| Codonopsis pilosula,as a kind of Chinese herbal medicine,is an authentic medicinal material in Gansu.It has important pharmacological effects such as anti-oxidation,regulation of the digestive system,anti-tumor,and enhancement of the body’s immunity,among which polysaccharides are very important active ingredient.In this paper,the crude polysaccharides of C.pilosula were extracted by hot water extraction method,and the protein was removed by Sevage method.After deproteinization,the extract was purified by DEAE-52 cellulose column chromatography and Sephadex G-100 gel column chromatography.Six different fractions of C.pilosula polysaccharides wcp1,wcp2,wcp3,wcp4,wcp5 and wcp6 were obtained by periodic acid oxidation.Fourier infrared spectroscopy(FTIR),Congo red,differential scanning.The molecular structure of Codonopsis pilosula polysaccharide was determined and analyzed by thermogravimetric analysis(TG),X-ray diffraction(XRD),circular dichroism(CD)and other methods.The anti-oxidation ability of Codonopsis pilosula polysaccharide was analyzed through the experiments on the scavenging and reducing ability of ABTS+· radical,DPPH· radical and hydroxyl radical..Finally,conventional methods were used to prepare C.pilosula polysaccharide into microcapsules.XRD,TG,FT-IR,atomic force microscopy(AFM)and scanning electron microscope(SEM)were used to characterize the microstructure of the microspheres.The drug loading and we investigated the effect of C.pilosula polysaccharide microcapsules on wound healing and scar repair by establishing a rat wound model.The results of the study are as follows:The relative molecular masses of wcp1,wcp2,wcp3,wcp4,wcp5,and wcp6 were 547.00,386.30,916.40,578.30,305.80 and 992.40 KDa.Among them,the molecular weight distribution of wcp5 is the widest and that of wcp4 is the smallest.Through the analysis of the monosaccharide composition of the polysaccharides from different parts of C.,the main components are mannose,glucose,and arabinose.Among them,wcp6 is mainly composed of mannose and galactose.Both wp4 and wcp5 contain small amounts of rhamnose and xylose.wcp2 contains a small amount of glucuronic acid.Infrared spectroscopy showed that the six different fractions of C.pilosula polysaccharides are mainly in β configuration.Through periodic acid oxidation experiments,it can be inferred that wcp1 and wcp3 may contain 1→2,1→3,and 1→2.6 types of glycosidic bonds.This experiment conducted a preliminary study on the sugar chain conformation of different fractions of C.pilosula polysaccharides through the Congo red experiment and CD,it can be seen that they all have a triple helix structure.SEM results show that the surface of C.pilosula polysaccharides wcp1,wcp2,and wcp4 showed smooth fiber flocs,while wcp3 and wcp6 showed uneven flocculent with small particles and filaments connected.wcp5 has a more obvious honeycomb structure.The research on the antioxidant activity of C.pilosula polysaccharide,that the scavenging effects of the polysaccharides DPPH· free radicals,ABTS+· free radicals,hydroxyl free radicals and other free radicals were measured.It is found that among the six different grades at a concentration of 0-10.0 mg/m L,the best scavenging effect on DPPH· free radicals is wcp6,and the best scavenging effect on ABTS+· radicals is wcp6 as well,accompanied by six different levels.Among the fractions,the best scavenging effect on hydroxyl radicals is wcp5.The scavenging ability of various free radicals is in a dose-dependent relationship,and the scavenging ability of free radicals of each level of polysaccharides is weaker than that of Vc at the same concentration.In addition,the reducing power of the six fractions of polysaccharides was measured,and it was found that under the same concentration conditions,wcp4 had the strongest reducing power and wcp1 had the weakest reducing power.The structure and physicochemical characteristics of C.pilosula polysaccharide microcapsules were studied,and the particle size distribution of C.pilosula polysaccharide microcapsules was relatively uniform.About 75.58% of its particle size is distributed in the range of 50-120 nm,and the average particle size is 92.92 nm.The prepared C.pilosula polysaccharide microcapsule has a drug loading rate of42.8%,an encapsulation rate of 59.92%,and a swelling rate of up to 250%.Routine blood test(external application for 14 days)results showed that C.pilosula polysaccharide microcapsules can promote the formation of new blood vessels in the trauma model,increase the content of hydroxyproline in the wound site,reduce the level of lipid peroxidase,and have strong antioxidant activity,indicating that the polysaccharide microcapsules of C.pilosula can promote wounds healing has a positive effect.By detecting the transcription levels of VEGF mRNA and miRNA genes,it was found that the C.pilosula polysaccharide microcapsule group could significantly increase the transcription levels of VEGF mRNA and miRNA,indicating that it plays a certain effective role in vascular remodeling during wound healing.Experiments show that C.pilosula polysaccharide microcapsules can exert great potential in wound healing treatment.In this paper,the research results of the physicochemical properties,antioxidant activity,and the preparation of microcapsules of C.pilosula polysaccharide to promote wound healing provide an important basis for the further development and utilization of C.pilosula polysaccharide and the comprehensive utilization of resources of C.pilosa. |
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