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OBJECTIVE To establish comprehensive quality evaluation method based on multi-index components combined with multivariate statistical analysis， and to comprehensively evaluate the quality of Periploca forrestii. METHODS Taking 11 batches of P. forrestii medicinal materials from different areas in Guizhou as samples， the contents of neochlorogenic acid， cryptochlorogenic acid， chlorogenic acid， procyanidin A2， isochlorogenic acid A and isochlorogenic acid C were determined by HPLC. Clustering heat map analysis， grey correlation analysis（GRA） and technique for order preference by similarity to ideal solution（TOPSIS） were used to evaluate the quality of P. forrestii. RESULTS The results of methodological investigation of content determination were in accordance with the relevant regulations， and the linear relationship and accuracy of each component were good in their respective sampling range. The contents of chlorogenic acid， cryptochlorogenic acid， neochlorogenic acid， procyanidin A2， isochlorogenic acid A and isochlorogenic acid C in 11 batches of samples were 3.650-7.302， 0.888-2.575， 1.371- 2.386， 0.947-1.469， 0.084-0.169 and 0.725-1.067 mg/g， respectively. The content of each component was significantly different， with the highest content of chlorogenic acid and the lowest content of isochlorogenic acid A. The comprehensive results of cluster heat map， GRA and TOPSIS analysis showed that the comprehensive quality of S5 and S10 was relatively good. CONCLUSIONS The established method is accurate， stable and simple. Combined with multivariate statistical analysis method， it can be used for quality evaluation of P. forrestii. The quality of samples from Jiuzhou Town and Caiguan Town of Xixiu District in Anshun City of Guizhou Province are relatively good among 11 different origin samples.
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In this study, an ultra-performance liquid chromatography-quadrupole time-of-flight high resolution mass spectrometer(UPLC-Q-TOF-HRMS) was used to investigate the effects of the active ingredients in Periploca forrestii compound on spleen metabolism in rats with collagen-induced arthritis(CIA), and its potential anti-inflammatory mechanism was analyzed by network pharmacology. After the model of CIA was successfully established, the spleen tissues of rats were taken 28 days after administration. UPLC-Q-TOF-HRMS chromatograms were collected and analyzed by principal component analysis(PCA), orthogonal partial least squares discriminant analysis(OPLS-DA), and MetPA. The results showed that as compared with the blank control group, 22 biomarkers in the spleen tissues such as inosine, citicoline, hypoxanthine, and taurine in the model group increased, while 9 biomarkers such as CDP-ethanolamine and phosphorylcholine decreased. As compared with the model group, 21 biomarkers such as inosine, citicoline, CDP-ethanolamine, and phosphorylcholine were reregulated by the active ingredients in P. forrestii. Seventeen metabolic pathways were significantly enriched, including purine metabolism, taurine and hypotaurine metabolism, glycerophospholipid metabolism, and cysteine and methionine metabolism. Network pharmacology analysis found that purine metabolism, glycerophospholipid metabolism, and cysteine and methionine metabolism played important roles in the pathological process of rheumatoid arthritis. This study suggests that active ingredients in P. forrestii compound can delay the occurrence and development of inflammatory reaction by improving the spleen metabolic disorder of rats with CIA. The P. forrestii compound has multi-target and multi-pathway anti-inflammatory mechanism. This study is expected to provide a new explanation for the mechanism of active ingredients in P. forrestii compound against rheumatoid arthritis.
Subject(s)
Rats , Animals , Periploca , Cysteine , Cytidine Diphosphate Choline , Network Pharmacology , Phosphorylcholine , Metabolomics , Arthritis, Rheumatoid/drug therapy , Biomarkers , Glycerophospholipids , Methionine , Purines , Chromatography, High Pressure LiquidABSTRACT
Aim To investigate the therapeutie effect thritis (CIA) rats based on the variation of gut micro- of active ingredient group ( AIG) in the compounds biota.Methods CIA model rats were induced by in- from Periploca forrestii Schltr.on collagin-induced ar- jection of emulsion containing bovine type II collagen.anrl enzyme-linked immunosorbent assay ( ELISA) was used to investigate the changes of TNF-a and IL-6 in ankle joint of rats in each group.Meanwhile, the gut microbial communities were measured using 16s rRNA high-throughput sequencing technology and its function was predicted by KEGG.Results AIG could signifi¬cantly restrain inflammatory cell infiltration and tissue proliferation in synovial tissues of CIA rats,and allevi¬ate joint swelling while significantly reducing the con¬tents of IL-6 and TNF-a in ankle joint of CIA rats.The analysis of gut microbial communities showed that AIG could increase the relative abundances of Bacte- roidetes, Alloprevotella and Prevotella _9 , and decrease the relative abundances of 1 1 genera such as Dubosiel- la, Lachnospiraceae_NKAA 136_group, etc.in CIA rats.The functional prediction showed that AIG coulrl influ¬ence apoptosis and other signaling pathways.Conclu¬sions AIG can obviously alleviate joint swelling, re¬strain inflammatory cell infiltration and tissue prolifera¬tion in joint synovium of CIA rats,and repair the disor¬dered gut microbiota structure of CIA rats, while affecting its apoptosis pathway and restore the body immunity.
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ObjectiveTo analyze the effective components of Periploca forrestii against rheumatoid arthritis（RA）by targeting tumor necrosis factor （TNF）-α based on network pharmacology and experimental verification. MethodThe preliminary research of the research group found that the alcohol extracts of P. forrestii （CDLF and CQAF） had significant anti-RA activities，and 10 monomers with such activities were identified. The anti-RA activities of active monomers，CDLF， and CQAF were compared by the enzyme-linked immunosorbent assay （ELISA）with interleukin（IL）-6，nitric oxide （NO），IL-1β， and prostaglandin E2（PGE2）as indicators. Network pharmacology was employed to analyze the possible molecular mechanism of P. forrestii against RA. The targeting ability of P. forrestii chemical monomers to TNF-α was verified by TNF-α molecular docking，surface plasmon resonance （SPR）， and TNF-α-induced L929 injury model. ResultELISA showed that the anti-RA activities of CDLF and CQAF were significantly stronger than those of identified 10 active monomers. Network pharmacology analysis showed that the core targets of P. forrestii against RA were signal transducer and activator of transcription protein 3 （STAT3），TNF， and IL-6. Gene Ontology（GO） analysis revealed collagen catabolism，inflammatory response，positive regulation of nuclear factor kappa-B（NF-κB） transcription factor activity，and positive regulation of B cell proliferation. Kyoto Encyclopedia of Genes and Genomes （EKGG） pathway enrichment analysis demonstrated TNF signaling pathway，phosphoinositide 3-kinase（PI3K）/protein kinase B（Akt） signaling pathway，NF-κB signaling pathway，Toll-like receptor signaling pathway，mitogen-activated protein kinase（MAPK） signaling pathway， etc. Verification experiments by TNF-α molecular docking，SPR， and TNF-α-induced L929 injury model found that CDLF and CQAF had good binding activities and could manifestly antagonize TNF-α. However， the active components separated and identified from CDLF and CQAF did not show the same anti-TNF-α activity. ConclusionThe CDLF and CQAF of P. forrestii may treat RA by targeting TNF-α. The experiments found that the isolated chemical components had weaker binding activity to TNF-α than CDLF and CQAF. Meanwhile，the research group isolated chemical components with a minimum mass fraction of 0.25 ng·g-1 from P. forrestii， which suggested that the active components generated by binding to TNF-α with anti-RA activities were presumedly trace components .
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Objective:To study the correlation of eight chemical components in Miao medicine <italic>Periploca forrestii</italic> from different producing areas with the ecological and soil factors. Method:The contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， isochlorogenic acid B， isochlorogenic acid A， isochlorogenic acid C， procyanidin A<sub>2</sub>， and periplocin were simultaneously determined by ultra-performance liquid chromatography （UPLC）. The root soil samples from various producing areas were collected for testing various soil factors， followed by climate data extraction with ArcGIS and topographic data recording using GPS. SPSS 24.0 was employed to conduct the bivariate analysis and stepwise regression analysis of the eight chemical components in <italic>P. forrestii</italic> from different producing areas with the ecological and soil factors. Result:Stepwise regression equations of the content of eight chemical components against ecological and soil factors were established. The findings demonstrated that neochlorogenic acid was negatively correlated with precipitation in the coldest season and chlorogenic acid negatively correlated with precipitation in the driest month. Cryptochlorogenic acid was negatively correlated with precipitation in the coldest season and average temperature in the warmest season， but positively with selenium. Isochlorogenic acid B was mainly affected by soil factors. Specifically， it was positively correlated with available iron and molybdenum but negatively with total phosphorus and available phosphorus. Isochlorogenic acid A was positively correlated with molybdenum but negatively with the coefficient of variation of precipitation. Isochlorogenic acid C showed a positive correlation with exchangeable magnesium. Procyanidin A<sub>2</sub> exhibited a positive correlation with molybdenum and a negative correlation with available potassium. Periplocin was negatively correlated with the coefficient of variation of precipitation. Conclusion:The correlation between the eight chemical components of <italic>P. forrestii</italic> and the ecological and soil factors has been clarified， which will provide reference for the introduction， cultivation， and standardized planting of <italic>P. forrestii </italic>and also a theoretical basis for further research on its ecological and soil factors and quality formation mechanism.
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Objective: To establish a pharmacokinetic (PK)-pharmacodynamic (PD) model of Periploca forrestii. Methods: The right hindfoot footpad of rats were 0.1 mL complete Freund's Adjuvant (CFA) to establish adjuvant arthritis (AA) rat model. Rats were ig P. forrestii extract (87 g/kg, twice a day for 14 d) and blood were collected via tail vein at 5, 15, 30, 45 min, 1, 1.5, 2, 3, 4, 6, 8, 12, 24 h after the last administration. The concentrations of 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, 5-O-caffeoylquinic acid in plasma samples were detected by HPLC-mass spectrometry (HPLC-MS/MS) to obtain the drug concentration-time curve. The level of interleukin-1β (IL-1β), rheumatoid factor (RF), tumor necrosis factor-α (TNF-α) in plasma samples were determined by kit to obtain the time-effect curve. WinNonLin software was used to fit the PK parameters of P. forrestii, and the time-effect relationship was fitted to obtain the PD parameters. According to the PD parameters, the PK-PD model of P. forrestii was established. Results: The PK-PD model of P. forrestii according to the WinNonLin software was fitted in accordance with the Inhibitory Effect Sigmoid E0 model, in which the blood concentrations of 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, 5-O-caffeoylquinic acid could be calculated based on the potency values, and the potency values could be calculated based on the blood concentrations. Conclusion: There was a correlation between the concentrations of IL-1β, RF, TNF-α and 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid and 5-O-caffeoylquinic acid. These three components in P. forrestii extract could inhibit the secretion of IL-1β, RF and TNF-α to treat rheumatiod arthritis.
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Objective: To extract and purify total flavonoids from Periploca forrestii Schltr.(P. forrestii),and test the in vitro inhibitory activity of the total flavonoids and two flvaonoidal compounds in P. forrestii,so as to provide a reference for studies on the related medicinal substances in P. forrestii. Methods: Total flavonoids were extracted from P. forrestii and then purified by the column chromatography on macroporous resin and polyamide columns. The content of total flavonoids was determined according to the Lambert-Beer’s law. The in vitro xanthine oxidase(XOD)inhibitory ac- tivity was assayed for total flavonoids and the two flavonoidal compounds by the ultraviolet spectrophotometry. Results The purified total flavonoids had a content of more than 95%. The total flavonoids and two flavonoidal compounds all showed an inhibitory effect on XOD in vitro,with the inhibitory rate enhanced with increasing concentration. The IC50 of the total flavonoids as well as the two flavonoidal compounds,quercetin-3-O-α-L-pyranoside(QP)and quercetin-7-O-β- D-glucopyranoside(QG)were 608.9,221.2 and 261.2 μg/ml,respectively. Conclusion: The total flavonoids as well as the two flvaonoidal compounds QP and QG in P. forrestii all showed the in vitro inhibitory activity on XOD.
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Periploca forrestii, a traditional medicine commonly used by Miao people, is one of the "three treasures of Miao medicine", which mainly contains various components such as cardiac glycosides, flavonoids, ceramides, terpenoids, phenylpropanoid and volatile oils. It has significant pharmacological effects including cardiotonic, anti-inflammatory, antioxidant, pain-suppressing, and antibacterial activities, and is used to treat rheumatoid arthritis, bruises, stomach pain, dyspepsia, amenorrhea, and dysentery. Relevant domestic and abroad literatures were summarized, and a comprehensive review of the chemical constituents, pharmacological effects, clinical application, quality control and spectrum-effect relationship of Periploca forrestii was conducted, to provide evidences for further investigation of Periploca forrestii Schltr.
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Objective:To study the serum pharmacochemistry of Periploca forrestii rhizomes,and to investigate the pharmacological material basis of extract of P. forrestii rhizomes in rats. Method:Rapid identification of constituents absorbed into blood was carried out by UPLC-Q-TOF-MS,according to retention time,accurate relative molecular mass and standard substance comparison,these constituents were identified and speculated by Data Analysis,Metabolite Detect and other softwares,then preliminary determination of constituents absorbed into blood of rats after oral administration of extract of P. forrestii rhizomes was investigated. Result:Totally 17 constituents absorbed into blood were detected in serum,ten of them were prototype constituents and the other were metabolites.Seven of the prototypes were identified as 5-O-caffeoylquinic acid,4-O-caffeoylquinic acid,3-O-caffeoylquinic acid,3,4-di-O-caffeoylquinic acid,3,5-di-O-caffeoylquinic acid,4,5-di-O-caffeoylquinic acid and periplocin. Conclusion:These constituents absorbed into blood may be substances that act directly in vivo of P. forrestii rhizomes,and it is helpful to clarify pharmacological material basis and mechanism of this herb.
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OBJECTIVE:To establish the method for content determination of chlorogenic acid and periplocin in Miao medicine Periploca forrestii. METHODS:HPLC method was adopted. The determination was performed on Xtimate C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set at 220 nm,and column temperature was maintained at 25 ℃. The sample size was 10 μL. Cluster analysis was conducted according to the content of chlorogenic acid and periplocin in samples by SPSS 23.0 software. RESULTS:The linear range of chlorogenic acid and periplocin were 0.040 6-1.8 μg(r＝0.999 4)and 0.016 8-2.3 μg(r＝0.999 9),respectively. RSDs of precision,stability and repeatability tests were all lower than 5.0%. The quantitation limits were 0.918 0,0.084 3 μg/mL, and detection limits were 0.102 0,0.025 3 μg/mL, respectively. RSD of durability were lower than 3.0%. The recoveries were 102.66%-104.00%(RSD＝0.53%,n＝6),96.44%-100.79%(RSD＝1.73%,n＝6),respectively. RSD of durability were lower than 3.0%. The result indicated that 18 batches of samples were divided into 3 categories by cluster analysis. S2,S4,S10,S12 and S14-S18 were divided into one category;S1,S3,S5-S7,S9,S11 and S13 were divided into another category;S8 was regarded as one category. CONCLUSIONS:The method can be applied for quality control and evaluation of P. forrestii. The contents of chlorogenic acid and periplocin in P. forrestii from different producing areas are different greatly. There is a certain correlation between the content of each component and the producing area.
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Objective: To study the chemical constituents of Periploca Radix. Methods: The chemical constituents of Periploca forrestii were separated using various chromatographic techniques. Their structures were elucidated by spectral analysis. Results: Nineteen compounds were isolated from the 70% ethanol extract of P. forrestii, and identified as 3-O-caffeoylquinic acid methyl ester (1), 4-O-caffeoylquinic acid methyl ester (2), 5-O-caffeoylquinic acid methyl ester (3), 3-O-caffeoylquinic acid (4), 4-O-caffeoylquinic acid (5), 5-O-caffeoylquinic acid (6), 1, 3-di-O-caffeoylquinic acid (7), 3, 4-di-O-caffeoylquinic acid (8), 3, 5-di-O- caffeoylquinic acid (9), 4, 5-di-O-caffeoylquinic acid (10), protocatechuic aldehyde (11), p-hydroxybenzoic acid (12), o-hydroxybenzoic acid (13), syringic acid (14), vanillic acid (15), periforgenin A (16), Δ5-pregnene-3β, 17α, 20α-triol (17), periforgenin C (18), and periplogenin (19). Conclusion: Compounds 1-12 are isolated from the plants of genus Periploca Linn. for the first time, and compound 13 is isolated from P. forrestii for the first time.
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OBJECTIVE:To study the effects of caffeoylquinic acid derivative fractions (CADF) in Miao medicine Periploca forrestii on proliferation and secretion of inflammatory cytokines in tumor necrosis factor α(TNF-α)-induced human rheumatoid ar-thritis(RA)fibroblast-like synoviocytes MH7A,and explore its mechanism on anti-RA. METHODS:MH7A cells were divided in-to blank group,TNF-α model group,methotrexate group (positive control,20 mg/L) and CADF different mass concentrations groups(50,100,200,400 mg/L). Except for blank group,other groups received 50 μg/L of TNF-α to stimulate MH7A cells. Af-ter treated by suspension with TNF-α and related medicines for 24 h,the cell proliferation and contents of nitric oxide(NO),pros-taglandin E2 (PGE2),interleukin 1β(IL-1β),interleukin 6 (IL-6) in culture medium were detected. RESULTS:Compared with blank group,cell proliferation activity in TNF-αmodel group was significantly enhanced(P<0.01),contents of NO,PGE2,IL-1β, IL-6 in culture medium were significantly increased(P<0.01). Compared with TNF-α model group,cell proliferation in each ad-ministration group were significantly inhibited(P<0.05 or P<0.01),contents of NO,PGE2,IL-1β,IL-6 in culture medium weresignificantly decreased (P<0.01),showing certain dose-effect relationship with CADF. CONCLUSIONS:CADF can play the role in anti-RA by inhibiting the TNF-α-induced prolifera-tion of MH7A cells and reducing the secretion of inflammatory cytokines NO,PGE2,IL-1β,IL-6.
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OBJECTIVE:To study the effects of caffeoylquinic acid derivative fractions (CADF) in Miao medicine Periploca forrestii on proliferation and secretion of inflammatory cytokines in tumor necrosis factor α(TNF-α)-induced human rheumatoid ar-thritis(RA)fibroblast-like synoviocytes MH7A,and explore its mechanism on anti-RA. METHODS:MH7A cells were divided in-to blank group,TNF-α model group,methotrexate group (positive control,20 mg/L) and CADF different mass concentrations groups(50,100,200,400 mg/L). Except for blank group,other groups received 50 μg/L of TNF-α to stimulate MH7A cells. Af-ter treated by suspension with TNF-α and related medicines for 24 h,the cell proliferation and contents of nitric oxide(NO),pros-taglandin E2 (PGE2),interleukin 1β(IL-1β),interleukin 6 (IL-6) in culture medium were detected. RESULTS:Compared with blank group,cell proliferation activity in TNF-αmodel group was significantly enhanced(P<0.01),contents of NO,PGE2,IL-1β, IL-6 in culture medium were significantly increased(P<0.01). Compared with TNF-α model group,cell proliferation in each ad-ministration group were significantly inhibited(P<0.05 or P<0.01),contents of NO,PGE2,IL-1β,IL-6 in culture medium weresignificantly decreased (P<0.01),showing certain dose-effect relationship with CADF. CONCLUSIONS:CADF can play the role in anti-RA by inhibiting the TNF-α-induced prolifera-tion of MH7A cells and reducing the secretion of inflammatory cytokines NO,PGE2,IL-1β,IL-6.
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<p><b>OBJECTIVE</b>To determine the anti-inflflammatory effects of an ethanol fraction of Periploca forrestii Schltr. (EFPF) and to investigate the potential mechanisms underlying in vivo and in vitro models.</p><p><b>METHODS</b>The antiinflflammatory effects of EFPF were evaluated using the xylene-induced mouse ear edema and carrageenan-induced rat paw edema models in vivo. In vitro, RAW264.7 cells were exposed to 0-800 μg/mL EFPF and the cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Then cells were treated with different concentrations of EFPF (100-400 μg/mL) and stimulated with lipopolysaccharide (LPS, 1 μg/mL) for 24 h. The supernatant was analyzed for nitric oxide (NO) using the Griess reagent, and the levels of inflflammatory mediators and cytokines were determined using enzyme-linked immunosorbent assays for prostaglandin E(PGE), tumor necrosis factor α (TNF-α), interleukin (IL) 6, and IL-10. The protein expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor κB (NF-κB), and mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK were examined by Western blot.</p><p><b>RESULTS</b>Compared with the control group, EFPF signifificantly reduced mouse ear edema and rat paw edema rate (P<0.05 or P<0.01). Compared with the LPS group, EFPF signifificantly inhibited the LPS-stimulated production of NO, PGE, TNF-α and IL-6 (P<0.05 or P<0.01), and increased the IL-10 production (P<0.05). EFPF also signifificantly inhibited LPS-induced protein expressions of iNOS and COX-2, suppressed the phosphorylation and degradation of inhibitor of NF-κB-α, decreased p65 level, and inhibited the phosphorylation of p38, ERK1/2 and JNK P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>EFPF exerted anti-inflflammatory effect by reducing protein expressions of iNOS and COX-2 and the production of the inflflammation factors, including TNF-α, IL-6, NO and PGE, mainly through inhibition of LPS-mediated stimulation of NF-κB and MAPK signaling pathways.</p>
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Objective:To investigate the effects of periploca forrestii schltr in the treatment of acute gout arthritis.Methods:60 healthy male SD rats were equally randomly divided into 6 groups:normal control group( NC) ,model group( M group) ,colchicine group (C group),high doses group of periploca forrestii schltr(HD group),middle doses group of periploca forrestii schltr(MD group) and low doses group of periploca forrestii schltr( LD group).Except the normal control group,model of gouty arthritis was induced in other groups by uric acid salt,colchicine(positive control) and different dose of periploca forrestii schltr were given by intragastric ad minis-tration.Swelling dimension of joints were observed at 3,5,7 days after treatment.All rats were killed after 7 days of treatment and ankle joint tissue was taken for pathological examination and the peripheral blood of rats was prepared for detecting the expression of interleukin 1β(IL-1β),IL-6,IL-8 and tumor necrosis factor(TNF-α) using enzyme linked immunosorbent test(ELISA).Results:The ankle joint swelling of periploca forrestii schltr group was significantly lower than that in the model group,and the effect of high doses group was much better than the low doses group after 7 days treatment(P<0.05);compared with model group,the inflammatory cells of each treatment groups were decreased and high doses group did not differ from that of normal control group;the levels of IL-1β,IL-6,IL-8 and TNF-αin periploca forrestii schltr group were dramatically lower than those in the model group in a dose-dependent manner.Conclusion:Periploca forrestii schltr has good therapeutic effect in rats with acute gouty arthritis and shows a dose-dependent response,and the mechanism may relate to the inhibition of inflammatory cytokines expression.