| Cyathulae radix is the dry root of Cyathula officinalis Kuan.Its name first appeared in the Tang Dynasty,Lin daoren Xian chou li shang xu duan mi fang,Cyathula officinalis distributed in Sichuan,cloud,Guizhou and other provinces,to primary production in Sichuan and its name,its sweet,slightly bitter,flat,liver,kidney,with liver and kidney,strong bones and muscles,eliminate swelling and other effects.In the early stage of the research group,the constituents belonging to Cyathula officinalis Kuan were assigned to taste,and the polysaccharide component and phytosterone component were determined to be sweet and warm,and the 20%alcohol wash component and the phytosterone component were bitter and cool,but had repair liver and kidney of sweetness strong bones and muscles of bitterness for mechanism of pharmacodynamics have not yet been development,which limits the interpreta tion of the basic scientific connotation of the theory of the taste of Cyathula officinalis Kuan.Therefore,in this study,sweet polysaccharide components and bitter components were selected for in-depth study to provide theoretical basis for the correct guidance of clinical practice and scientific research.This article carried out the following researches on the sweet polysaccharides and bitter phytosterone of Cyathula officinalis Kuan.1.Literature review.To taste as the center,through a large number of domestic and foreign literature retrieval system to clarify the traditional TCM and taste of Cyathulae Radix name,variety,development of origin and the process of evolution and of Cyathula root chemical composition,current situation in the research of pharmacological action and clinical application,understand near modern flavor research theory and Cyathulae Radix pharmacology and chemical composition research results,summarize the clinical application,complete the review of research on the theory of traditional TCM taste and Cyathulae Radix for the experimental research of the development lay the foundation for the literature of rich and effective.2.Study on the separation of sweet polysaccharides from Cyathula officinalis Kuan.The total polysaccharides of Cyathula officinalis Kuan were extracted by water decoction and alcohol precipitation,and then separated and purified by weakly weak cation exchange resin and ion exchange cellulose(Cellulose DE-52).HPLC detect molecular of polysaccharide from Cyathulae Radix split components in separation and purification of 6 kinds of homogeneous polysaccharides:CP-1-CP-6.The sugar composition analysis was used to determine the monosaccharide composition of the six homogeneous polysaccharides.In addition,the spatial morphologies of two homogeneous polysaccharides were studied by atomic force microscopy(AMF).Research indicates:The molecular weights of RCPS1,RCPS2,RCPS3,RCPS4,RCPS5,and RCPS6 were 6.01 × 106 Da,8.64×107 Da,3.40×107 Da,4.15×107 Da,>7.56×107 Da,and>9.74×107 Da,respectively.The constituent monosaccharides of RCPS1 are mannose,galactose,and glucose in a molar percentage of 0.71:32.6:2.40.The composition monosaccharide of RCPS 2 is mannose,galactose,and glucose,and its molar percentage is 0.28:19.9:0.16,RCPS3.The composition of the monosaccharide is glucuronic acid,glucose,its molar percentage is 3.34:0.14,the composition of RCPS4 monosaccharide is mannose,galacturonic acid,glucose,its molar percentage is 0.13:0.97:0.09,the composition of RCPS5 single The sugar is galacturonic acid,glucose,its molar percentage is 1.82:1.21,and the constitutional monosaccharide of RCPS6 is rhamnose,galacturonic acid,glucose,arabinose,and its molar percentage is 0.35:2.11:0.85:0.18.Atomic force microscopy studies the spatial morphology of RCPS1 and RCPS5.The RCPS1 and RCPS5 presented spherical structures with diameters of 34.36 nm and 33.33 nm.The lumps of irregularly shaped lumps in the three-dimensional view of the AFM resembled tens of thousands of helminths,suggesting that the RCPS1 and RCPS5 molecules can undergo free-formed coiling.It may be related to its unique branching structure.3.The pharmacological study of sweet polysaccharides based on Cyathula officinalis KuanCyathula officinalis Kuan has the effect of supplementing liver and kidney,and it establishes a model of acute liver injury in rats by CCl4 induction method,which proves that the sweet polysaccharide-based polysaccharides of Cyathula officinalis Kuan has anti-acute liver injury.Serum inflammatory cytokines IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay(ELISA).The changes of serum ALT and AST activity were used to demonstrate whether the model was established successfully.The activities of oxidative factors MDA,SOD and GSH-PX were measured.Rat thymus index,spleen index.Liver histopathological changes were observed by H&E staining.Using immunohistochemical staining and Western blot to study its role in the pathway.Achyranthes bidentata polysaccharide has anti-acute liver damage effect.The performance of Achyranthes bidentata inhibited the levels of serum ALT and AST,decreased the content of MDA,restored the activity of SOD and GSH-PX,and reduced the production of inflammatory cytokines TNF-α and IL-6.In Cyathula officinalis Kuan polysaccharides group,the thymus index and spleen index were significantly increase,reflecting the anti-inflammatory and immune-enhancing effects of sweet polysaccharides of Cyathula officinalis Kuan.Liver histopathological changes were observed by H&E staining.Using immunohistochemical staining and Western blot to study its role in the pathway.Cyathula officinalis Kuan polysaccharide has anti-acute liver damage effect.The performance of Cyathula officinalis Kuan inhibited the levels of serum ALT and AST,decreased the content of MDA,restored the activity of SOD and GSH-PX,and reduced the production of TNF-α and IL-6.In the H&E liver tissue staining,Cyathula officinalis Kuan polysaccharides were found to reduce the extent and extent of liver lesions and reduce the infiltration of inflammatory cells.The results of immunohistochemistry showed that Cyathula officinalis Kuan polysaccharide inhibited the expression of TLR4 and NF-κBp65,Western blot results showed that inhibited the activity and migration of phosphorylated iκBα,iκBα,NF-κBp65 and TLR4,Explain the possible role of Cyathula officinalis Kuan polysaccharide against acute liver injury.The first time further study anti-acute liver injury effect of Cyathula officinalis Kuan polysaccharides,which proves the natural basis of sweet taste of Cyathula officinalis Kuan is the correctness and objectivity of polysaccharides,and it can provide a theoretical basis for replacing or partially replacing traditional anti-acute liver injury drugs in the future.in accordance with.4.The chemical study on the phytosteronesof the bitter taste of Cyathula officinalis KuanThe phytosterone components of Cyathula officinalis Kuan were extracted by methanol reflux method.The phytosterone components of Cyathula officinalis Kuan were separated and purified by SP825L macroporous adsorption resin,ODS column chromatography,preparative liquid chromatography and recrystallization.1H-NMR and 13C-NMR mass spectrometry tests were used to determine the structure of the compound.Research result:We isolated 12 phytosterone compounds.The structures of 12 phytostero ne compounds were identified by 1H-NMR,13C-NMR,mass spectrometry and other spectroscopic tests.24-hydroxycyasterone(1),25-Epi-28-epi-cyasterone(2),24,28-diepi-cyasterone(3),caliponone(4),28-epi-cyasterone(5),neosterone B(6),neosterone A(7),Makisterone B(8),Makisterone A(9),Capitasterone(10),Precyasterone(11),Isocyasterone(12).5.Study on the Pharmacokinetics of bitter phytosterone of Cyathula officinalis KuanCyathula officinalis Kuan enters the blood stream,and it is good for the downside.It has the effect of strong bones and bones.The rat model of acute gouty arthritis was established by MSU induction,demonstrating that the phytosterone of Cyathula officinalis Kuan has anti-acute gouty arthritis effect.Observe joint swelling in rats.The concentration of uric acid,NO,IL-1β,TNF-α and the concentration of XOD in serum were measured by enzyme-linked immunosorbent assay(ELISA).Cyathula officinalis Kuan has anti-acute gouty arthritis effect.Cyathula officinalis Kuan phytosterone has anti-acute gouty arthritis effect.In Cyathula officinalis Kuan phytosterone group,the uric acid concentration was decreased in serum,which shows that the bitter taste of Cyathula officinalis Kuan has diuretic effect;It also inhibited the production of XOD,NO,IL-1β,TNF-α;Synovial tissue was observed by light microscopy that Cyathula officinalis Kuan can significantly reduce inflammatory cell infiltration,synovial cell proliferation,capillary hyperemia,and edema in the synovial tissue;The results of immunohistochemistry showed that Cyathula officinalis Kuan phytosterone inhibited the expression of NF-κBp65 and NALP3.Western blotting results showed that the expression of TLR4,NF-κBp65,COX-2,and NALP3 proteins.Explains the possible pathway of anti-acute gouty arthritis in Cyathula officinalis Kuan phytosterone.This article for the first time through the anti-acute arthritis of Cyathula officinalis Kuan,further proves that the bitter substance basis of Cyathula officinalis Kuan is correct and objective.Chuankuinone may be expected to become an anti-acute gouty arthritis drug.For the future to develop a Chinese medicine with good curative effect,little side effects and possibly multiple target effects,it provides the basis for scientific basis and clinical application value.The innovativeness and features of this study:In this study,chemical and pharmacological studies on basic sweet substances of polysaccharides and bitter substances of Cyathula officinalis Kuan phytosterone.It was demonstrated for the first time that the sweet polysaccharides of Cyathula officinalis Kuan had anti-acute liver damage,and it was further confirmed by chemical analysis and pharmacological evaluation that the task group was correct and objective of polysaccharide belonging to the sweet taste basis of Cyathula officinalis Kuan.It was demonstrated for the first time that the Cyathula officinalis Kuan phytosterone has anti-acute gouty arthritis effect,and further confirmed by chemical analysisand pharmacologic al evaluation that the basis for the bitter taste of the Cyathula officinalis Kuan phytosterone is the correctness and objective.It also show more confirmed the"traditional TCM also has(gas)and taste.(gas)and the taste is Chinese traditional medicine simultaneously two kinds of characteristics of the specific efficacy of herbs and drugs.Drug(gas)is drugs through different pathways to mainly affect the body’s energy metabolism,metabolism characteristics and traditional TCM taste with split,combination,traditional TCM blindly of,the objectivity of I-Dozer x y(y=x)of traditional TCM taste theory hypothesis.And further improve the drug belonging to the judgment method,has the important theory and application value. |
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