| ObjectTo find out the optimal matching range of active ingredients in Fructus aurantii,enrich the research content of formation factors of genuine medicinal materials,explore the distribution of genuine medicinal materials in Fructus aurantii,and elaborate the"genuine"of Fructus aurantii,so as to provide scientific basis for the safe and effective use of traditional Chinese Medicine.Method1.Forecast the suitable growth area of Fructus aurantii:the geographical distribution points of Fructus aurantii were obtained from the actual samples,digital herbarium data and literature data.The coordinate points were screened by Max Ent model and Arc GIS software,and the climate,soil and other environmental factors affecting the growth of Fructus aurantii were collected to obtain the most suitable ecological factors,determine the ecological environment for the growth of Fructus aurantii,and calculate the potential spatial distribution of Fructus aurantii.The ecological suitability was used to study the regionalization of Fructus aurantii.2.Animal experiment screening active components of Fructus aurantii:the gastrointestinal motility test of normal mice was used to screen the flavonoids and coumarins in Fructus aurantii.Extraction and separation of Polysaccharides from Fructus aurantii.The primary structure of Fructus aurantii polysaccharide was analyzed by different chromatographic methods.The structure-activity relationship between Fructus aurantii polysaccharide and its pharmacological action was established by normal and model animal experiments.3.Analyze the differences of chemical components in Fructus aurantii from different areas:establish the determination method of flavonoids and coumarins in Fructus aurantii,and the structure analysis method of polysaccharides.Based on environmental factors,analyze the differences of flavonoids and coumarins in Fructus aurantii from different areas and the structure differences of polysaccharides in Fructus aurantia.4.Selection of the best compatibility ratio:the method of multivariate statistical analysis was used to classify the Fructus aurantii,to obtain the sample information of each group,to fit the compatibility ratio of the active ingredients in the representative herbs of each group,and to infer the optimal compatibility ratio of the active ingredients combined with the pharmacological experiment verification.5.Pharmacodynamic experiment verification of the best matching range:establish the experimental model of spleen deficiency rats,the best matching ratio of active ingredients,fit the effect of the composition and its original medicinal materials on the model rats,analyze the key indicators of gastrointestinal motility,verify the best matching range of active ingredients,and improve the evaluation standard of genuine medicinal materials.6.To describe the quality of Fructus aurantii:Based on the optimal range of active ingredients,cluster analysis of the active components of Fructus aurantii from different places,find out the samples similar to the optimal matching results,and then explain the distribution of the real estate area and the traditional Chinese medicine of Fructus aurantii.Result1.The model established in this study is reliable and accurate,the results show that the main environmental factors affecting the growth and distribution of Fructus aurantii are temperature factor(solar horizontal radiation,annual average temperature,isothermal temperature,the lowest temperature in the coldest month,the average temperature in the driest season,the highest temperature in the warmest month),water factor(precipitation in the warmest season and wet season),and soil factor(soil clay content),some regional ecological factors in Jiangxi and Hunan show high similarity and stability.The potential distribution area of Fructus aurantii is estimated to be 211071 square kilometers,and the high suitable areas are Jiangxi(5.0×10~4 square kilometers),Hunan(5.9×10~4 square kilometers),Sichuan(4.3×10~4 square kilometers),Chongqing(3.1×10~4 square kilometers)and Zhejiang(0.8×10~4 square kilometers)And so on.The main production area of Fructus aurantii is basically consistent with the high fitness area of the original plant of Fructus aurantii obtained in this study,which is basically consistent with the literature report and field investigation.It shows that the most suitable growth of Fructus aurantii in the regional ecological environment is conducive to the accumulation of active ingredients,so as to obtain high-quality Fructus aurantii.2.The active components Naringin,Narirutin,Hesperidin,Neohesperidin were screened through the effects of monomer components on isolated animals and the compatibility of monomer components on in vivo animals,the combination of active components played a therapeutic role.The polysaccharides were extracted,isolated and purified.IR analysis showed that the polysaccharides had typical structure.GPC method showed that the molecular weight distribution was 585 k Da and 15.5 k Da KDa,GC/MS analysis mainly contained six monosaccharides(rhamnose,arabinose,xylose,mannose,glucose,galactose).The structure-activity relationship was established through the effect of Fructus aurantii polysaccharide on gastrointestinal motility in mice,which showed that Fructus aurantii polysaccharide was one of the active components of Fructus aurantii.Based on pharmacological experiments,the active components of Fructus aurantii were selected as:Naringin,Narirutin,Hesperidin,Neohesperidin and Fructus aurantii polysaccharide,and the combination of active components was more active than the monomer.3.The ecological environment of the origin influences the accumulation of secondary metabolites in the original plants of trifoliate.The typical correlation analysis of flavonoid and coumarin components in different habitats and the ecological factors of the origin shows that the ecological factors the monthly radiation in July,the average precipitation in May and the vapor pressure in April are highly correlated with Hesperidin,Tangeretin and Naringin.The results showed that the average precipitation in April and may had a high correlation with Meranzin hydrate,which indicated that there was a great correlation with the accumulation of flavonoids in Fructus aurantii,that is,the radiation in picking period,the precipitation and relative humidity in growing period,and the precipitation in growing period mainly affected the accumulation of coumarins in Fructus aurantii.The content of flavonoids,coumarins and polysaccharides in Fructus aurantii in Hunan Province is higher in general,and the composition of chemical components in different regions is different.The structure of Fructus aurantii polysaccharide showed that the number and shape of absorption peaks of Fructus aurantii in different places were different by IR.The three-dimensional morphology of polysaccharides was observed by SEM.The molecular weight distribution of polysaccharides in various regions was as follows:Crude polysaccharide from Fructus aurantii in Hunan(10.61×10~5 Da),Jiangxi(5.85×10~5 Da),Chongqing(1.88×10~5 DA)and Zhejiang(1.20×10~5 Da).The composition of monosaccharide in Fructus aurantii from different regions was basically the same,and the composition of mole ratio was different.The molars ratio of rhamnose,arabinose,xylose,mannose,glucose and galactose were Jiangxi(1.20:75.80:1.46:4.63:6.43:10.48),Hunan(0.98:73.17:10.08:2.74:2.57:10.47),Sichuan(1.44:33.38:0.87:7.07:20.37:36.87)and Zhejiang(1.72:60.37:0.36:1.00:33.43:3.13).The results showed that different components of separation and purification from different places of origin,seven components were obtained from Jiangxi,seven from Hunan,four from Sichuan and three from Zhejiang.The composition of neutral polysaccharide and acidic polysaccharide from different areas was different.The content and ratio of flavonoids,coumarins and polysaccharides,as well as the structure difference of Fructus aurantii,are affected by climate,soil and other ecological environmental factors in different places of origin.The difference of chemical components between the producing areas is the material basis of the"Geoherbalism"of traditional Chinese medicine.4.Principal component analysis and cluster analysis were used to classify the samples of Fructus aurantii from different areas.Six groups of representative samples were selected from Jiangxi Zhangshu,Jiangxi Xingan,Hunan,Sichuan,Chongqing and Zhejiang,and the compatibility ratio of active ingredient composition was fitted.The results were verified by pharmacological experiments in normal mice The results showed that the best ratio of the two groups was 1:6:1:4:20 in Jiangxi and 1:26:1:12:50 in Sichuan.5.The optimal compatibility ratio of active ingredients and their original medicinal materials have significant effects on spleen deficiency model rats.The trend of the effect of the fitting composition and its original medicinal materials on spleen deficiency model rats is similar,which indicates that the fitting composition can better reflect the efficacy of the original medicinal materials,and the drugs of each group have good therapeutic effect on model rats,improve the level of gastrointestinal hormones and regulate gastrointestinal movement.Taking the optimal matching range of active ingredients as the evaluation index,the animal experiment evaluation model was established to determine the optimal matching range of active ingredients in Fructus aurantii.6.Based on the range of optimal compatibility ratio of active components,cluster analysis was conducted to find out the samples similar to the optimal compatibility ratio results,mainly distributed in Jiangxi production area,and some of the other main production areas had"Geoherbalism"medicinal materials,indicating that the real estate area of Fructus aurantii is mainly in Jiangxi production area,and some of the other main production areas have some distribution of traditional Chinese medicine ConclusionThe results showed that the active components of the Fructus aurantii were Narirutin,Narin gin,Hesperidin,Neohesperidin and Fructus aurantii polysaccharide.The optimum compatibility range was 1:6:1:4:20 to 1:26:1:12:50.Based on the ecological environment factors,the main area s of the real estate are Jiangxi,and some of the other main production areas(Sichuan,Chongqing,Hunan and Zhejiang)have some distribution of the medicinal materials. |
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