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ObjectiveTo explore the pharmacodynamic effects of total flavonoids of Paeonia suffruticosa flower （TFPFs） on rats with hyperuricemia and provide scientific data support for the research and development of therapeutic drugs for hyperuricemia. MethodThe hyperuricemia model was induced by adenine combined with ethambutol in rats. The rats were randomly divided into a blank control group， a model group， two positive control groups （allopurinol at 42 mg·kg-1 and Tongfengshu tablets at 600 mg·kg-1）， and high-， medium-， and low-dose TFPFs groups （260， 130， and 65 mg·kg-1）. The general conditions of rats were observed and recorded， and the body weight was recorded once every 5 days. The 24-hour urine volume， water intake， uric acid （UA）， and urinary protein of rats were determined after the last administration. The kidney index was calculated. The pathological changes in thymus and spleen tissues of rats were observed by hematoxylin-eosin （HE） staining. The serum activities of UA， creatinine （Cr）， blood urea nitrogen （BUN）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and total antioxidant capacity （T-AOC） of rats were determined. The xanthine oxidase （XOD） and adenosine deaminase （ADA） activities in the liver were detected. The content of uric acid transporter 1 （URAT1）， organic anion transporter 1 （OAT1）， and glucose transporter 9 （GLUT9） in the kidney was detected by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the results in the model group， TFPFs could improve the mental state of rats， increase the body weight（P<0.01）， promote UA excretion（P<0.01）， reduce the content of urinary protein（P<0.05）， relieve renal glomerular atrophy， renal tubular epithelial cell status， and urate crystal deposition in renal tubules， dwindle 24-hour urine volume， water intake， kidney index（P<0.05）， serum levels of UA， Cr， BUN， and MDA（P<0.05，P<0.01）， inhibit the activities of XOD（P<0.05） and ADA（P<0.05，P<0.01）in the liver， diminish the expression of GLUT9 in the renal homogenate（P<0.05）， and increase serum SOD and T-AOC activities as well as OAT1 expression（P<0.01） in the kidney. The pathological changes of thymus and spleen were improved. ConclusionTFPFs possess a protective effect on the kidney of rats with hyperuricemia， which is achieved by promoting uric acid excretion， inhibiting oxidation and the activity of key enzymes in uric acid synthesis， and regulating the expression of uric acid transporters.
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ObjectiveTo explore the pharmacodynamic effects of total flavonoids of Paeonia suffruticosa flower （TFPFs） on rats with hyperuricemia and provide scientific data support for the research and development of therapeutic drugs for hyperuricemia. MethodThe hyperuricemia model was induced by adenine combined with ethambutol in rats. The rats were randomly divided into a blank control group， a model group， two positive control groups （allopurinol at 42 mg·kg-1 and Tongfengshu tablets at 600 mg·kg-1）， and high-， medium-， and low-dose TFPFs groups （260， 130， and 65 mg·kg-1）. The general conditions of rats were observed and recorded， and the body weight was recorded once every 5 days. The 24-hour urine volume， water intake， uric acid （UA）， and urinary protein of rats were determined after the last administration. The kidney index was calculated. The pathological changes in thymus and spleen tissues of rats were observed by hematoxylin-eosin （HE） staining. The serum activities of UA， creatinine （Cr）， blood urea nitrogen （BUN）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and total antioxidant capacity （T-AOC） of rats were determined. The xanthine oxidase （XOD） and adenosine deaminase （ADA） activities in the liver were detected. The content of uric acid transporter 1 （URAT1）， organic anion transporter 1 （OAT1）， and glucose transporter 9 （GLUT9） in the kidney was detected by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the results in the model group， TFPFs could improve the mental state of rats， increase the body weight（P<0.01）， promote UA excretion（P<0.01）， reduce the content of urinary protein（P<0.05）， relieve renal glomerular atrophy， renal tubular epithelial cell status， and urate crystal deposition in renal tubules， dwindle 24-hour urine volume， water intake， kidney index（P<0.05）， serum levels of UA， Cr， BUN， and MDA（P<0.05，P<0.01）， inhibit the activities of XOD（P<0.05） and ADA（P<0.05，P<0.01）in the liver， diminish the expression of GLUT9 in the renal homogenate（P<0.05）， and increase serum SOD and T-AOC activities as well as OAT1 expression（P<0.01） in the kidney. The pathological changes of thymus and spleen were improved. ConclusionTFPFs possess a protective effect on the kidney of rats with hyperuricemia， which is achieved by promoting uric acid excretion， inhibiting oxidation and the activity of key enzymes in uric acid synthesis， and regulating the expression of uric acid transporters.
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Paeoniflorin is the main effective component of Paeoniae lactiflora and Paeonia suffruticosa. A large number of studies have proven that paeoniflorin has many pharmacological effects, such as anti-depression, anti-inflammation, analgesia, anti-tumor, liver protection, nerve protection, sedation and hypnosis, immunomodulation and so on. It has little toxic and side effects and has been highly concerned by people. At present, paeoniflorin is rarely used in clinical practice in the form of monomers. This article mainly refers to the related research literatures on paeoniflorin pharmacological action in recent three years, combs and summarizes the new progress of its pharmacological action research. The relationship between the drug effect and the prescription of traditional Chinese medicine was discussed, in order to provide a reference for further development and clinical application of paeoniflorin.
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In order to identify the endophytic fungus MD76 and isolate its secondary metabolite. METHODS: The identification of strain MD76 is based on morphological characteristics and sequence alignment analysis of 18s rDNA ITS, the secondary metabolites were isolated by chromatography technology; and their structures were elucidated by extensive spectroscopic analysis. RESULTS: The strain MD76 was identified as Phyllosticta capitalensis; and seven compounds were gained, including benzoic acid(1), 4-hydroxybenzoic acid(2), 4-hydroxybenzene propanoic acid(3), paeoniflorin(4), oxypaeoniflorin(5), gallic acid(6), apigenin(7). CONCLUSION: The endophytic fungus P. capitalensis was gained firstly from Paeonia suffruticosa and seven compounds were isolated from P. capitalensis for the first time.
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In this study, Paeonia suffruticosa roots and rhizospheric soil in five geographic regions which were harvested in October were utilized as experimental materials, then the diversity of endophytic and rhizospheric actinomycetes were investigated by High-throughput sequencing technique. The 1 754 operational taxonomic units (OTUs) were obtained from 129 954 high quality sequences, 1 311 OTUs were detected in rhizospheric actinomycetes and belonged to four classes, four orders, twenty-seven families and ninety-seven genera, thirty-three genera such as Ilumatobacter were found in the five regions rhizospheric soil while three genera such as Longispora were only detected in the Dao-di regions, the dominant genera were Mycobacterium, Nocardioides, Streptomyces. 443 OTUs were obtained in roots and distributed in three classes, three orders, twenty-four families and fifty genera, thirteen genera such as Cryptosporangium were found in the five regions roots while Planosporangium, Luteococcus were only detected in the Dao-di regions, the dominant genera were Nocardioides. Alpha diversity analysis showed that the Shannon and Chao1 index in rhizospheric actinomycetes in Bozhou, Tongling and Nanling region were higher than Heze and Luoyang. Based on principal co-ordinates analysis (PCoA) analysis, the rhizospheric actinomycetes formations were similar in Tongling and Nanling region, at the same in Tongling and Luoyang endophytic actinomycetes. According to heatmap analysis, Bozhou, Tongling and Nanling region rhizospheric actinomycetes showed a close similarity in actinomycetes community structures on phylogenetic analysis, while Tongling, Luoyang and Nanling endophytic actinomycetes showed the same. Our results not only suggested that the rich and diverse actinomycetes resources in P. suffruticosa roots and rhizospheric soil but also revealed rhizospheric actinomycetes in the Dao-di regions had high similarity.
Subject(s)
Actinobacteria , Paeonia , Phylogeny , Plant Roots , Soil MicrobiologyABSTRACT
Objective: To establish an HPLC digital fingerprint for flower petals of Danfeng (Paeonia ostii), and apply it in the determination of other six varieties of P. suffruticosa produced in Heze areas, while determining the contents of eight components in the flower petals. Methods: Nano-Micro C18 (250 mm × 4.6 mm, 5 μm) column was adopted and the gradient mobile phase consisted of acetonitrile (A)-0.1% formic acid (B) with a flow rate of 1.0 mL/min, the detective wavelength was 270 nm, and the column temperature was 25 ℃. Results: The chromatographic fingerprint similarity evaluation system for traditional Chinese medicines was used for analysis. The chromatographic fingerprint similarity of six batches of Danfeng samples was more than 0.993. There were altogether 14 common peaks, and eight of them were identified. A comparative analysis showed a similarity between samples of Danfeng and other six varieties produced in Heze. Meanwhile, the content of eight major chemical constituents, namely gallic acid, methyl gallate, kaempferol-3-O-β-D-glucopyranoside, apigenin-7-O-β-D-glucopyranoside, apigenin-7-O-β-D-neospheroside, dihydrokaempferol-7-O-β-D-glucopyranoside, paeoniflorin, and oxypaeoniflorin were determined in the range of 0.05-1.41, 1.01-4.41, 0.70-5.55, 4.60-18.30, 10.05-33.87, 0.09-1.76, 3.16-11.12, 0.85-2.02 mg/g. Conclusion: The combination of HPLC digital fingerprint and content determination could reflect its inherent quality in an all-round way, which provided scientific basis for the quality evaluation of P. suffruticosa.
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Objective: In order to understand the genetic relationship and genetic diversity of peony (Paeonia suffruticosa) in Hunan province, the DNA fingerprints of 47 peony germplasm materials of Hunan province were studied. Methods: Seven primers selected from 100 ISSR primers could obtain high polymorphism and reproducibility bands using ISSR markers. Results: Total 77 DNA bands were amplified including 67 polymarphic bands, counting for 87.01%. Genetic similarity analysis showed that Hunan peony germplasm resourses with high genetic diversity. Through the computer software analysis, peony variety tested the average effective number of alleles was 1.395 4, the average Nei's genetic diversity index is 0.248 0, the average Shannon's information index was 0.385 9; Between each type of the similarity coefficient of Jaccard was 0.532 5-0.961 0, with an average of 0.751 5, which was a sign of close genetic relationship between the materials. These 47 germplasm resourses were divided into two groups and three subgroups by UPGMA analysis, which could distinguish the varieties from different sources. Conclusion: It reveals from the molecular level of Hunan local peony varieties, genetic relationship, and genetic diversity so as to provides the theoretical basis for the varieties breeding worke of heat and humid resistance of peony.
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Endophytes were isolated and purified from the roots of medicinal plant Fengdan also known as Paeonia suffruticosa from Tongling region, Anhui province, China. Morphology and molecular biology methods were applied to indentify the endophyte strains. And methods of growth rate and filtering paper were also used for studying antibacterial/antifungal effects of the strains. As a result, 129 endophyte strains were isolated. Fifty-eight endophytic fungi strains were identified as 6 species in 4 genera and the dominant genus was Fusarium. Seventy-one endophytic bacteria strains were identified as 9 species in 3 genera and the dominant genus was Bacillus. Inhibitory diameter with endophytic fermenting liquid of Pseudomonas chlororaphis, F. nematophilum and B. megaterium from P. suffruticosa against Staphyloccocus aureus, Escherichia coli and B. subtilis reached 25.0,20.2,24.0 mm respectively. The inhibition rate of endophytic fermenting liquid from F. nematophilum against Penicillium sp. and Colletotrichum dematium reached 90.6% and 83.3%, respectively. The inhibition effect of P. chlororaphis against F. oxysporum f. sp. niveum and Mucor sp. was good and the antifungal rate reached 80.0% and 84.9%, respectively. P. suffruticosa in Tongling region contains abundant endophytes. P. chlororaphis and F. nematophilum are valuable species as starting strain about microbicide.
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Paeonia suffruticosa also named Moutan that cultivated in five geographic regions during different growth stages were chosen in this study. Biolog and 454 pyrosequencing technology were used to detect the whole microbial activity and fungal diversity for exploring the relationship between the geo-authentic features of the medicinal plant and the rhizosphere microorganism. The results suggest that the value of average well color development (AWCD) from the rhizosphere soil of P. suffruticosa in the five regions at the fo ur growth stage have an increasing tendency. 9 703 operational taxonomic unit (OTU) were obtained from 272 463 high quality sequences according to the similarity of 97% by the pyrosequencing. Fungi in five phyla, twenty-two classes, seventy orders, one hundred and thirty-nine families and two hundred and sixty-six genera were detected in the five regions excluding twelve percent to fifty-eight percent unidentified fungi. They were divided into four branches, i.e. Blastocladiales, Chytridiomycota, Dikarya and Glomeromycetes. Twenty-four genera such as Leptosphaeria were found in the five regions while six genera such as Curvularia were only detected in the geo-authentic regions. The dominant genera were Guehomyces, Exophiala and Fusarium in geo-authentic regions, whereas genus Leptosphaeria, Cryptococcus, Exophiala, Fusarium and Ceratobasidium in non-authentic regions. The results from principal co-ordinates analysis (PCoA) showed that the fungi formations were similar in Tongling and Nanling region at four different growth stages, and the same in Heze at the leaf bud and fruiting stage. According to heatmap analysis, Tongling and Nanling region showed a close similarity in fungal community structures on phylogenetic analysis, while Bozhou, Heze and Luoyang showed the same. In brief, the whole microbial activity was higher in geo-authentic regions than the non-authentic. Fungi in rhizosphere soil of the medicinal peony presented diversity and region specificity. We found not only the abundant new species in the five regions, but also the phylogenetic similarity in the geo-authentic regions.
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The seed of tree peony and herbaceous peony contained a variety of stilbenes which possess many pharmacological activities, such as antitumor, anti-inflammatory, allergy and neuraminidase inhibition. In order to develop and use peony seed resources, a simple and sensitive HPLC-DAD was developed for simultaneous determination of 10 stilbenes in peony samples, i.e.,suffruticosol A,suffruticosol B,suffruticosol C,trans-resveratrol,cis-ε-viniferin,trans-ε-viniferin,cis-suffruticosol D,cis-gnetin H,trans-suffruticosol D and trans-gnetin H. At the same time, the scavenging activity of DPPH free radicals was employed to evaluate their antioxidant effect. The results showed that the 10 stilbenes are mainly present in peony seed coat (total content of more than 16.7%) compared to peony seed kernel (total content less than 0.3%), and can be enriched in the extract of peony seed coat (total content of more than 75%) The extract of peony seed coat and 10 stilbenes exhibited significant antioxidant properties. This work provides a foundation for comprehensive utilization of the tree peony and herbaceous peony seed resources.
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This article was aimed to study the optimum extraction process of paeonol. The extraction yield of paeonol was taken as investigation index. And the best extraction process was screened by orthogonal experimental design. The results showed that the optimum condition of extraction process was to soak coarse powder of Paeonia suffruti-cosa into 15-fold water for 0.5 h, and then the distillation lasted for 2.5 h. The distillate was collected and cooled to room temperature. The crystallization lasted for 24 h at 4℃, and then filtered and dried for 48 h at room tempera-ture. It was concluded that the selected technology was stable, reasonable and feasible. The extraction yield of paeonol is over 80%.
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Objective: To detect gallic acid in Paeonia suffruticosa Andr. by high performance liquid chromatography-mass spectrometry. Methods: A method for determining gallic acid in Paeonia suffruticosa Andr. by HPLC-MS is described. The operating conditions were Diamonsil C18 column (5μm, 4.6×250 mm) at room temperature, methanol-0.05% phosphoric acid solution as mobile phase by step gradients elution at a flow of 0.8 ml/min and UV detection at 270 nm. Results:The content of gallic acid is 1.8 mg/g. The relative standard deviation of gallic acid is 1.7%. Conclusion:The content of gallic acid in Paeonia suffruticosa Andr. is high, which may be related to the inhibition of FAS.
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PURPOSE: The aim of the present study was to examine the effect of the root cortex of Paeonia suffruticosa, an anti-inflammatory folk medicine, on nitric oxide (NO) synthesis and the induction of inducible NO synthase (iNOS) gene expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. MATERIALS AND METHODS: We studied the effects of the RAW264.7 macrophage cell line of the extract from the root cortex of P. suffruticosa and NO with western blot and northern blot assays. RESULTS: We found that treatment of RAW264.7 macrophages with the ethylacetate fraction of the ethanolic extract of the root cortex of P. suffruticosa inhibits LPS-induced synthesis of NO in vitro. This effect is dose-dependent and appears to involve the suppression of iNOS gene expression, thereby reducing excessive NO synthesis in LPS-stimulated RAW264.7 macrophages. CONCLUSIONS: As NO is a pro-inflammatory molecule and iNOS inhibitors may have therapeutic potential, our findings may explain the anti-inflammatory property of the root cortex of P. suffruticosa. This root has been used in oriental folk medicine for the treatment of various inflammatory diseases with relatively little knowledge regarding its mechanism of anti-inflammatory action.
Subject(s)
Blotting, Northern , Blotting, Western , Cell Line , Ethanol , Gene Expression , Macrophages , Medicine, Traditional , Nitric Oxide , Nitric Oxide Synthase , Nitric Oxide Synthase Type II , PaeoniaABSTRACT
AIM: To probe into the method of field processed products of Paeonia suffruticosa Andr. and inspect its quality. METHODS: Through field processed investigation alcohol-macerated extracts and paenol content compared among the smoked, de-epidermis, drying and direct drying in the sun. RESULTS: Paeonia suffruticosa Andr. with the epidermis is better than others. CONCLUSION: The method of integrating field cutting crude drugs into pieces with processing of Paeonia suffruticosa Andr. has feasible standardization and industrial benefits.