1.Functional characterization of CYP81C16 involved in the tanshinone biosynthetic pathway in Salvia miltiorrhiza.
Li REN ; Linglong LUO ; Zhimin HU ; Ying MA ; Jian WANG ; Yatian CHENG ; Baolong JIN ; Tong CHEN ; Jinfu TANG ; Guanghong CUI ; Juan GUO ; Luqi HUANG
Chinese Journal of Natural Medicines (English Ed.) 2023;21(12):938-949
Danshen, the dried roots and rhizomes of Salvia miltiorrhiza Bunge (S. miltiorrhiza), is widely used in the treatment of cardiovascular and cerebrovascular diseases. Tanshinones, the bioactive compounds from Danshen, exhibit a wide spectrum of pharmacological properties, suggesting their potential for future therapeutic applications. Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized, most of which belong to the CYP76 and CYP71 families. In this study, CYP81C16, a member of the CYP71 clan, was identified in S. miltiorrhiza. An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones, namely neocryptotanshinone, deoxyneocryptotanshinone, and danshenxinkuns A and B. SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%. Kinetic evaluations andin vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates. The overexpression of SmCYP81C16 promoted the accumulation of (iso)tanshinone in hairy root lines. The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones, either through microbial or plant metabolic engineering.
Humans
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Salvia miltiorrhiza/metabolism*
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Biosynthetic Pathways
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Quinones/metabolism*
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Plant Roots/metabolism*
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Gene Expression Regulation, Plant
2.Correlation between expression level of functional genes and tanshinones' accumulation in Salvia miltiorrhiza from different areas.
Beining LI ; Xiaoli ZHOU ; Luqi HUANG ; Xueyong WANG ; Chunsheng LIU
China Journal of Chinese Materia Medica 2011;36(24):3406-3409
OBJECTIVETo study the relationship between functional gene expression in Salvia miltiorrhiza from different producing areas and active principles, which might provide scientific basis for the gene regulation of tanshinones.
METHODThe quantitative determination of cryptotanshinone and tanshinone II A was carried out by using HPLC method, expression level of 3 functional genes of SmAACT, SmCMK and SmIPPI were investigated by real-time PCR method.
RESULTThe content of active principles together with expression level of SmAACT and SmCMK were higher in S. miltiorrhiza from genuine producing areas including Henan and Shanxi, but lower in samples from Beijing which was non-genuine producing area.
CONCLUSIONExpression level of SmAACT and SmCMK had close relationships involving tanshinones' accumulation, but the SmIPPI gene had not.
Diterpenes, Abietane ; metabolism ; Gene Expression ; Real-Time Polymerase Chain Reaction ; Salvia miltiorrhiza ; genetics ; metabolism
3.The effect of antioxidant on optimation of blood preservation.
Jing-Han LIU ; Wei HAN ; Feng-Lei LAI ; Yang YU ; Rui LI ; Xi-Lin OUYANG
Journal of Experimental Hematology 2003;11(6):646-649
In order to optimize the preservation of blood, 3 kinds of antioxidant were selected and each of them can be injected directly into vein, then the optimal dose of these antioxidants was chosen using statistical method; ISMC (injectio salvia miltiorrhizae composita), ginaton and the combination of ISMC and ginaton were added into blood as optimal dose, some references as ATP, EI and so on were observed during blood preservation. The results showed that all of the three kinds of antioxidants increased ATP, EI and decreased FHb during blood preservation. It is concluded that both of ISMC and ginaton can effectively optimize the preservation of blood and combination of ISMC and ginaton can produce additive effect.
Adenosine Triphosphate
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metabolism
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Antioxidants
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pharmacology
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Blood Preservation
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Erythrocytes
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drug effects
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physiology
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Humans
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Salvia miltiorrhiza
4.Effect of elicitors on induction and manipulation of secondary metabolic effective ingredients in Salvia miltiorrhiza.
Wenyuan LI ; Wei GAO ; Aijuan SHAO ; Yunfei HE ; Luqi HUANG
China Journal of Chinese Materia Medica 2011;36(3):258-262
The application of elicitors, which is currently the focus of research, has been considered as one of the most effective methods to improve the synthesis of secondary metabolites in medicinal plants. Biotic and abiotic elicitors can regulate the secondary metabolic pathways of effective ingredients in Salvia miltiorrhiza. This paper has introduced the research progress about the induction and the regulation mechanism of S. miltiorrhiza by elicitors.
Biosynthetic Pathways
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Salvia miltiorrhiza
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drug effects
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metabolism
5.Research progress of synthetic biology for tanshinones.
Wei GAO ; Tian-yuan HU ; Juan GUO ; Dong-mei LV ; Zhu-bo DAI ; Yong-jin ZHOU ; Lu-qi HUANG
China Journal of Chinese Materia Medica 2015;40(13):2486-2491
Synthetic biology research methods which design and build a new artificial biological systems (medicinal plants or microorganisms system) with specific physiological functions through clarifying and simulating the basic law of the biosynthesis of active components of traditional Chinese medicine, is considered to be a potential method to produce an abundant resources of bioactive components. Tanshinones is a kind of diterpene quinone compounds with important pharmacological activities from traditional Chinese medicine Salvia miltiorrhiza. This article systematically introduced the research progress of the synthetic biology of S. miltiorrhiza, in order to provide references for studies on other terpenoid bioactive components of traditional Chinese medicines, and give new research strategies for the sustainable development of traditional Chinese medicine resources.
Diterpenes, Abietane
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biosynthesis
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Medicine, Chinese Traditional
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Salvia miltiorrhiza
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metabolism
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Synthetic Biology
6.Effects of Ca2+ on salicylic-acid induced biosynthesis of salvianolic acid B in young seedlings of Salvia miltiorrhiza Bunge.
Rongrong CAO ; Bingyu XING ; Xiaolin DANG ; Yaqin YAO ; Liancheng LIU ; Juan'e DONG
Chinese Journal of Biotechnology 2013;29(12):1836-1846
In order to study the effects of Ca2+ in the biosynthesis of salvianolic acid B (Sal B) induced by salicylic acid (SA) in the young seedlings of Salvia miltiorrhiza, we used confocal laser scanning microscopy and high performance liquid chromatography to measure the change of relative fluorescence intensity of Ca2+ and the contents of Sal B induced by SA before and after the application of extracellular calcium channel inhibitors (VP and LaCl3), intracellular calcium channel inhibitor (LiCl), as well as intracellular calmodulin antagonist (TFP). SA could induce the calcium burst, and the Ca2+ peak could last to 2-3 min in the guard cells of S. miltiorrhiza, which prompted the biosynthesis of Sal B after the Ca2+ burst. Both Vp or LaCl3, and LiCl or TFP could inhibit the burst of Ca2+ and the biosynthesis of Sal B. The above results demonstrated that Ca2+ from the extracellular and the intracellular calcium store regulate the biosynthesis of Sal B elicited by salicylic acid in S. miltiorrhiz young seedlings.
Benzofurans
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metabolism
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Calcium
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metabolism
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Plant Leaves
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metabolism
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Salicylic Acid
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pharmacology
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Salvia miltiorrhiza
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metabolism
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Seedlings
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metabolism
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Signal Transduction
7.Discuss on model organism and model for geoherbs' study.
Luqi HUANG ; Zhubo DAI ; Dongmei LV ; Yuan YUAN
China Journal of Chinese Materia Medica 2009;34(9):1063-1066
The scientific essence of geoherbs bases on the attribute of matter. The phenotype of geoherbs from biological point view includes medicinal character, tissue structure, content and composition of the effective components and the therapeutic effect. The forming of geoherb is resulted from the interaction of genotype and environment. It can be expressed by the formula: phenotype = genotype + environmental modification. In order to prove this hypothesis, the authors discussed the model organism and model for geoherbs' study based on Salvia miltiorrhiza.
Environment
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Geology
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Models, Biological
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Phenotype
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Plant Extracts
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chemistry
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metabolism
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pharmacology
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therapeutic use
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Plants, Medicinal
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chemistry
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metabolism
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Salvia miltiorrhiza
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chemistry
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metabolism
8.Effects of plant-derived smoke water on accumulation of biomass and active substance of Salvia miltiorrhiza f. alba.
Jie ZHOU ; Lin ZOU ; Jia LI ; Xiao WANG ; Wei LIU ; Lei FANG ; Qiang LI ; Fang ZHANG ; Yong-Qing ZHANG
China Journal of Chinese Materia Medica 2014;39(13):2460-2463
To study the effect of plant-derived smoke water on the accumulation of biomass and active substance of Salvia miltiorrhiza f. alba, seedlings of S. miltiorrhiza were treated with different concentrations of smoke water (1:500, 1: 1 000, 1: 2 000). The fresh weight and dry weight of underground part, the number of split-root, maximum root diameter, average root diameter, average root length, the content of lipophilic components and water-soluble components were measured. Results showed that fresh weight and dry weight of underground part were respectively improved by 98.01%, 44.32% and 85.71%, 28.57% with significant difference by smoke water treatment with concentration of 1: 500 and 1: 1 000. Maximum root diameter and dry weight of underground part were respectively enhanced by 58.44% and 85.71% by smoke water with concentration of 1:500. The content of tanshinone I and tanshinone II(A) were improved by smoke water treatment, however there were no significantly difference on the content of cryptotanshinone and dihydrotan shinone. This study indicates that smoke water treatment could be used to improve the accumulation of biomass and active substance content of S. miltiorrhiza f. alba, which could provide new ideas for its green cultivating.
Agriculture
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methods
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Biomass
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Drugs, Chinese Herbal
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analysis
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metabolism
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Salvia miltiorrhiza
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chemistry
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growth & development
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metabolism
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Smoke
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analysis
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Water
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chemistry
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metabolism
9.Effects of compound salvia miltiorrhiza injection on aquaporin 3 expression in human amniotic epithelial cells.
Yi-Wen ZHANG ; Xiao-Yan MA ; Sheng-Di DING ; Ai-Lan XIE ; Xue-Qiong ZHU
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(1):51-55
OBJECTIVETo study the effects of Compound Salvia miltiorrhiza Injection (CSI) on aquaporin 3 (AQP3) expression in human amniotic epithelial cells (hAECs), and to explore its mechanisms for treating oligohydramnios.
METHODSThe hAECs selected from 8 human term pregnancies with oligohydramnios and no other complications (as the test group)and 8 human term pregnancies with normal amniotic fluid volume (as the control group) were primarily cultured. The mRNA and protein expressions of AQP3 in hAECs were detected using reverse transcription-polymerase chain reaction and Western blot with various concentrations of CSI (0.000, 0.001, 0.010, 0.020, 0.060, and 0.100 mg/mL, respectively) at different time points (0, 6, 12,24, and 48 h, respectively).
RESULTS(1) Compared with the control group, the AQP3 expression was down-regulated in the test group (P < 0.05). (2) The AQP3 expression in the two groups reached the peak when the concentration of CSI was 0.010 mg/mL, showing statistical difference when compared with other concentrations (P < 0.05). (3) The AQP3 expression reached the peak when 0.010 mg/mL CSI acted for 12 h, showing statistical difference when compared with other concentrations (P < 0.05). (4) The AQP3 expression was up-regulated in the two groups when 0.010 mg/mL CSI acted for 12 h. But the up-regulated AQP3 expression was more obvious in the test group than in the control group (P < 0.05).
CONCLUSIONSCSI could regulate the AQP3 expression in hAECs. CSI showed more obvious effects on the AQP3 expression in hAECs of oligohydramnios human term pregnancies.
Amnion ; cytology ; Aquaporin 3 ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; Female ; Humans ; Salvia miltiorrhiza
10.Characteristics of soil microbial variation during crop rotation period at cultivation area of Salvia miltiorrhiza in Zhongjiang of Sichuan province.
Guibing LIN ; Deguang WAN ; Xinjie YANG ; Kui ZHAO ; Yuxia ZHU ; Zhuyun YAN
China Journal of Chinese Materia Medica 2009;34(24):3184-3187
OBJECTIVETo study the characteristics of soil microbial variation during Salvia miltiorrhiza crop rotation.
METHODthe conventional cultivating microbial method was used to study the microbial number and communities structure and soil microbial biomass phosphorus (SMBP) was determined by chloroform vapor extraction method. The data was then analyzed by SPSS software.
RESULTWith the increase of the crop rotation years, the numbers of bacteria and actinomycetes in soil also, but the fungi and SMBP decreased.
CONCLUSIONMicrobial mechanism of crop rotation of the planting S. miltiorrhiza is the regulation of microbial number and bacteria physiological communities, the process rebuilds the soil ecological system balance. Microbial communities in soil need at lest 2 years to get to restore, after planting S. miltiorrhiza, which consisting with traditional planting experience.
Actinobacteria ; isolation & purification ; Bacteria ; isolation & purification ; Plants, Medicinal ; growth & development ; metabolism ; microbiology ; Salvia miltiorrhiza ; growth & development ; metabolism ; microbiology ; Soil Microbiology