Regulatory Effect of Polygala tenuifolia and Licorice-simmered P. tenuifolia on Learning and Memory， HPA Axis Function and Neurotransmitters in Rats with Heart-kidney Imbalance Insomnia / 中国实验方剂学杂志

Objective:To compare the regulatory effects of <italic>Polygala tenuifolia</italic> and licorice-simmered <italic>P. tenuifolia</italic> on learning and memory， hypothalamus-pituitary-adrenal axis （HPA axis） function and neurotransmitters in rats with heart-kidney imbalance insomnia. Method:The rat model of insomnia induced by multi-factor stimulation was established. After the model being made， the administration groups were given the extracts of <italic>P. tenuifolia</italic> and licorice-simmered <italic>P. tenuifolia</italic> by gavage （dose of 8 g·kg^-1·d^-1）， while the normal group and model group were given the same volume of normal saline for 7 days. Morris water maze was used to detect the changes of learning and memory ability of rats in each group. Enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of corticotropin-releasing hormone （CRH）， adrenocorticotropic hormone （ACTH） and cortisol （CORT） in serum of rats from each group. The contents of 5-hydroxytryptamine （5-HT）， dopamine （DA）， <italic>γ</italic>-aminobutyric acid （GABA） and glutamic acid （Glu） in the hypothalamus of rats were determined simultaneously by ultra performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS）. Result:Compared with the normal group， the spatial learning and memory ability of rats in the model group<italic> </italic>was decreased， the times and time of staying in target quadrant were significantly reduced （<italic>P<</italic>0.05， <italic>P<</italic>0.01）， the levels of CORT， CRH and ACTH in serum were significantly increased （<italic>P<</italic>0.01）， the contents of GABA， DA， 5-HT in hypothalamus tissue were significantly decreased （<italic>P<</italic>0.01）， and the content of Glu was significantly increased （<italic>P<</italic>0.01）. Compared with the model group， the spatial learning and memory ability of rats in the <italic>P. tenuifolia</italic> group and licorice-simmered <italic>P. tenuifolia </italic>group<italic> </italic>were increased， the times and time of staying in the target quadrant were significantly increased （<italic>P<</italic>0.05， <italic>P<</italic>0.01）， the levels of CORT， CRH and ACTH in serum were significantly decreased （<italic>P<</italic>0.05， <italic>P<</italic>0.01）， the contents of GABA， DA and 5-HT in hypothalamus tissue were significantly increased （<italic>P<</italic>0.05， <italic>P<</italic>0.01）， and the content of Glu was significantly decreased （<italic>P<</italic>0.05）. The recovery degree of each index in the licorice-simmered <italic>P. tenuifolia</italic> group was better than that in the <italic>P. tenuifolia</italic> group. Conclusion:Both <italic>P. tenuifolia</italic> and licorice-simmered <italic>P. tenuifolia</italic> can improve the learning and memory ability， improve the function of HPA axis， regulate the level of central neurotransmitters， and have the effect of calming the mind and improving the intelligence of rats with heart-kidney imbalance insomnia. The effect of licorice-simmered <italic>P. tenuifolia</italic> is better than that of <italic>P. tenuifolia.</italic>

Habitat Suitability Evaluation of Polygala tenuifolia Based on MaxEnt Model and ArcGIS / 中国实验方剂学杂志

Objective:Polygalae Radix is mainly produced in Northeast，North，Northwest and Central China and Sichuan province. There are obvious differences in quality of <italic>Polygala tenuifolia</italic> from different areas. <italic>P. tenuifolia</italic> cultivation areas are relatively concentrated. Currently，there are only a few studies about the suitability zoning of <italic>P. tenuifolia</italic> nationwide，in order to determine the suitable zoning of <italic>P. tenuifolia</italic> in China. This paper made a habitat suitability evaluation of <italic>P. tenuifolia</italic> nationwide based on analysis of ecological factors in distribution areas of sampling points. Method:MaxEnt model was used to select ecological factors that affected the distribution of <italic>P. tenuifolia</italic> and construct an evaluation model. ArcGIS software was used to evaluate the suitability of different habitats of<italic> P. tenuifolia</italic>. The suitable areas were classified into high，medium，low suitability areas and unsuitable area of <italic>P. tenuifolia</italic> nationwide. Result:The evaluation model was highly accurate，and concluded that <italic>P. tenuifolia'</italic>s suitability distribution area in China was 3.21×10⁶ km²，including 0.52×10⁶ km² of high suitability area，0.96×10⁶ km² of medium suitability area and 1.73×10⁶ km² of low suitability area. High suitability areas were mainly distributed in Shanxi province，Hebei province，Shandong province，Shaanxi province，Liaoning province and Henan province. The main ecological factors affecting the distribution of <italic>P. tenuifolia</italic> were annual average temperature change range，mean temperature of driest quarter，precipitation of wettest quarter，altitude，slope and topsoil （0-30 cm） calcium carbonate. Conclusion:The results of the study are consistent with the records in relevant literatures，and can provide a theoretical basis for protection and cultivation of Chinese medicine resource <italic>P. tenuifolia</italic>.

Content determination and quality analysis of five main components in Polygala tenuifolia from different habitats / 中草药

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.

Transcriptome analysis of Polygala tenuifolia seedlings induced by methyl jasmonate and key genes mining for triterpenoid biosynthetic pathway / 中草药

Objective: To obtain the transcriptome sequence database induced by methyl jasmonate (MeJA) and identify the genes related to the biosynthesis of triterpenoid saponin in Polygala tenuifolia. Methods: The seedlings grown for 30 d were respectively treated with sterile water, 50 μmol/L MeJA and 100 μmol/L MeJA for 24 h. The transcriptome data of seedlings of P. tenuifolia were obtained by Illunima HiseqTM 2000 150PE sequencing and de novo splicing of Unigene was realized by Trinity software. The GO classification, KOG functional annotation, metabolism of KEGG metabolic pathway, protein function annotation analysis, differential gene analysis and screening were completed based on BLAST. Results: A total of 52.19 Gb clean data were obtained after the transcriptome of P. tenuifolia being assembled by Trinity software, and 54 426 Unigenes were assembled with an average length of 1 604 bp. All Unigenes were annotated in the public databases NR, NT, KEGG, Swissprot, GO, and Pfam. Through differential analysis of genes responding to MeJA, a total of 3 390 differentially expressed genes (DEGs) were found, of which 1 287 were up-regulated and 2 103 were down-regulated. The response of DEGs showed that the total number and up-regulated number of P. tenuifolia seedlings treated by 100 μmol/L MeJA was the highest. KEGG enrichment analysis showed that differentially expressed genes were significantly enriched in metabolic pathways including phenylpropanoid biosynthesis, cysteine and methionine metabolism, starch and sucrose metabolism, carbon fixation in photosynthetic organisms and terpenoid backbone biosynthesis. Furthermore, a total of 59 Unigenes involved in anthraquinones biosynthesis were found according to the assignment of KEGG pathway. Expression analysis showed that AACT, HMGS, HMGR, MK, PMK, MPD, DXS, IDI, FPPS, SQS, SE and β-AS were up-regulated after being induced by MeJA. Conclusion: In this study, the transcriptome of P. tenuifolia seedlings treated with methyl jasmonate was analyzed, and candidate genes related to triterpenoid skeleton biosynthesis of P. tenuifolia were obtained. MeJA can induce the expression of genes related to triterpenoid skeleton synthesis, which provided a wealth of data resources for the molecular biology research and also laid the foundation for the analysis of the secondary metabolic pathways of triterpenoid saponins in P. tenuifolia.

Sedative and Hypnotic Effects and Transcriptome Analysis of Polygala tenuifolia in Aged Insomnia Rats / 中国结合医学杂志

OBJECTIVE@#To study the sedative and hypnotic effects and underlying mechanisms of Polygala tenuifolia (PT) on treating aged insomnia rats.@*METHODS@#Sixty Sprague-Dawley male rats were divided into 6 groups by a random number table, including control group, model group, diazepam group (0.92 mg/kg), as well as PT low-, medium- and high-dose groups (0.0875, 0.175, 0.35 g/kg, respectively), 10 rats in each group. Aged insomnia rat model was established with subcutaneous injection of D-galactose for 42 days and then intraperitoneal injection of para-chlorophenylalanine for 3 days. PT and diazepam were respectively given to aged insomnia rats by intragastric administration for 7 days after model establishment. Then the rats were investigated by body weight, Morris water maze test, pentobarbital test, enzyme-linked immunosorbent assay, and transcriptome sequencing.@*RESULTS@#Compared with the model group, PT increased the body weight, improved memory ability, and prolonged pentobarbital-induced sleep time of aged insomnia rats (P<0.01 or P<0.05). The medium dose of PT also increased the neurotransmitter levels of 5-hydroxytryptamine (5-HT) and gamma-aminobutyric acid (GABA), and decreased the level of Glu in the hippocampus of aged insomnia rats (P<0.05 or P<0.01). Twenty-four differentially expressed genes (DEGs) were overlapped among model group, medium-dose PT group, and diazepam group in transcriptome analysis. Fuom and Pcp2 were down-regulated by the treatment of medium-dose PT (P<0.01 or P<0.05). The metabolic pathways of PT were relatively less than diazepam (91 vs. 104).@*CONCLUSIONS@#The sedative and hypnotic effects of PT in aged insomnia rats might be related to neuro, metabolism pathways, especially through GABAergic signaling pathway. It provided more effective herb choice for the treatment of senile insomnia.

Allelopathic Effect of Water Extracts from Rhizosphere Soil of Three Medicinal Plants on Polygala tenuifolia Seeds and Their Seedlings / 中国实验方剂学杂志

Objective:To investigate the allelopathic effects of water extracts from rhizosphere soil of three medicinal plants Rehmannia glutinosa，Pinellia ternata and Isatis indigotica on seed germination and seedling growth of Polygala tenuifolia， screen the stubble varieties suitable for crop rotation with P. tenuifolia， and provide some scientific basis for continuous cropping obstacles of P. tenuifolia. Method:The bioassay method was used to study the effects of rhizosphere soil water extracts from three medicinal plants Rehmannia glutinosa，Pinellia ternata and Isatis indigotica at concentrations of 0.3，0.6，0.9 g·mL-1 on the germination of P. tenuifolia seed and seedling growth. Result:The rhizosphere soil water extracts of Rehmannia glutinosa and Pinellia ternata showed basically low-promotion and high-inhibition concentration effects on the final germination rate，germination potential，and germination index of P. tenuifolia seeds，while the water extract of Isatis indigotica showed significant allelopathic inhibition effect. All three rhizosphere soil water extracts showed significant allelopathic inhibition effects on the growth index of P. tenuifolia seedlings. Among them，the rhizosphere soil water extract of Rehmannia glutinosa showed lower inhibitory effect on the plant height and root length of P. tenuifolia seedlings than the other two water extracts. The photosynthetic pigment content，proline（Pro） content，and soluble sugar content of P. tenuifolia chinensis seedlings were the highest under 0.3 g·mL-1 soil water extract of Rehmannia glutinosa， with relatively higher content of soluble protein， and relatively lower content of hydrogen oxide（H2O2）. Under the treatment of 0.9 g·mL-1 soil water extract of Rehmannia glutinosa，P. tenuifolia seedlings had the highest peroxidase（POD） and superoxide dismutase（SOD） activities，low catalase（CAT） activity，and lowest content of malondialdehyde（MDA）. Conclusion:Based on the comprehensive analysis of the above experimental data and allelopathic effects，the water extract of rhizosphere of Rehmannia glutinosa can promote the germination of P. tenuifolia seeds to a certain extent，and lay the foundation for seedling resistance to biochemical stress. Therefore， Rehmannia glutinosa is more suitable for crop rotation with P. tenuifolia.

Effects of salicylic acid and methyl jasmonate on growth, activity of related enzymes and chemical components in Polygala tenuifolia Willd / 中草药

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.

Reference genes screening and spatio-temporal expression analyses of cytochrome P450 monoxoygenase genes in Polygala tenuifolia by qRT-PCR / 中草药

Objective: To provide a suitable expression analysis of Polygala tenuifolia in different growth years and tissues, so as to select the stable internal control genes. The spatial and temporal expression of four cytochrome P450 monooxygenase genes in the above-mentioned samples was analyzed. Methods: The soluble curve and Ct value of eight candidate internal reference genes including Tubulin 1, Tubulin 2, Elongation 1, Elongation 2, Actin 1, Actin 2 and cdc-42 in different growth years (1-3 years old) and tissues (root,stem, leaf, and flower) of P. tenuifolia were obtained by real-time quantitative PCR (qRT-PCR). The expression stability of candidate reference genes was assessed by geNorm and NormFinder. Then, the qRT-PCR technique was also used to identify the temporal and spatial expression of four P450 genes including CYP709B2, CYP71AP39, CYP88A85 and CYP714E38 in P. tenuifolia. Results: The geNorm results showed that the stable internal reference genes expressed in P. tenuifolia were Tubulin 2 and Elongation 1. The NormFinder results showed that the most stable and suitable internal reference gene for expression analysis was Elongation 1. The mRNA expression levels of CYP709B2 and CYP71AP39 genes in stems and leaves (1-3 years old) were higher than that in roots and flowers. The CYP88A85 and CYP714E38 genes had got a higher mRNA expression level in roots (1-3 years old). Conclusion: Elongation 1 is suitable as an ideal internal control gene for qRT-PCR analysis of P. tenuifolia. The expression trend of P450s in roots, stems, leaves and flowers of cultivated P. tenuifolia is inconsistent.

Effect of Different Hormone Ratios on Callus Induction and Flavone Accumulation in Polygala tenuifolia / 中国实验方剂学杂志

Objective:To study the effect of different hormone ratios on the callus induction of roots,stems and leaves of Polygala tenuifolia,and determine and analyze the amount of flavonoids in roots,stems and leaves of P. tenuifolia. Method:With MS as the basic medium and roots,stems and leaves of P. tenuifolia sterile seedings as explants,the effects of 2,4-D,NAA and 6-BA on callus induction and flavonoid accumulation in different parts of roots,stems and leaves of P. tenuifolia were determined by orthogonal test. Result:2,4-D,NAA and 6-BA had significant effects on the callus induction rate of roots,stems and leaves of P. tenuifolia. The optimal callus induction combination of leaves was MS+3.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.5 mg·L-1 6-BA,the optimal callus induction combination of stems was MS+1.0 mg·L-1 2,4-D+3.0 mg·L-1 NAA+1.5 mg·L-1 6-BA,the optimal callus induction combination for roots was MS+1.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.0 mg·L-1 6-BA. And 2,4-D,NAA and 6-BA had significant effects on flavonoid accumulation in the stem callus of P. tenuifolia,and MS+3.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+0.5 mg·L-1 6-BA was the best flavonoid accumulation combination.NAA,6-BA had significant effects on flavonoid accumulation in the leave callus of P. tenuifolia,while 2,4-D had no significant effect,and MS+3.0 mg·L-1 2,4-D+2.0 mg·L-1 NAA+1.0 mg·L-1 6-BA was the optimal flavonoid accumulation combination,the three hormones had no significant effect on the accumulation of flavonoids in the root callus of P. tenuifolia,and MS+2.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.5 mg·L-1 6-BA was the best flavonoid accumulation combination. Conclusion:Under the conditions,the callus induction rate of roots,stems and leaves of P. tenuifolia is 100%, especially, the callus of P. tenuifolia leaves was the optimal,which is followed by P. tenuifolia stems and P. tenuifolia roots. Under the conditions,the amount of flavonoids in roots,stems and leaves of P. tenuifolia reach 21.31,24.56,23.61 mg·g-1,respectively.

Effect of Hydrogen Peroxide (H2O2) on Content of Total Phenol Flavonoids and Activity of Related Enzymes in Polygala tenuifolia Callus / 中国实验方剂学杂志

Objective:To study the changes in total phenolic and flavonoid content and antioxidant enzyme activity of Polygala tenuifolia callus in MS medium with different concentrations of H2O2,in order to explore the physiological mechanism of Polygala tenuifolia callus in adapting to H2O2 environmental stress at the cellular level. Method:Five gradients of 0,5,10,15,20 mmol·L-1 were set for H2O2 concentration and added to MS medium, with P. tenuifolia callus as the experimental material. Total phenols,total flavonoids and antioxidant enzyme activities of callus were determined after 5,10,15,20,25 d of culture,respectively. Result:The contents of total phenols and flavonoids were the highest when the concentration of H2O2 was 5 mmol·L-1 for 15 d. The SOD activity was the highest when the callus was cultured for 5 d and the exogenous H2O2 concentration was 5 mmol·L-1. POD activity was the highest at 25 d and 5 mmol·L-1 concentration of exogenous H2O2.CAT activity was the highest at 25 d and 15 mmol·L-1 concentration of exogenous H2O2. Conclusion:P. tenuifolia callus has the ability to adapt to the environmental stress of H2O2 at a certain concentration. When it is subjected to the environmental stress of H2O2,P. tenuifolia callus can alleviate the damage by regulating its secondary metabolites and protecting enzyme system. It can significantly promote the content of total phenols and flavonoids in secondary metabolites at 5-10 mmol·L-1. SOD activity was significantly increased at 5 d and the concentration of exogenous H2O2 of 5 mmol·L-1. POD activity was significantly increased at 25 d and the concentration of exogenous H2O2 of 5 mmol·L-1. CAT activity was significantly increased at 25 d and concentration of exogenous H2O2 of 15 mmol·L-1.

Study on Quality Standard of Compound Platycodon grandiflorum Antitussive Tablets / 中国药房

OBJECTIVE： To establish the quality standard of Compound Platycodon grandiflorum antitussive tablets. METHODS： TLC was used to identify the P. grandiflorum， Polygala tenuifolia and Glycyrrhiza uralensis qualitatively in Compound P. grandiflorum antitussive tablets. HPLC-ELSD method was used to measure the content of platycodin D in Compound P. grandiflorum antitussive tablets. The determination was performed on Agilent C18 column with mobile phase consisted of acetonitrile-water （26 ∶ 74， V/V） at the flow rate of 1.0 mL/min. ELSD was used with drift tube temperature of 105 ℃， gas flow rate of 3.0 L/min and column temperature at 35 ℃. RESULTS： TLC chromatograms of P. grandiflorum， P. tenuifolia and G. uralensis had clear spots with good separation and no same spot from negative samples. The linear range of platycodin D was 0.421 9- 5.062 8 μg （r＝0.999 9）. The quantitative limit and detection limit were 0.364， 0.109 μg， respectively. RSDs of precision， stability， reproducibility and durability tests were all lower than 3.0％. The recovery rates were 87.32％-91.96％ （RSD＝1.73％，n＝6）. The platycodin D contents of 178 samples ranged from 0.004 to 0.73 mg/tablet. The content of platycodin D in 55 batches （30.9％） of samples was lower than the content limit （0.10 mg/tablet） proposed in this study. CONCLUSIONS： Established method is accurate and reliable， and can be used for the quality control of Compound P. grandiflorum antitussive tablets.

Metabolomics study on effects of compatibility of alcohol extracts of Magnolia officinalis and Polygala tenuifolia on urine metabolites in rats / 中国药理学通报

Aim: To study the effect of compatibility of Magnolia officinalis and Polygala tenuifolia on urine metabolites in rats, to explore its characteristic biomar-kers and analyze the related metabolic pathways, and to provide references for exploring the possible mechanisms of Magnolia officinalis relieving gastrointestinal motility disorders induced by Polygala tenuifolia. Methods: Forty male Sprague-Dawley rats were randomly divided into control group, Polygala tenuifolia 1.75 g · kg-1 group, Magnolia officinalis 3.50 g · kg-1 group, and the compatibility group (3.50 + 1.75) g · kg-1, continuously orally administered for three days. Urine was collected within 24 hours after the last administration and detected by UPLC-Q-TOF-MS in each group. The characteristic biomarkers and corresponding metabolic pathways were analyzed or screened by Progenesis QI V2.0, SIMCA-P 14.0, SPSS 20. 0, MetaboAnalyst 3. 0 and other softwares. Results: The metabolites in control group and drug-administered group could be clearly distinguished, and a total of 16 characteristic metabolites, 2-Aminobenzoic acid, D-Glucurono-6, 3-lactone and Homovanillin, were screened out. The main related pathways included Tyrosine metabolism, tryptophan metabolism, vitamin B6 metabolism and primary bile acid biosynthesis. Conclusions: The mechanism of Magnolia officinalis relieving gastrointestinal motility disorders induced by Polygala tenuifolia might be related with Tyrosine metabolism, tryptophan metabolism, primary bile acid biosynthesis and vitamin B6 metabolism.

Effects of Oligosaccharide Esters of Polygala tenuifolia on the Injury of Human Neuroblastoma SH-SY5Y Cells Induced by Aβ25-35 Fragment / 中国药学杂志

OBJECTIVE: To investigate the effects of oligosaccharide esters of Polygala tenuifolia on Aβ25-35 induced injury in SH-SY5Y cells and the relative mechanisms. METHODS: SH-SY5Y cells were cultured with Aβ25-35(25 μmol•L-1) for 24 h in the presence and absence of oligosaccharide esters component of Polygala tenuifolia. The cell viability was measured by MTT method; cell apoptosis was detected by flow cytometry; the LDH release, the SOD activity and MDA level were measured by biochemistry method; the Bcl-2 and Bax expression were determined by Western blotting. RESULTS: Oligosaccharide esters of Polygala tenuifolia could enhance the cell viability, decrease the apoptotic rate, the release of LDH and MDA level, and increase SOD activity. It also enhanced the expression of anti-apoptotic protein Bcl-2 and decreased the expression of apoptotic protein Bax. CONCLUSION: Oligosaccharide esters of Polygala tenuifolia have neuroprotective effects against the Aβ25-35 induced injury in SH-SY5Y cells, which is related to its anti-oxidation and anti-apoptosis.

Establishment for methods of fingerprint analysis and determination of index compounds on commercial Polygala tenuifolia by ultra performance liquid chromatography / 中草药

Objective: To establish a method of fingerprint analysis and determination of index compounds on commercial Polygala tenuifolia by ultra performance liquid chromatography, and to evaluate the quality of P. tenuifolia with different commodity specification grades. Methods: UPLC was used to establish different commodity specification grade of P. tenuifolia and determine the contents of ten indicative compounds including sibiricose A5, sibiricose A6, lancerin, sibiricoxanthone B, glomeratose A, 7-O-methyl mangiferin, polygalaxanthone III, 3,6′-disinapoyl sucrose, tenuifoliside A, and tenuifoliside C, and establish comprehensive quality evaluation function at the same time. Results: The methods for fingerprint analysis and compound determination were in agreement with methodological requirements. A total of 37 peaks were selected as the common peaks, of which 22 peaks were identified, and the similarities were between 0.927 and 0.971. There are differences at different degrees in the contents of ten indicative compounds. The comprehensive quality evaluation function showed that the overall quality of Datong was better than that of Zhongtong; The overall quality of wild P. tenuifolia was better than that of cultivated. Conclusion: The validated UPLC fingerprint analysis and compound determination methods were successfully used in the quality control of P. tenuifolia.

Network mechanism on effective constituents from Polygala Tenuifolia for anti-Alzheimer Disease / 中国药理学与毒理学杂志

OBJECTIVE To clarify out the network pharmacology mechanism of Polygala tenuifolia against Alzheimer disease(AD).METHODS Firstly,we collected the chemical constituents from Polyg-ala tenuifolia and key targets toward AD.Machine learning algorithms were applied to construct classifi-ers for predicting the effective constituents. Secondly, docking models were utilized for further evalua-tion.Finally,we built constituent-target,target-target network and target-biology pathway network.RE-SULTS 104 chemical constituents Polygala tenuifolia from were collected.Through prediction of blood-brain penetration and validation,36 chemical constituents were selected among 100 chemical constitu-ents,their action targets mainly focused on AChE,COX-2,TNF-α,insulin-degrading enzyme and APP. Their main structure types include Polygala saponins, Polygala glycosides, Polygala shrubby ketones, polygala xanthones and sterols,which acted on AchE,APP,M-TAU,GSK3β and 5HT1A with high fre-quency.Gene-Ontology and KEGG enrichment analysis showed that the main pathways of these con-stituents involve in neurotransmitter release,synaptic conduction and synaptic plasticity,apoptosis reg-ulation,phosphorylation pathway,Ca2+signaling pathway,and so on.CONCLUSION This study uncov-ered a network mechanism of Polygala tenuifolia against Alzheimer disease,which may provide impor-tant information for the further study and new drug development.

Screening and verification of effective ingredients of traditional Chinese medicine Q0409 in improving learning and memory ability of mice / 中国病理生理杂志

AIM:To screen and verify the effective ingredient of traditional Chines medicine (TCM) Q0409 in improving learning and memory ability of mice.METHODS:The mouse learning and memory impairment model was induced by intraperitoneal injection of scopolamine.The mice in each group were given the corresponding drug by gavage at the same time for 14 d in succession.Morris water maze test was used to measure the learning and memory ability of the mice, and then the hippocampal tissue homogenate was taken to determine the activity of acetylcholinesterase (AChE).The animals were divided into 8 groups according to L8(27) orthogonal table.The variance analysis and factorial analysis were used to analyze the pharmacological effects of seven kinds of single herbal in TCM Q0409 and determine the screening ingredients.The animals were divided into 6 groups according to the results of the preliminary screening results, and further testing and validation of TCM Q0409 screening ingredients were performed to get the final simplified ingredients.RESULTS:Three medicinal herbs of Polygalae, Panax ginseng and Acori graminei rhizome were screened by the orthogonal results of Morris water maze test and the activity of AChE in mouse hippocampal tissues.The simplified ingredients of TCM Q0409 were obtained through the variance results of Morris water maze test and the activity of AChE in mouse hippocampal tissues.CONCLUSION:Polygala and ginseng were eventually determined as simplified ingredients of TCM Q0409 and it was verified that they improve the learning and memory ability of the mice with learning and memory impairment.

Research Progress in Pharmacological Activities of Oligosaccharide Esters from Polygala tenuifolia / 中国药学杂志

Oligosaccharide esters have attracted more and more attention along with the study of the chemical constituents of Polygala tenuifolia. Modern pharmacological studies have shown that these compounds have many pharmacological activities, such as neuroprotection, antidepressant effect and improvement of learning ability and memory. This review summarized the research progress of pharmacological effects of oligosaccharide esters from Polygala tenuifolia to provide clues for further study of the neuroprotective effects of Polygala tenuifolia.

Quality research and analysis of Polygala tenuifolia with different commodity grade / 中草药

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.

Anti-light aging effect of Polygala tenuifolia saponins on the skin / 中国生化药物杂志

Objective To study the anti-aging effect of total saponins of Polygala tenuifolia (PTS). Methods The effects of total saponins of Polygala tenuifolia on oxygen free radicals were investigated by the methods of scavenging hydroxyl radical, superoxide radical and DPPH oxygen free radicals in vitro. The photoaging model of mice skin was established to observe the effect of total saponins of Polygala tenuifolia on mouse skin pathology. Results In the experiment of antioxidation in vitro, the total saponins of Polygala tenuiflora significantly scavenged hydroxyl free radicals (P<0.05, P<0.001) and superoxide radical (P<0.01) at 6.0 and 12.0mg/mL, DPPH free radicals (P<0.001). In the light aging experiment, the light aging model of mice skin was established successfully in group B (model + medium tea oil). Compared with group B, the density of the skin epidermis in D group (30 min before irradiation, 6mg/mL PTS) was thickened, the elastic fibers were arranged uniformly and tightly, the collagen fibers proliferated significantly and densely, and the inflammatory cells infiltration was not found. The treatment effect was most remarkable in group G (30min after irradiation, PTS 12mg/mL PTS), and the skin of the mice was thickened, the elastic fibers were evenly arranged, the collagen fibers had some hyperplasia, evenly distributed, and there was no infiltration of inflammatory cells, which had significant therapeutic effect. Conclusion The total saponins of Polygala tenuifoliahas significant antioxidant activity and anti - light aging effect.

Analysis of influencing factors of secondary metabolites contents in cultivated Polygala tenuifolia / 中国中药杂志

This work was launched to explore the effect of habitat and growth year on the secondary metabolites contents of cultivated Polygala tenuifolia. The samples of cultivated P. tenuifolia were analyzed by ultra-high performance liquid chromatography(UPLC)-quadrupole time-of-flight mass spectrometry(Q-TOF MS), and the obtained data were analyzed using multiple statistical analysis and cluster analysis. The results showed that compared with growth year, habitat is a main influencing factor which affected the secondary metabolites contents of P. tenuifolia. The contents of sucrose esters and oligosacchride multi-esters are greatly dependent on the habitat (the sample-AG with high levels of components of tenuifoliside B and tenuifoliside C, and the sample-FY with high levels of 3,6'-disinapoyl sucrose, tenuifoliose S, tenuifoliose L, and tenuifoliose V). There is no obvious effect of habitat and growth year on xanthone. The contents of triterpene saponins are greatly dependent on the growth year, and the content of parts of triterpene saponins increased as time goes on.The result indicated that the effect of habitat and growth year on different types of secondary metabolites is not completely equivalent. This study will contribute to the breeding of P. tenuifolia and amendment of current commodity criteria.