f. t omentosa Stib ( S. castanea ) are both important medicinal plants which have been widely used to treat cardiovascular diseases. Tanshinone and phenolic acid are active and specialized metabolites in the two species. However, the divergent mechanism of specialized metabolism in the two species is yet unknown. We found that S. castanea produced higher levels of cryptotanshinone, tanshinone IIA, caffeic acid, and rosmarinic acid, but lower levels of dihydrotanshinone I and salvianolic acid B than S. miltiorrhiza . Some key genes involved in specialized metabolism were potentially associated with the diverse metabolism between the two species. LH with 1 g l −1 contributed more to tanshinone accumulation in S. castanea than that in S. miltiorrhiza . Genes involved in tanshinone biosynthesis in S. castanea were also more sensitive to LH than those in S. miltiorrhiza . Knowledge about diverse specialized metabolism in the two species will help us understand mechanism of tanshinone and phenolic acid biosynthesis.
研究领域:
药理学
·
植物代谢
·
分子生物学
✓ 已验证
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHIC SCIENCE
·
2016
·Vol. 54
·期 8
·1435-1444
·
IF 1.32
Cao Jiliang,Wei Jinchao,Xiang Cheng,Zhang Mi,Li Baocai,Wan Jianbo,Su Huanxing,Li Peng
A simple and fast micellar electrokinetic chromatography (MEKC) method using ionic liquids as modifier was established for simultaneous determination of four hydrophobic tanshinones, including dihydrotanshinone I (1), cryptotanshinone (2), tanshinone I (3) and tanshinone IIA (4), in Danshen and related medicinal plants. In normal MEKC using sodium dodecyl sulfate (SDS) as surfactant and organic solvents as additives, the four tanshinones, especially cryptotanshinone and tanshinone I, could not be well separated. Fortunately, further addition of ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim]BF4) resulted in a baseline separation of these four analytes. After an optimization study, 10 mM borax–10 mM SDS–10 mM [bmim]BF4 containing 15% acetonitrile (v/v) at pH 9.6 was adopted as the running buffer to complete the separation within 16 min at the voltage of 25 kV, temperature of 25°C and detection wavelength of 254 nm. The relative standard deviations of migration time and peak area were in the range of 0.62–2.21 and 1.33–3.90%, respectively, indicating the good repeatability of the developed method. This method was extensively validated by evaluating the linearity (R2 ≥ 0.9992), limits of detection (0.75–1.11 μg mL−1), limits of quantification (2.26–3.32 μg mL−1) and recovery (96.11–103.74%). Under the optimum conditions, samples of Danshen and related medicinal plants were well analyzed with high separation efficiency.
研究领域:
分析化学
·
药物化学
·
药用植物
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Integrative Plant Biology
·
2020
·Vol. 62
·期 11
·1688-1702
·
IF 4.885
Changping Deng,Yao Wang,Fenfen Huang,Sunjie Lu,Limei Zhao,Xingyuan Ma,Guoyin Kai
MYB transcription factors play vital roles in plant growth and metabolism. The phytohormone methyl jasmonate (MeJA) promotes phenolic acid accumulation in the medicinal herb Salvia miltiorrhiza , but the regulatory mechanism is poorly understood. Here, we identified the MeJA-responsive R2R3-MYB transcription factor gene SmMYB2 from a transcriptome library produced from MeJA-treated S. miltiorrhiza hairy roots. SmMYB2 expression was tightly correlated with the expression of key salvianolic acid biosynthetic genes including CYP98A14 . SmMYB2 was highly expressed in the periderm of S. miltiorrhiza and SmMYB2 localized to the nucleus. Overexpressing SmMYB2 in S. miltiorrhiza hairy roots significantly increased the levels of salvianolic acids (including rosmarinic acid and salvianolic acid B) by upregulating salvianolic acid biosynthetic genes such as CYP98A14 . SmMYB2 binds to the MYB-binding motifs in the promoter of CYP98A14 , as confirmed by a dual-luciferase assay and electrophoretic mobility shift assays. Anthocyanin contents were significantly higher in SmMYB2- overexpressing hairy root lines than the control, primarily due to the increased expression of CHI , DFR , and ANS . These findings reveal the novel regulatory role of SmMYB2 in MeJA-mediated phenolic acid biosynthesis, providing a useful target gene for metabolic engineering and shedding light on the salvianolic acid regulatory network.
研究领域:
植物科学
·
分子生物学
·
天然药物学
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
Food & Function
·
2015
·Vol. 7
·期 1
·409-416
·
IF 2.791
Kaige Ji,Yudan Zhao,Tianhong Yu,Zhuoyi Wang,Hao Gong,Xin Yang,Yang Liu,Kun Huang
Parkinson's disease (PD) is one of the most common neurodegenerative diseases. Lewy bodies that are formed by the aggregated α-synuclein are a major pathological feature of PD. Salvia miltiorrhiza has been used as food and as a traditional medicine for centuries in China, with tanshinone I (TAN I) and tanshinone IIA (TAN IIA) as its major bioactive ingredients. Here, we investigated the effects of TAN I and TAN IIA on α-synuclein aggregation both in vitro and in a transgenic Caenorhabditis elegans PD model (NL5901). We demonstrated that TAN I and TAN IIA inhibited the aggregation of α-synuclein as demonstrated by the prolonged lag time and the reduced thioflavin-T fluorescence intensity; TAN I and TAN IIA also disaggregated preformed mature fibrils in vitro. Moreover, the presence of TAN I or TAN IIA affected the secondary structural transformation of α-synuclein from unstructured coils to β-sheets, and alleviated the membrane disruption caused by aggregated α-synuclein in vitro. Besides, the immuno-dot-blot assay indicated that TAN I and TAN IIA reduce the formation of oligomers and fibrils. We further found that TAN I and TAN IIA extended the life span of NL5901, a strain of transgenic C. elegans that expresses human α-synuclein, possibly by attenuating the aggregation of α-synuclein. Taken together, our results suggested that TAN I and TAN IIA may be explored further as potential candidates for the prevention and treatment of PD.
研究领域:
神经科学
·
药理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Advanced Research
·
2020
·
IF 6.992
Xiaolong Hao,Zhongqiang Pu,Gang Cao,Dawei You,Yang Zhou,Changping Deng,Min Shi,Shivraj Hariram Nile,Yao Wang,Wei Zhou,Guoyin Kai
Salvia miltiorrhiza Bunge is an herb rich in bioactive tanshinone and salvianolic acid compounds. It is primarily used as an effective medicine for treating cardiovascular and cerebrovascular diseases. Liposoluble tanshinones and water-soluble phenolic acids are a series of terpenoids and phenolic compounds, respectively. However, the regulation mechanism for the simultaneous promotion of tanshinone and salvianolic acid biosynthesis remains unclear. This study identified a R2R3-MYB subgroup 20 transcription factor (TF), SmMYB98 , which was predominantly expressed in S. miltiorrhiza lateral roots. The accumulation of major bioactive metabolites, tanshinones, and salvianolic acids, was improved in SmMYB98 overexpression (OE) hairy root lines, but reduced in SmMYB98 knockout (KO) lines. The qRT-PCR analysis revealed that the transcriptional expression levels of tanshinone and salvianolic acid biosynthesis genes were upregulated by SmMYB98-OE and downregulated by SmMYB98-KO . Dual-Luciferase (Dual-LUC) assays demonstrated that SmMYB98 significantly activated the transcription of SmGGPPS1 , SmPAL1 , and SmRAS1 . These results suggest that SmMYB98-OE can promote tanshinone and salvianolic acid production. The present findings illustrate the exploitation of R2R3-MYB in terpenoid and phenolic biosynthesis, as well as provide a feasible strategy for improving tanshinone and salvianolic acid contents by MYB proteins in S. miltiorrhiza .
研究领域:
植物生物技术
·
生药学
·
分子生物学
·
转录调控
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）