| Objective:To make a preliminary study on the stem and leaves of Fritillaria cirrhosa and its medicinal residue,identify the chemical constituents,analyze and conduct efficacy experiment,establish its quality standard,and then develop it into veterinary medicine.Methods: 1.UPLC-MS/MS method was used to detect the chemical components in the stem,leaf and bulb of Fritillaria cirrhosa,compare the similarities between the two,and predict the efficacy of the stem and leaf of Fritillaria cirrhosa.Thirty representative chemical constituents of Fritillaria cirrhosa and Fritillaria cirrhosa on pneumonia,bronchitis and bronchial asthma were selected to study the target and mechanism of the stem and leaves of Fritillaria cirrhosa on the treatment of pneumonia,bronchitis and bronchial asthma,and their efficacy was verified by animal experiments.2.Through the establishment of alkaloid content determination method for Fritillaria cirrhosa stems and leaves,the study was conducted on the harvesting and processing methods of Fritillaria cirrhosa stems and leaves in the shade,drying at different temperatures and sun drying,to screen out the best harvesting and processing methods of Fritillaria cirrhosa stems and leaves,and conduct other inspection and identification studies to establish its quality standards.3.With alkaloid transfer rate and paste rate in the extraction process of Fritillaria cirrhosa stems and leaves as indexes,the extraction process of Fritillaria cirrhosa stems and leaves was studied by using spatial design,and the optimal extraction process of Fritillaria cirrhosa stems and leaves was selected.Then the concentration temperature and time were investigated,and the drying method was studied to optimize the optimal preparation process of the extract of Fritillaria cirrhosa stems and leaves.4.Single factor experiment was used to investigate the effect of adding excipients to Fritillaria cirrhosa stem and leaf granules,as well as different concentrations of adhesives,and orthogonal test was used to screen out the best preparation process of CBJY granules.5.The alkaloid content of CBJY granules was determined by UV-visible spectrophotometry,and the ultraviolet fingerprint and physical fingerprint were studied to establish its quality standard.6.The chemical composition of Fritillaria cirrhosa residue and bulb was detected by UPLC-MS/MS method,and the similarity between the two was compared to predict the efficacy of Fritillaria cirrhosa residue.With the guidance of network pharmacology,the target and mechanism of Fritillaria cirrhosa residue on pneumonia,bronchitis and bronchial asthma were studied.The efficacy of Fritillaria cirrhosa residue was verified by molecular docking and animal experiments.7.The use of OVA aluminum hydroxide adjuvant,the establishment of experimental rat asthma model,and then according to the ammonia induced cough experiment,serum inflammatory factors content and other pharmacodynamic indicators,as well as the comparison of lung tissue pathological sections,to investigate the effect of Fritillaria cirrhosa stem and leaf extract on rats.8.By studying the contents of water,organic matter,nitrogen,phosphorus and potassium in the stem and leaf residues of this shell,the possibility of its fermentation as bioferfertilizer was explored,and the effects of adding EM bacterioides and compound bacterioides on its fermentation effect were studied.Results: The chemical composition similarity between stem and leaf of Fritillaria cirrhosa and bulb reached 90.2%.The stem and leaf of Fritillaria cirrhosa exhibited antitussive and antisputum effects on pneumonia,bronchitis and bronchial asthma mainly through CCND1,EP300,CDK2 and other targets.There was no significant difference in the number of coughs between stem and leaf water extraction group and codeine group and acute branch syrup group,but both groups could significantly increase the amount of phenol red excretion.2.The quality standard of Fritillaria cirrhosa stems and leaves was established.The total alkaloids in Fritillaria cirrhosa stems and leaves were not less than 0.156%,and Fritillaria cirrhosa stems and leaves were not less than 0.00128%.3.The extraction process of the stem and leaves of Phyllotillate tuber was selected as the concentration of A alcohol(46%～58%),extraction time of B(70min～174min),and extraction times of C(1-3 times).The concentration conditions were 60℃ within 8h and the drying method was vacuum drying at60℃.4.The optimum preparation technology of CBJY granules was as follows: dry paste powder and auxiliary material ratio of 4∶2,ethanol concentration of 85%,mesh number of 14.5.The quality standard of CBJY particle was established.The similarity limit of fingerprint of CBJY particle was 0.795,and the similarity limit of UV fingerprint was 0.893.6.The overlap rate of compounds between the residues and the bulbs reached 83.6%.Three main chemical components,91 corresponding targets and 21 therapeutic pathways were selected by network pharmacology,and the binding energy of core compounds was lower than that of chemical agents.The incubation period and cough frequency of mice were significantly prolonged and decreased,respectively,and the swelling of mice ear was significantly inhibited.7.The cough incubation period of rats in stem and leaf group of Fritillaria cirrhosa group was similar to that in the model group,the content of TNF-α was significantly reduced.Pathological observation results the alveolar deformation of stem and leaf group of Fritillaria cirrhosa group showed an obvious recovery trend,some of which had been recovered,and inflammatory cell infiltration was also reduced.8.The average organic matter content of CBJY residues was 88.23%,and the total nutrient content was 1.80%.The total nutrient content of bio-organic fertilizer of the residues of stem and leaves of Fritillaria cirrhosa increased after adding complex bacteria.Conclusion:The stem,leaf and residue of Fritillaria cirrhosa have high chemical composition similarity with the bulb of Fritillaria cirrhosa,and both have the effect of relieving cough and reducing sputum.The alkaloid content of the stem and leaf of Fritillaria cirrhosa is higher than that of the bulb.The quality standard of the stem and leaf of Fritillaria cirrhosa was established,the best preparation process of the stem and leaf extract of Fritillaria cirrhosa was selected,the quality standard of CBJY granule was established.The stem and leaf extract of Fritillaria cirrhosa can inhibit the cough of asthmatic-model rats,reduce the deformation of alveoli of rats to a certain extent,and reduce the level of inflammatory factors.The CBJY residue can be used as raw material for the fermentation of bio-organic fertilizer,and the compound bacteria is helpful for the fermentation of bio-organic fertilizer from the stem and leaf of Fritillaria cirrhosa. |
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