Studies On Physicochemical Property And Bioactivities Of Turkish Gall Powder

Objective:By comparing the content of polysaccharides in the herb and residue of Turkish galls,combined with the previous studies on the determination of other components in the two,it was confirmed that there were still a large number of active ingredients in the residue.To enhance the dissolution and absorption of the active ingredients of Turkish galls of crude medicine resource,improved the curative effect and saved the resources of medicinal materials and expanded the ways of efficient utilization of Turkish galls,for providing scientific theoretical basis for further development and application of the Turkish galls ultrafine powder.1.Comparing the content of polysaccharides in the raw material of gall with that in the residue,and combining with the previous studies on the determination of other components in the residue,the optimum dissolution scheme of gall was worked out:powder of medicinal materials.2.Five kinds of Turkish galls powder(TGP)with different particle sizes were prepared and their powder characteristics were analyzed.3.Analysis of chemical composition and structure of five different particle sizes of TGP.4.the effects of five different particle sizes of TGP are compared by three antioxidant experimental models.5.To analyze the inhibitory effect of gall on three kinds of colon cancer cells;6.To explore the autophagic effect of gall on NRK-GFP-LC3 cells.Methods:1.The content of polysaccharides in gall was determined by ultraviolet-visible spectrophotometry(UV-Vis).2.Five kinds of gall powder with different particle sizes TGP were prepared by vibratory cell-level ultrafine grinding technology(TY-L8 vibratory cell-level ultrafine grinder).The particle size distribution was measured by micrometer laser particle size analyzer.3.The ultrafine gall powder with different particle sizes was detected by powder characteristics tester.The repose angle,sliding angle,collapse angle,bulk density and compacting density of Micro-powders are the characteristics of powders.4.The dissolution of TGP with five different particle sizes was determined at 40,55,70,85 and 100?C respectively.5.Microscopic analysis:observed the characteristics of TGP under fluorescence inversion microscope;6.SEM analysis:observed the shape differences of five different size TGP,as well as the morphological differences between traditional and ultrafine grinding particles;7.The content differences of gallic acid,methyl gallate and ellagic acid in five different particle sizes of TGP extracts were determined and compared by reverse high performance liquid chromatography(RP-HPLC).8.IR analysis:the structural differences of five different particle sizes of TGP and its corresponding extracts were compared;9.DPPH,hydroxyl radical and superoxide anion were established.The antioxidant activity of powders with different particle sizes was compared by radical scavenging experiment and the correlation between the antioxidant activity and the related chemical components was analyzed.10.MTT colorimetric method was used to study the inhibitory effect of three different types of colon cancer cells(HCT-116,SW480,Caco-2).11.Laser scanning confocal microscopy(LSCM)was used to observe the three-dimensional morphological changes of NRK-GFP-LC3 cells after administration and to analyze the phenomenon of autophagic apoptosis.Results:1.The extraction rate of Polysaccharides from 10 batches of residue of Turkish galls was 1.05-1.24%,and the extraction rate of Polysaccharides from 10 batches of Turkish galls was 4.49-5.53%.Combining with the previous research results on other components of the two,it also shows that there are more residual components in the residue.The particle sizes of TGP were treated by a vibrating-type ultrafine grinding treatment with particle sizes of>450,400�250,250�100,100�50,and<50?m.The physicochemical properties and powder properties of TGP were evaluated.2.According to the experimental results of physical properties of powders,the bulk density(0.68-0.32g/cm~3)and tap density(0.83-0.70 g/cm~3)decrease with the decrease of particle size,while the angles of repose(35.43-56.28 degrees),slid(35.54-59.23 degrees),crash(35.68-56.28degrees)increase with the decrease of particle size.Total ashs content decreased from2.34%to 1.75%,but moisture and water solubility index increased.3.The contents of gallic acid(9.47 mg/g),methyl gallate(34.78 mg/g)and ellagic acid(0.79 mg/g)were the highest in the TGP extracts with diameter less than 50 um.4.The antioxidant activity of DPPH,hydroxyl radical and superoxide anion radical scavenging experiment was the best when the particle size was less than 50 um.(p<0.05).5.In addition,FTIR analysis shows that with the decrease of powder size,the structure of the main components in the powder is not destroyed by the treatment of vibrating-type ultrafine grinding.6.The SEM images clearly show the different particle size states,and the particle morphologies of traditional grinding and vibrating-type ultrafine grinding.Microscopic observations revealed powder properties of catheters,oil cells,starch granules and crystal fibers.7.MTT assay showed that gall extract and GA,MG and EA could inhibit the proliferation of colon cancer cells in the corresponding concentration range.Under the 48 hours of administration,the results were statistically significant.8.LSCM was used to observe the effect of GA,MG and EA on the expression of autophagy in NRK-GFP-LC3 cells.From the perspectives of transverse drug concentration change,longitudinal drug duration change and overall drug type difference,it was found that these three factors had a common effect on the autophagy of NRK-GFP-LC3 cells.Conclusion:In order to further improve the utilization rate of Turkish galls herb resources,ultrafine grinding technology was used to optimize the extraction particle size scheme of Turkish galls,and the physical and chemical properties and biological activities of TGP were evaluated.Research.It was also proved that Turkish galls extract could inhibit the growth of colon cancer cells and achieve anti-cancer effect by inducing autophagy and apoptosis of colon cancer cells.