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Objective To study the spectrum-effect relationship between GC-MS fingerprint of the volatile oil of Atractylodis Rhizoma and the dryness effect of Atractylodis Rhizoma, and clarify the main dryness components of Atractylodis Rhizoma. Methods The volatile oil of different batches of Atractylodis Rhizoma (S1-S9) were analyzed by GC-MS. Using the daily drinking amount of water, the viscosity of whole blood, and the content of aquaporin 2 (AQP2) in the kidney of rats as the indexes of dryness effect of Atractylodes Rhizome, the spectrum-effect relationship was analyzed by the combination of grey relational grade and orthogonal partial least square method. Results The dryness effect of Atractylodis Rhizoma was the combined action of multiple components, and the peaks of the major contribution to dryness effect were 31, 28, and 33, respectively. The corresponding components of the three peaks were β-eudesmol, (-)-aristolene, and bulnesol, which were identified by the spectral library retrieval analysis. Conclusion The main dryness effect components of Atractylodis Rhizoma are β-eudesmol, (-)-aristolene, and bulnesol, which provide a reference for investigating the material basis of dryness effect of Atractylodis Rhizoma.
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Objective: To explore the effect on the accumulation of medicinal compositions β-eudesmol, atractylon, atractylodin and key enzyme genes 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and farnesyl pyrophosphate synthase (FPPS) expression in biosynthesis of Atractylodes lancea under copper stress. Methods: Under copper stress, the expression of key enzyme genes HMGR and FPPS in A. lancea was determined by real-time fluorescence quantitative PCR; the content of three medicinal components in A. lancea were determined by HPLC; The correlation analysis was performed with SPSS, and DPS software for grey correlation analysis. Results: When the copper stress concentration was within 100 mg/kg, the expression of FPPS and the content of atractylon in the rhizomes of A. lancea increased slightly. However, when the copper concentration continued to increase, the expression levels of HMGR and FPPS and three medicinal components content of A. lancea showed a different degrees of downward trend. The expression levels of HMGR and FPPS were positively correlated with the content of β-eudesmol, atractylon, and atractylodin (P < 0.05) under copper stress. Grey relational analysis showed that the content of β-eudesmol and atractylon in the rhizomes was significantly correlated with the expression of HMGR and FPPS of A. lancea under copper stress. The expression of FPPS gene had the larger contribution on the composition of β-eudesmol and atractylon. However, the correlation between the content of atractylodin and the expression of these two key enzyme genes was relatively small. Conclusion: This study clarified the change regulation of two key enzyme gene expression and the content of three medicinal compositions, and revealed the relationship between β-eudesmol, atractylon and HMGR and FPPS, the key enzymes in terpene biosynthesis of A. lancea under copper stress. It contributed to the further study of the molecular regulation mechanism of the synthesis of medicinal constituents under copper stress and provided a theoretical basis for improving the quality of A. lancea.
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Objective: To study the content determination of atractylodin, atractylon, and β-eudesmol in Atractylodis Rhizoma by GC and characteristic spectrum in order to provide a scientific basis for the quality control. Methods: Using GC and Agilent HP-5 capillary column, taking nitrogen as carrier gas, FID as detector, temperature programming, split ratio, injection port temperature: 250℃, detector temperature: 250℃, column temperature: 130℃; The contents of atractylodin, atractylon and β-eudesmol of 25 samples between Chengde and purchased from other markets were determined by external standard method. The characteristic spectrum was set up and the similarity was analyzed by Estimating System of Similarity on the Chinese Medicine Fingerprint Chromatogram. Results: The determination method and characteristic spectrum by GC for atractylodin, atractylon, and β-eudesmol in Atractylodis Rhizoma were established. Nine characteristic peaks were identified; The linear range of β-eudesmol was 20.00 - 406.10 μg/mL (r = 0.999 9), and the average recovery was 100.75%, RSD = 1.17% (n = 6), and the limit of detection was 0.12 ng. The linear range of atractylon was 35.00 - 348.70 μg/mL (r = 0.999 5), and the average recovery was 99.84%, RSD = 1.29% (n = 6), and the limit of detection was 0.04 ng; The linear range of atractylodin was 16.46 - 329.30 μg/mL (r = 0.999 6), and the average recovery was 100.12%, RSD = 0.88% (n = 6), and the limit of detection was 0.06 ng. Conclusion: The concent determination and characteristic spectrum method of atractylodin, atractylon, and β-eudesmol established by this study are sensitive, simple, stability, which could make the determination result accurate and reliable.
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Objective: To establish a quantitative determination method for β-eudesmol in Cortex Magnoliae Officinalis by GC. Methods:β-Phenethanol was used as the internal standard substance;the column was a Zebron ZB-WAX capillary column ( 60 m × 320 μm,0. 5μm) with the column temperature of 200℃;the detector was FID and the vaporizer temperature was 250℃; the carrier gas was nitrogen with the flow rate of 1. 3 ml · min-1 and the split ratio was 4 ∶1. Results: The linear range of β-eudesmol was 0. 015 1-0. 271 2 mg·ml-1(r=0. 999 8);the average recovery was 99. 28%(RSD =1. 17%, n=6). Conclusion:The method is simple and accurate with good reproducibility, which can be used for the quality control of medicinal materials and decoction pieces of Cortex Magnoliae Officinalis.
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Throughout history, traditional herbal medicine has afforded a rich repository of remedies with diverse chemical structures and bioactivities against several health disorders. A common issue of herbal medicine is the limitation of information on their pharmacological activities and their active constituents. Traditionally, the use of herbal medicine has been based on empirical treatment and passed on from generation to generation with information available only in local journals. This prevents several herbal medicines from being developed to their full potential. The presentation will focus on research and development of Atractylodes lancea (Thunb) DC. (AL: family Compositae) as a potential chemotherapeutic for cholangiocarcinoma (CCA), the bile duct cancer commonly found in Southeast Asia. The dried rhizome of AL is a medicinal plant used in Chinese (“Cang Zhu”), Japan (“So-jutsu”) and Thai (“Khod-Kha-Mao”) traditional medicine for its various pharmacological properties including anticancer, anti-inflammation and antimicrobial activities, activities on central nervous, cardiovascular, and gastrointestinal systems. The major constituents in the essential oils from AL rhizome are β-eudesmol, hinesol and atractylon. Preliminary investigation has demonstrated its promising anti-CCA activity both <i>in vitro</i> and animal (Opisthorchis viverrini/dimethylnitrosamine-induced CCA in hamsters and CCA—xenografted nude mice) models with high selectivity index comparing with the standard drug, 5-fluorouracil. It also showed virtually no toxicity with only minimal CNS effects on locomotor activity at the maximum dose of 5,000 mg/kg body weight. Studies are underway to identify active constituent(s) which contribute to anti-CCA activity as well as its pharmacokinetic and pharmacodynamic properties. The main research interest of my research group is the discovery and development of traditional herbal medicine for the treatment of two important tropical diseases, cholangiocarcinoma and malaria. As the time is quite limited, I am going to give you the summary of the conceptual framework and highlight some important findings which will illustrate how different approaches have been used or applied for the discovery of the promising candidates for these two diseases.
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Objective: To investigate the effect of endophytic fungal elicitor on key enzyme activity, inducing pathway and mechanism involved in the secondary metabolites of Atractylodes lancea. Methods: NADPH oxidase, HMGR activities, the concentration of hydrogen peroxide (H2O2) and β-eudesmol were determined by the co-culture of endophytic fungal elicitor and A. lancea suspension cell. Results: NADPH oxidase activity was notably enhanced by Fusarium sp5 elicitor which could induce oxidative burst, significantly promote H2O2 accumulation, and activate HMGR in the sesquiterpenoids metabolic pathway. Compared with the control, the yield of β-eudesmol increased 257.6% and reached 66.59 μg/g. CAT and DPI could inhibit the HMGR activity and β-eudesmol biosynthesis in A. lancea cell induced by Fusarium sp5 elicitor. Exogenous H2O2 also induced HMGR and promoted the β-eudesmol biosynthesis. Conclusion: H2O2 is necessary to induce β-eudesmol synthetic signal molecule by activating the HMGR.