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In this study, the crude polysaccharides was extracted from Shengfupian and purified by Sevag deproteinization. Then, the purified neutral polysaccharide fragment was obtained by the DEAE-52 cellulose chromatography column and Sephadex G-100 co-lumn. The structure of polysaccharides was characterized by ultraviolet spectroscopy, infrared spectroscopy, ion chromatography, and gel permeation chromatography. To investigate the anti-inflammatory activity of Shengfupian polysaccharides, LPS was used to induce inflammation in RAW264.7 cells. The expression of the CD86 antibody on surface of M1 cells, the function of macrophages, and the content of NO and IL-6 in the supernatant were examined. An immunodepression model of H22 tumor-bearing mice was established, and the immunomodulatory activity of Shengfupian polysaccharides was evaluated based on the tumor inhibition rate, immune organ index and function, and serum cytokine levels. Research indicated that Shengfupian polysaccharides(80 251 Da) was composed of arabinose, galactose, glucose, and fructose with molar ratio of 0.004∶0.018∶0.913∶0.065. It was smooth and lumpy under the scanning electron microscope. In the concentration range of 25-200 μg·mL~(-1), Shengfupian polysaccharides exhibited little or no toxicity to RAW264.7 cells and could inhibit the polarization of cells to the M1 type and reduce the content of NO and IL-6 in the cell supernatant. It could suppress the phagocytosis of cells at the concentration of 25 μg·mL~(-1), while enhancing the phagocytosis of RAW264.7 cells within the concentration range of 100-200 μg·mL~(-1). The 200 mg·kg~(-1) Shengfupian polysaccharides could alleviate the spleen injury caused by cyclophosphamide, increase the levels of IL-1β and IL-6, and decrease the level of TNF-α in the serum of mice. In conclusion, Shengfupian polysaccharides has anti-inflammatory effect and weak immunomodulatory effect, which may the material basis of Aconm Lateralis Radix Praeparaia for dispelling cold and relieving pain.
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Animales , Ratones , Interleucina-6/genética , Citocinas/metabolismo , Polisacáridos/química , Antiinflamatorios/química , Espectrofotometría InfrarrojaRESUMEN
Objective:To explore the material basis and mechanism of Sangjiang Ganmao injection （SG） in the treatment of common cold by ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） and network pharmacology. Method:UPLC-Q-Orbitrap HRMS was used to identify the chemical components of SG with mobile phase of acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-10 min， 4%-15%A； 10-35 min， 15%-30%A； 35-45 min， 30%-33%A； 45-55 min， 33%-60%A； 55-58 min， 60%A）， flow rate of 0.2 mL·min<sup>-1</sup>， electrospray ionization （ESI） and scanning range of <italic>m</italic>/<italic>z</italic> 100-1 500 under positive and negative ion modes. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and GeneCards 5.0 database were used to screen and predict the potential targets of chemical components in SG， STRING 11.0 database and Cytoscape 3.7.2 software were used to construct protein-protein interaction （PPI） network model， gene ontology （GO） analysis and pathway analysis were performed on potential targets by Metascape 3.5， Reactome database and Kyoto Encyclopedia of Genes and Genomes （KEGG）， Cytoscape 3.7.2 software was used to build the network of "herbs-ingredients-key targets". Result:A total of 54 components in SG were identified， and 80 potential targets of SG for treatment of common cold were predicted and screened based on this. SG exerted therapeutic effects by acting on targets such as interleukin （IL）-6， tumor necrosis factor （TNF） and IL-10， and signaling pathways such as IL-17 signaling pathway， TNF signaling pathway and interaction of cytokine receptors. Conclusion:SG may interfere with the expression of inflammatory cytokines by acting on related targets and pathways such as inflammation and immune system， and regulate the immune function of the body as a whole， thereby exerting a therapeutic effect.
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Objective:To establish and apply a new practical analytical method for identifying the fishy odor of Cordyceps based on headspace-solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry （HS-SPME/GC-QQQ-MS/MS） technique. Method:The InertCap Pure-WAX capillary column （0.25 mm×30 m， 0.25 μm） was used for chromatographic separation. The injection port temperature was set at 250 ℃. The injection mode was split injection with a ratio of 5∶1. High purity helium was used as the carrier gas and control mode was set to constant pressure. The column flow rate was 1.43 mL∙min<sup>-1</sup>， the linear velocity was 43.3 cm∙s<sup>-1</sup>， and the purge flow rate was 3.0 mL∙min<sup>-1</sup>. The chromatographic column temperature program as follows：maintained the initial temperature at 50 ℃ for 5 min， and increased the temperature at a rate of 10 ℃∙min<sup>-1</sup> to 250 ℃， held for 10 min. The column equilibrium time was 2.0 min. The ion source of mass spectrographic analysis was electron ionization with ion source temperature of 200 ℃， and the monitoring mode was set to multiple reaction monitoring. Result:Seven batches of Cordyceps samples were collected， including 3 batches from Sichuan， 3 batches from Qinghai and 1 batch from Tibet. There were six batches of counterfeits， including 3 batches from Sichuan， 2 batches from Guizhou and 1 batch in Xinjiang. A total of 81 volatile compounds were screened out in Cordyceps， which could be divided into 13 types （esters， ketones， aldehydes and others） according to the compound structure， indicating that the fishy odor of Cordyceps was a complex odor. There was no significant difference in the types of volatile compounds of Cordyceps from different regions， which suggested that these volatile compounds in Cordyceps produced in Tibet （Naqu）， Qinghai （Yushu and Guoluo） and Sichuan （Litang， Rangtang and Seda） were relatively consistent. However， the contents of some volatile compounds in Cordyceps produced in different regions were quite different， and 16 volatile compounds with significant difference were screened out， including 1-methoxy-2-propyl acetate， <italic>γ</italic>-octalactone， hexyl acetate and others， those compounds maybe could been used as the quality markers for identification of regions of Cordyceps. There was a large difference in volatile compounds between Cordyceps and its counterfeits， and 34 volatile compounds were screened out， including ethyl acetate， acetophenone， 2-ethyl-1-hexanol and others， those compounds maybe could been used as the quality markers for authenticity identification of Cordyceps. Conclusion:In summary， the established method for identifying the fishy odor of Cordyceps in this paper has the characteristics of high sensitivity， accuracy and simplicity， which can provide reference for the analysis of volatile compounds in other Chinese herbal medicines.
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Periplaneta americana is one of the important basic medicinal materials of traditional Chinese medicine "fei lian". The traditional functions mainly include promoting blood circulation， sore muscles， diuresis， spleen and phlegm. Because of its exact curative effect， proprietary Chinese medicines， which are mainly used as raw materials， are widely used in clinical practice， especially in the repair of various wounds. The drug has not been included in the Chinese Pharmacopoeia. The local standard is only based on the alanine content of total amino acids. The physiologically active small peptides， nucleosides， proteins and other substances have not been obtained. Qualitative or quantitative control. In recent years， peptide monomers isolated from the P.americana， such as antimicrobial peptides， neuropeptides， and diuretic peptides， have strong pharmacological activities such as antibacterial， antitumor， and muscular neurotrophic， and dihydroisocoumarins are also irritating. Dermal Dermal fibroblasts produce collagen. Based on this， this paper uses CNKI， Wanfang Database and Pubmed Database to search the relevant research literatures of P.americana from 1984 to 2019， and systematically analyzes the current research of P.americana from three aspects： chemical composition-pharmacological action-clinical application. Interpretation provides reference for the further development of the drug and the development of more specific and stable quality control standards for its proprietary Chinese medicines.
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Objective:To prepare Periplaneta americana thermosensitive hydrogel and investigate its effect on wound healing in diabetic rats. Method:Taking N-isopropylacrylamide (NIPAM) and acrylic acid (AAc) as monomers, thermosensitive poly(NIPAM-co-AAc) [P(NIPAM-co-AAc)] polymeric material was prepared by free radical polymerization, then thermoresponsive copolymer P(NIPAM-co-AAc)-g-HA was synthesized by conjugating P(NIPAM-co-AAc) to hyaluronic acid (HA). The structure and lower critical solution temperature (LCST) of the graft copolymer were characterized by proton nuclear magnetic resonance spectroscopy (1H-NMR) and ultraviolet spectrophotometry (UV). P. americana thermosensitive hydrogel was prepared by dialysis method, and it was characterized by scanning electron microscope (SEM), rotation rheometer and thermogravimetric analyzer to observe section structure, rheological properties and thermal stability. Differential scanning calorimetry, X-ray diffraction and Fourier transform infrared spectroscopy were employed to identify the inclusion of P(NIPAM-co-AAc)-g-HA temperature sensitive material for P. americana extract, and to investigate the effect of P. americana thermosensitive hydrogel on wound healing in diabetic rats, and the rate of wound healing was calculated by Image-Pro Plus 6.0 software. Hematoxylin-eosin (HE) and Masson staining were used to observe the pathological changes of the wounds of rats in each group. Result:P(NIPAM-co-AAc)-g-HA temperature sensitive material was successfully synthesized, its LCST was between 29 ℃ and 31 ℃, it had a dense and uniform porous structure and could uniformly include P. americana extract. Pharmacodynamic studies showed that P. americana thermosensitive hydrogel group had the best effect on promoting wound healing, its infiltration degree of inflammatory cells was significantly reduced, collagen and fibroblasts arranged neatly and compactly, and the density of neovascularization was significantly increased by comparing with the model group. Conclusion:P. americana thermosensitive hydrogel can effectively promote wound healing of diabetic rats and overcome the shortage of marketed P. americana liquid preparations, this paper can provide a reference for the development of P. americana extract preparations to promote wound healing in diabetic patients.
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According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.
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Animales , Femenino , Humanos , Masculino , Ratones , Aconitum , Química , Alcaloides , Química , Toxicidad , Antiinflamatorios , Química , Toxicidad , Artritis Reumatoide , Quimioterapia , Cromatografía Líquida de Alta Presión , Evaluación Preclínica de Medicamentos , Medicamentos Herbarios Chinos , Química , Toxicidad , Hojas de la Planta , Química , Raíces de Plantas , Química , Tallos de la Planta , Química , Espectrometría de Masas en TándemRESUMEN
According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.
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Animales , Femenino , Humanos , Masculino , Ratones , Aconitum , Química , Alcaloides , Química , Toxicidad , Antiinflamatorios , Química , Toxicidad , Artritis Reumatoide , Quimioterapia , Cromatografía Líquida de Alta Presión , Evaluación Preclínica de Medicamentos , Medicamentos Herbarios Chinos , Química , Toxicidad , Hojas de la Planta , Química , Raíces de Plantas , Química , Tallos de la Planta , Química , Espectrometría de Masas en TándemRESUMEN
Aim To study the effect of the Kangfuxin liquid on 2,4,6-trinitrobenzene sulfonic acid (TNBS) induced ulcerative colitis in rats, and to explore its mechanism. Methods Male SD rats were divided into normal control, model control, SASP groups, and Kangfuxin low,medium and high dose groups. In ad-dition to the normal control group, other groups were induced ulcerative colitis with TNBS solution. Disease activity index (DAI), organ index, colon mucosa damage index (CMDI) and histopathological score (HS),the expression levels of IL-4, IL-17 in serum, and MPO, EGF, TGF-β1 in colonic mucosa were de-termined. Results The DAI score showed that the model was successful. Compared with the normal group,the level of IL-4 and IL-17, EGF and TGF-β1 in the model group were reduced significantly, while the CMDI score,HS score,colon index and MPO were elevated significantly. The DAI, CMDI, HS and MPO were reduced (P<0.05 or P<0.01), and the levels of IL-4, IL-17, EGF and TGF-β1 increased signifi-cantly(P <0.01). Conclusions Kangfuxin liquid can effectively alleviate ulcerative colitis induced by TNBS in rats. The mechanism may be related to the down-regulation of MPO expression and up-regulation of IL-4,IL-17,EGF and TGF-β1 levels.