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Objective: The contents of five main components in Polygala tenuifolia from different habitats were determined, which provided certain data support and theoretical basis for the quality evaluation system of P. tenuifolia. Methods: The contents of polygalaxanthone III, 3,6’-disinapoyl sucrose, polygalacic acid, senegenin and tenuifolin of roots from different wild samples were determined by HPLC. Then SPSS 20.0 and SIMCA 11.5 were used for difference analysis, correlation analysis, hierarchical cluster analysis (HCA), and principal component analysis (PCA). Results: The content of the five main components in wild P. tenuifolia samples from different habitats was significantly different. The 20 samples were placed into two clusters (I, II) by HCA and PCA. Cluster I comprised three samples with higher content of 3,6’-disinapoyl sucrose, polygalaxanthone III, senegenin and tenuifolin from Weinan, Xianyang in Shannxi Province, and Xinjiang in Shanxi Province, whereas cluster II contained the other 17 samples. Conclusion: The results showed that the main components of P. enuifolia from Weinan, Xianyang in Shannxi Province and Xinjiang in Shanxi Province were significantly higher than other origins, and which provided a reference for the quality control, selection of excellent germplasm and cultivation bases of P. tenuifolia.
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Objective: To study the effects of salicylic acid (SA) and methyl jasmonate (MeJA) on growth, activity of related enzymes and chemical components in Polygala callus. Methods: The callus of Polygala was used as material. After 30 d of dark culture at different concentrations of SA (0, 4, 8, 12, 16, 20, 24, 28, 32 mg/L) and MeJA (0, 100, 200, 400, 600, 800, 1 000 μmol/L), the growth of callus, anti-oxidant enzyme activity, the content of MDA, total phenolic, total flavonoid, polygalaxanthone III, and 3,6’-disinapoyl sucrose were determined. Results: MeJA inhibited the growth of Polygala callus, and 12 mg/L SA promoted the growth of Polygala callus. SA and MeJA promoted the activity of SOD, CAT, POD, and MDA in the callus of Polygala. With the increase of SA and MeJA concentration, the activities of SOD, CAT and POD increased first and then decreased, and the content of MDA continued to rise. When the concentration of SA was 20 mg/L, the activities of CAT and SOD reached the maximum, which were 248.45 U/mg and 4451.06 U/mg, respectively. When the concentration of SA was 16 mg/L, the activity of POD reached the maximum, which was 7.22 U/mg. When the concentration of SA was 32 mg/L, the MDA content reached the maximum value of 25.09 nmol/mg. When the concentration of MeJA was 600 μmol/L, the activities of CAT, SOD, and POD reached the maximum, which were 273.30, 1451.06 and 15.27 U/mg, respectively. When the concentration of MeJA was 1000 μmol/L, the MDA content reached the maximum value of 27.10 nmol/mg. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenols. MeJA promoted the accumulation of total phenols and total flavonoids in Polygala callus. When MeJA concentration was 600 μmol/L, the content of total flavonoids was the highest. When the concentration of MeJA was 400 μmol/L, the total phenolic content was the highest. Both SA and MeJA promoted the accumulation of polygalaxanthone III and 3,6’-disinapoyl sucrose in Polygala callus. When the concentration of SA was 32 mg/L, the concentration of polygalaxanthone III, 3,6’-disinapoyl sucrose was the highest. When the concentration of MeJA was 1 000 μmol/L, the content of polygalaxanthone III, 3,6’-disinapoyl sucrose was the highest. Conclusion: MeJA had an inhibitory effect on the growth of Polygala callus, and 12 mg/L SA can promote this effect. SA and MeJA promoted the activity of SOD, POD, CAT, the content of MDA and the accumulation of polygalaxanthone III and 3,6’-disinapoyl sucrose in Polygala callus. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenolics. MeJA promoted the accumulation of total phenolics and total flavonoids in Polygala callus.
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Objective To investigate the effects of different drying methods on composition and content of five active constituents in root bark and root of Polgala tenuifoliaroot. Methods The contents of polygalaxanthone III, 3,6’-disinapoyl sucrose, polygalacic acid, senegenin, and tenuifolin in root bark and root from different drying samples were determined by HPLC. Then the data analysis was performed by ANOVA and TOPSIS methods. Results There is a difference in the order of drying methods for bark and root of P. tenuifoliaroot. For P. tenuifoliaroot root bark, the order of different drying methods was microwave drying > 60 ℃ hot-air drying > 50 ℃ hot-air drying > 70 ℃ hot-air drying > freeze-drying > 40 ℃ hot-air drying > shade drying > sun drying; For P. tenuifolia root, the different drying methods were sorted by microwave drying > 60 ℃ hot-air drying > shade drying > sun drying > 50 ℃ hot-air drying > 40 ℃ hot-air drying > 70 ℃ hot-air drying > freeze-drying. Conclusion Combined with the production practice, this study suggests that microwave drying and hot-air drying at 60 ℃ are suitable drying methods for P. tenuifoliaroot bark and root, providing a basis for the determination of drying methods for the origin processing of P. tenuifolia.
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Objective To study the growth physiology and components of Polygala tenuifolia under different light quality and intensity, and provide theoretical basis for the artificial cultivation, yield improvement, and increasing the target composition of P. tenuifolia. Methods Using P. tenuifolia shoots as the experimental material, four kinds of ights (white, yellow, red, and blue) as light quality, the light intensity was set at 100, 300, and 500 μmol/(m2∙s) to determinate the plant height, root length, leaf length, leaf width, malondialdehyde (MDA) content, antioxidant enzyme activity, and secondary metabolite content in P. tenuifolia after 30 d of illumination. Results Under the red light, P. tenuifolia had the highest plant height, the longest root length and leaf length, and the largest biomass. Under the yellow light, the MDA content was the lowest, and the MDA content was the highest under white light. The activities of SOD, POD, and CAT were significantly increased under red and blue light. The highest content of total flavonoids, total phenols, and 3,6'-disinapoyl sucrose under blue light. Different light qualities had no significant effect on the content of polygalaxanthone III. The biomass of plant height and the 3,6’-disinapoyl sucrose content were the highest under the light intensity of 300 μmol/(m2∙s). P. tenuifolia had the longest roots under the light intensity of 500 μmol/(m2∙s), and the content of MDA and the activities of POD, SOD, and CAT increased with the increase of light intensity. Conclusion Red light has the strongest promoting effect on the roots of P. tenuifolia. Red and blue light significantly increases the activity of POD, SOD, and CAT. Blue light promotes the accumulation of total flavonoids, total phenols, and 3,6’-disinapoyl sucrose. The light intensity of 500 μmol/(m2∙s) has the strongest promoting effect on the roots of P. tenuifolia, and the activities of POD, SOD, and CAT in leaves were higher. Different light qualities had no significant effect on the contents of total flavonoids, total phenols, and polygalaxanthone III. The light intensity of 300 μmol/(m2∙s) promotes the accumulation of 3,6’-disinapoyl sucrose.
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Objective To study and establish the HPLC fingerprint of Polgala tenuifolia for the identification and quality controlof P. tenuifolia. Methods Twenty batches of wild and cultivated P. tenuifolia collected from different regions in China were detected by HPLC. The Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012 A edition) was used to evaluate the similarity of the samples. The differences among the samples were identified by chemical pattern recognition methods including principal component analysis (PCA) and partial least squares discriminate analysis (PLS-DA). Results The common model of HPLC fingerprint of wild and cultivated P. tenuifolia was obtained, 24 common peaks were found in the chromatograph. The similarities between wild and cultivated P. tenuifolia fingerprints and control fingerprints in 20 batches from different regions were over 0.86, PCA results demonstrated obvious distinction between the wild and cultivated P. tenuifolia. The wild and cultivated P. tenuifolia was completely distinguished by PLS-DA. Twelve constituents, such as polygalactoneone III and 3,6’-disinapoyl sucrose were screened as biomarkers, representing the major differences between the two varieties. Conclusion The HPLC fingerprint combined with chemical pattern recognition can be used as an effective method for the quality control and identification of the different sources of P. tenuifolia, and provide a reference for the quality control and stoichiometric taxonomy of P. tenuifolia.
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Objective To establish the main component analysis method of commodity grade Polygala tenuifolia, and to explore the correlation between the grade character of P. tenuifolia and the main chemistry. Methods P. tenuifolia of four markets were determined by high performance liquid chromatography (HPLC) to analyze the different level of medicine and explore the correlation between commodity grade and composition for P. tenuifolia. Results The main components determination of P. tenuifolia in different market grades were not completely consistent with the grade of commodity. Conclusion There are certain limitations of the commodity grading standards for P. tenuifolia in the medicinal materials market. It is necessary to establish a new grade quality standard for accurately evaluating the quality of P. tenuifolia. This study also provides some reference for the comprehensive quantitative evaluation of P. tenuifolia.
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Aim To study the synergistic anti-depres-sion effect of 3 , 6-disinapoyl sucrose ( DISS ) and tenuifoliside A ( TFSA ) from Radix Polygalae and the preliminary mechanism . Methods Using the classical behavioral despair and depression model of mouse tail suspension test, 120 mice were divided into control group, positive group, DISS 5 mg·kg-1 group,DISS 10 mg·kg-1 group,TFSA 5 mg·kg-1 group,TFSA 10 mg· kg-1 group, DISS 5 mg · kg-1 +TFSA 5 mg · kg-1 group,DISS 5 mg·kg-1 +TFSA 10 mg·kg-1 group,DISS 10 mg·kg-1 +TFSA 5 mg·kg-1 group and DISS 10 mg · kg-1 +TFSA 10 mg · kg-1 group randomly. They were given intragastric injection for 7 days continuously, to observe the effect of DISS and TFSA monomer and its combination on the time of mouse tail suspension. Expression of BDNF in the hip-pocampus of mice was detected by immunohistochemis-try. The expressions of CREB, pCREB, CRTC1 and BDNF in the hippocampus of mice were detected by Western blot method. Results The administration of DISS and TFSA could shorten the immobility time of mice subjected to the tail. DISS ( 10 mg · kg-1 ) and TFSA( 10 mg · kg-1 ) group were significantly lower than single dose drug group(P<0. 05). DISS and TF-SA and the combination groups could increase the ex-pression of BDNF in hippocampus and cortex by immu-nohistochemistry(P <0. 05). At the same time, the contents of CREB, CRTC1, pCREB and BDNF protein in the hippocampus were increased by DISS and TF-SA, and the combination group was significantly higher than the single drug group ( P<0. 05 ) . Conclusion The administration of DISS and TFSA are used to acti-vate CREB transcription factor CRTC1 , and activate the phosphorylation of CREB in the hippocampus, and then increase the expression of BDNF in the hippocam-pus and plays a synergistic antidepressant effect.
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Objective: To investigate the glycolipids from Polygala sibirica var. megalopha and their structure-activity relationship of xanthine oxidase (XO) inhibitory activity. Methods: The 75% EtOH extract of P. sibirica var. megalopha was chromatographied with D101 macroporous resins column by water, 35% EtOH, 65% EtOH, and 95% EtOH, successively. Compared to other elution parts, the 65% EtOH elution part exhibited stronger XO inhibition. Further bioassay-guided separation led to the isolation of four known glycolipids, which demonstrated potent XO inhibition. Results: Four compounds were identified to be tenuifoliside A (1), 3,6'-disinapoyl sucrose (2), 3'-E-3,4,5-trimethoxycinnamoyl-6-benzoyl sucrose (3), and 3'-E-3,4,5-trimethoxycinnamoyl-4-benzoyl sucrose (4) through spectroscopic data analysis. All compounds were obtained from this plant for the first time. This enzyme inhibition was dose dependent and the IC50 values of compounds 1, 3, and 4 were 9.5, 10.2, and 7.7 μmol/L, respectively. Among them, compound 2 had significantly higher XO inhibitory activity than that of the control allopurinol (IC50=11.2 μmol/L). It was speculated that the styryl side chain linking with sugar base played an important role on the XO inhibitory activity. Conclusion: Glycolipides as a novel series of XO inhibitors, may be developed into a promising remedy for human gout.
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Objective: A rapid and specific ultra performance liquid chromatography (UPLC) method was established for simultaneous analysis on six compounds and fingerprint analysis on Polygalae Radix to evaluate the herb quality from different habitats in China. Methods: The UPLC method was carried out by gradient elution with acetonitrile-formic acid water (0.1%). The flow rate was 0.4 mL/min. The detection wavelength was at 320 nm. The fingerprint chromatograms and the contents of six compounds including sibiricose A5, sibiricose A6, sibiricaxanthone B, glomeratose A, polygalaxanthone III, and 3,6'-disinapoyl sucrose in 24 batches of Polygalae Radix were analyzed. The common peaks were identified by ultra performance liquid chromatography tandem with time-of-flight mass spectrometry with MSE data-acquistion mode (UPLC-Q-TOF-MSE). Results: There was a difference in contents of six compounds, especially for the content of 3,6'-disinapoyl sucrose and sibiricose A6. Thirty-seven peaks were selected as the common peaks, of which 33 peaks were identified, and the similarities of 24 batches were between 0.756 and 0.997. Based on the results of quantification and fingerprint analysis, a certain difference between samples from different habitats was further proven. Conclusion: The validated UPLC quantitative analysis and fingerprint methods are successfully used in the quality control of Polygalae Radix.
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OBJECTIVE: To carry out a comparison between the cortexes and the roots of Polygala tenuifolia, and to supply references for developing scientific processing method of P. tenuifolia. METHODS: A comprehensive comparison was made by comparing the chemical constitution and pharmacological activities of the cortexes and the roots of P. tenuifolia, i.e., multi-components determination and fingerprint analysis were used to analyze their chemical constituents, and the traditional pharmacological actions of P. tenuifolia, such as cough-relieving, sputum-removing and sedative effects were used to evaluate their efficacy. RESULTS: The chemical constitution of the cortexes, roots and heartwoods of P. tenuifolia was similar, but the contents of the various effective components in the cortexes were more than those in the roots and heartwoods. The pharmacological result revealed that there was no obvious difference between the cortexes and the roots of P. tenuifolia for their activities of cough-relieving, sputum-removing and sedative actions. The roots of P. tenuifolia showed a better tendency than the cortexes for cough-relieving and sputum-removing activities. CONCLUSION: In order to avoid the waste of crude drugs, and to save the cost during heartwood-discarding, the heartwoods of P. tenuifolia are suggested to be kept.