| BackgroundA?aí is a commonly used herb in South America and has a long application history.Modern studies have shown that A?aí is rich in anthocyanins and other flavonoids,and has various activities such as anti-oxidation,anti-inflammatory and anti-cancer activities.As an international hotspot plant,A?aí is widely used.After years of hard work,the research program successfully introduced A?aí into China in 2013.At present,there are a variety of products that are compatible with traditional Chinese medicine in China,but due to the lack of traditional Chinese medicinal properties and clinical standards of Chinese medicine,their applications are varied,and the compatibility is abnormally disordered.Therefore,it is particularly urgent to study the TCM property of A?aí.Based on the research progress of A?aí,the research program has conducted a comprehensive review of its literatures using database technology.We conducted a theoretical analysis of A?aí’s TCM properties and taking cold and hot properties as the entry point,the theoretical research results of the cool nature of A?aí were taken as the scientific hypothesis,and verified that A?aí’s TCM properties is cool Pharmacologically.In the next study,Starting from the basic point of view to carry out further study of A?aí’s cool nature.In recent years,a variety of new models have emerged in the research of medicinal properties.Among them,Prof.Kuang Haixue creatively proposed a research model on the theory of TCM property and flavor based on TCM splittability,and considered that the Chinese herbal medicine should be split in all components through a variety of techniques in combination with each other.Separation of components does not cross-research and the chemical composition does not change,and study with the animal experiments to determine the material basis to play a related role,so as to summarize the property and flavor attribution of the separated components,and finally return to Chinese medicine theory.Based on the study of properties of A?aí in the past research,this progrom will further study the chemical composition combined with the theory of TCM property and flavor.Objective and significanceThe research on the medicinal properties of TCM is inseparable with the material basis.Based on the literature research,theoretical discussion and verification of the medicinal properties of A?aí,this program splited A?aí into A?aí oil,A?aí alcohol extract,A?aí water extract and had a composition analysis,and finally combined with the animal experiment to clarify the material basis of the cold and cool drug properties of the A?ai.This research provides a reference for the study of the medicinal resources of traditional Chinese medicine,and explores a new method for the basic research of material basis,and provides experimental basis for the application and development of A?aí,and laying a theory and experimental basis for combining application of A?aí with Chinese herbal resources.MethodUnder the guidance of the theories of TCM,this article has studied the cold and hot medicinal properties of A?aí from the aspects of pharmacological research and pharmacological research combined with the theory of TCM property and flavor based on TCM splittability.The details are as follows:Pharmaceutical research:1.A?aí’s chemical composition study:The cyanidin-3-O-glucoside,total flavonoids and polysaccharides were determined by HPLC and ultraviolet spectrophotometry.2.The establishment of the A?aí split fraction:firstly,the yield of oil was used as the index,and the effect of extraction pressure,extraction temperature,and extraction time on the yield of oil was examined by a single factor experiment,and the best conditions for extracting A?aí oil by supercritical CO2 fluid was further determined by orthogonal experiments.Secondly,the transfer rate of cyanidin-3-O-glucoside and total flavonoids was used as the index,and the single-factor experiment and orthogonal experiment were used to inspect the best process.The extraction is performed,and the best process is preferred.Finally,the polysaccharide transfer rate as an index,the single-factor experiment and orthogonal experiment were used to establish the preparation process of A?ai water extract.3.Chemical composition study of the A?aí split fraction:GC-MS was used to analysis A?aíoil after methyl esterification and HPLC was used to determine β-sitosterol in A?aí oil.The content of the HPLC fingerprint of the A?aí alcoholic extract was established,and the content of cyanidin-3-O-glucoside was determined.The content of total flavonoids was determined by ultraviolet spectrophotometry.The content of polysaccharide in the water extract by ultraviolet spectrophotometry.Finally,qualitative analysis were used to study the mutual non-intersection of the A?aí split fraction.Pharmacology research:1.Biological effects of A?aí on the Deficiency-cold Syndrome and Deficiency-hot Syndrome mice:After adaptive feeding for a period of time,KM mice were randomLy divided into blank group,Deficiency-hot group,Deficiency-cold,Deficiency-hot+Rehmannia group,Deficiency-hot+A?aí group,Deficiency-cold+Cinnamomi Cortex group,Deficiency-cold+A?aí group;the mice were administered orally at the dose of 160mg/kg thyroxine and 25mg/kg hydrocortisone on the afternoon from the 8th day per day respectively for 14 days to induce Kidney-Yin and Kidney-Yang models;the changes of temperature trend of mice were investigated by cold/hot plate differentiating assay;the levels of cAMP,cGMP,cAMP/cGMP in serum were detected in enzyme-free method,and the T3 and T4 were detected by radioimmunoassay to investigate the trend of A?aí.2.Biological effects of A?aí split fraction on the Deficiency-cold Syndrome and Deficiency-hot Syndrome mice:After adaptive feeding for a period of time,KM mice were randomLy divided into blank group,Deficiency-hot model group,Deficiency-hot+A?aí group,Deficiency-hot+A?aí oil group,Deficiency-hot+A?aí alcohol extract group,Deficiency-hot+A?aí water extract group,Deficiency-cold model group,Deficiency-cold+Cinnamon group,Deficiency-cold+A?aí group,Deficiency-cold+A?aí oil group,Deficiency-cold+A?aíalcohol extract group,Deficiency-cold+A?aí water extract group.The mice were administered orally at the dose of 160mg/kg thyroxine and 25mg/kg hydrocortisone on the afternoon from the first day per day respectively for 14 days to induce Kidney-Yin and Kidney-Yang models.The changes of temperature trend of mice were investigated by cold/hot plate differentiating assay;the levels of cyclic nucleotides,thyroid hormones,immunity,endocrine and metabolism levels were determined to clarify the material basis of the cool drug properties of the A?aí.ResultPharmaceutical research:1.A?aí’s chemical composition study:A?aí contains cyanidin-3-O-glucoside 0.142mg/g,total flavonoids 41.61mg/g,polysaccharides 28.40mg/g.2.The establishment of the A?aí split fraction:(1)A?aí oil:The oil yield was used as the index to determine the optimum process for the extraction of A?aí oil in supercritical CO2,and the extraction pressure is 25 MPa,extraction temperature is 45℃,extraction time is 90 Min,the oil yield reaches 53.645%.(2)A?aí alcohol extract:The total flavonoid and cyanidin-3-O glucoside extraction rate transfer rate were taken as the indicators.Comprehensive consideration was given to the final determination of the ethanol extract process with 10 times 80%ethanol and ultrasonic extraction 3 times for 1 hour each time.(3)A?aí water extract:Orthogonal experiment optimizes the process of A?aí water extract and the result is adding the ratio of material to liquid 1:6,extracting time is 3 times,extraction time is 1h,and the polysaccharide transfer rate is 91.25%.3.Chemical composition study of the A?aí split fraction:(1)A?aí oil:34 compounds were identified and identified from A?aí oil,among which oleic acid,palmitic acid,linoleic acid,palmitoleic acid,stearic acid are the most;the mass fraction of β-sitosterol in the A?aí oil is 1.83 mg/g.(2)A?aí alcohol extract:A?aí alcohol extract were studied by HPLC.The content of cyanidin-3-O-glucoside was calculated to be 0.488 mg/g in solids,and the total flavonoid content was 192.10 mg/g in solids.(3)A?aí water extract:A?aí water extract contained 381.27 mg/g polysaccharide.Pharmacology research1.Biological effects of A?aí on the Deficiency-cold Syndrome and Deficiency-hot Syndrome mice:Two weeks after the model was established,the deficient-hot syndrome mice showed symptoms such as dry hard stools,increased voluntary activities,and easy irritability.The mice showed signs of weight loss,loss of luster in the hair,unresponsiveness,and listlessness,confirming that the model was successful.After giving drugs,both the Rehmannia group and the cinnamon group improved their daily status,ethology,and biochemical indicators to some extent.The A?aí is consistent with the Rehmannia,and it has different degrees of treatment for the Deficiency-hot Syndrome mice,but contrary to the cinnamon,the effect on the Deficiency-cold Syndrome mice is not obvious.This experiment verifies the cool properties of A?ai from the point of view of comparing similar types of different kinds of anti-evidence.2.Biological effects of A?aí split fraction on the Deficiency-cold Syndrome and Deficiency-hot Syndrome mice:we studied the effect of A?aí split fraction in Deficiency-cold Syndrome and Deficiency-heat Syndrome Mice in Temperature Tendency,Cyclic Nucleosides,thyroid hormones,immunity,endocrine,and metabolism levels,and ultimately determined A?aí is cool,and A?aí alcohol extract are the material basis of the cool drug properties.The A?aí oil and A?aí water extract are warm,A?aí is the result of the combined action of them.ConclusionsA?aí has certain improvement on Deficiency-hot Syndrome mice,and it has no obvious effect on Deficiency-cold deficiency Syndrome mice,indicating that A?aí’s cool.Preparation Process of A?aí split fraction A?aí oil,A?aí alcohol extract and A?aí water extract were established through the single-factor experiment and orthogonal experiment.The preparation process was stable and feasible,and the chemical components of each split fraction was simultaneously determined.And they do not cross each other.By examining the effects of A?aísplit fraction on the Deficiency-cold Syndrome and Deficiency-hot Syndrome mice,it was shown that the alcoholic properties of A?aí extracts were cool,while the A?aí oil A?aí water extract were warm.Finally,the medicinal propertie of A?aí is the result of the combined effects of the split fractions.A?aí is cool,and A?aí alcohol extract are the material basis of the cool drug properties.It is because of the combination of cold components and warm components in the A?aí,the overall appearance of A?aí is cool,and from the perspective of chemical composition explains the reason of the cool but not cold of the A?aí. |
PDF Full Text Request