| Purpose: Network pharmacology-Analysis of the target of drug co-action components explores the pharmacological effects of Qingrun granules at the molecular biology level; analyzes and explores the best extraction technology of traditional Chinese medicine granules,Qingrun granules,and the best preparation technology of Qingrun granules raw powder;for Qingrun The pharmacological effects of the granules for relieving cough,reducing phlegm,anti-inflammatory,and immune regulation are verified by pharmacodynamics,providing basic experimental basis for clinical research.Materials and methods: 1.Use modern network pharmacology experimental database to explore the molecular-level pharmacodynamic mechanism of the main drug components of Qingrun Granules.2.Using orthogonal design,based on the traditional prescription principles of the monarch and the minister and the modern pharmacological research content,it can be used to treat honey Stemone,bitter almond,lily,Ophiopogon japonicus,Adenophora glabra,Fritillaria,Mulberry leaf,Thyme grass,Licorice The main pharmacological components of the drug are heated,condensed and refluxed,evaporated and concentrated and dried to make a dry paste.The dry paste is ground and crushed,and the powder is sieved through 80 mesh pharmacopoeia.The four main components are phytosterine and amygdalin.,Ophiopogon saponins D,and ammonium glycyrrhizinate were analyzed by HPLC high performance liquid phase.Based on the results of the component analysis and the yield rate,the AHP analytic hierarchy weighted score calculation was performed to optimize the best extraction process.3.On the basis of the best extraction process,according to the excipient process guidelines and requirements of the 2020 Chinese Pharmacopoeia Sibu Granules,the excipient process research of Qingrun Granules is carried out.The ratio of graduate drug to dextrin is 1:1.The difference in molding rate,moisture absorption rate,and solubility under different drug-adjuvant ratios of 1:2,1:3,and 1:4,so as to obtain the best shaping process for Qingrun granules in the laboratory.4.To verify the pharmacodynamics of cough relieving,phlegm resolving,anti-inflammatory,and immunomodulation on the raw powder obtained from the 2best extraction processes,96 SPF BABL/c mice,4 to 6 weeks old,were randomly divided into Qingrun The antitussive group,the phlegm-resolving group,the anti-inflammatory group,and the immunomodulation group were each divided into a moisturizing group,a western medicine group,and a blank group according to the mode of administration.In the antitussive group,the cough induced by ammonia water was used to induce cough in each group of mice in a glass cylinder saturated with concentrated ammonia water,and the incubation period of cough induction and the number of coughs within five minutes were recorded;the phlegm-reducing group was used the phenol red excretion method,and the mice were injected intraperitoneally with phenol red Solution,after a period of time,peel off the complete isometric trachea and place it in sodium bicarbonate solution,centrifuge and shake to ensure that the phenol red in the trachea is completely released.After centrifugation,the supernatant will be used as a blank,zeroed with distilled water as a blank,and measured under a UV spectrophotometer.The OD values ??of the supernatants of each group were separately weighed and a certain amount of phenol red was diluted by multiples to obtain a standard solution of phenol red sodium bicarbonate with an equal concentration gradient,and the OD value was measured.The OD value is the ordinate and the phenol red concentration is the abscissa.Establish a fitting function,calculate the phenol red content according to the standard curve equation and the supernatant OD value,calculate the corrected phenol red content according to the weight of the mouse,and compare the phlegm reduction rate.In the anti-inflammatory group,xylene was applied to the left ear auricle of the mouse to cause inflammation,and a 6mm round ear piece was punched on the same part of the left and right ears with a punch,weighed,and the difference was calculated.The immunomodulatory group performed a serum hemolysin test.After SRBC sensitization,blood was taken from the orbit of the mouse,and the serum was obtained by centrifugation.The anti-sheep red blood cell antibody in the serum was analyzed by hemagglutination qualitative analysis and antibody OD value detection method.Result:1.The target genes most closely related to the results of network visualization and the drug action of Qingrun granules are CDRT15L2,PGLYRP-4,STYXL12.The best extraction process: The best extraction process for Qingrun granules is 8 times70% ethanol as the solvent,heating and condensing reflux method for 3 hours,3 times extraction.3.The best shaping process: the ratio of Qingrun granular raw powder to dextrin is 1:3,85% ethanol is used as a binder,and wet granulation.4.Validation results of pharmacodynamic experiments: Anti-cough group: Qingrun granules can prolong the incubation period of cough in mice and improve the tolerance of the mouse’s respiratory tract to the stimulation of concentrated ammonia;reduce the number of coughs stimulated by concentrated ammonia,which is caused by stimulation of concentrated ammonia The stress cough of mice is time-dependent,and there are obvious differences between the three groups within 20s～180s.Phlegm-resolving group: The phlegm-reducing rate of each experimental group: Qingrun group=121.995%,Western medicine group=140.497%,blank group=100%,Qingrun granules can promote the discharge of sputum in the trachea and increase the phlegm-reducing rate.Anti-inflammatory group: The weight difference between left and right ears: The difference between the weight of left and right ears in the experimental group and the blank group was reduced,and the difference was statistically significant(P<0.05).Immunomodulatory group: The amount of antibody produced by serum hemolysin test in the Western medicine group was greater than that in the Qingrun group,and the difference was statistically significant(P<0.05),and the Qingrun group was greater than the blank group,and the difference was statistically significant(P<0.05).Serum hemolysin agglutination test: Dexamethasone group﹥Qingrun granule group﹥normal saline group.in conclusion:1.The composition-target network visualization of Qingrun granule compound medicine can explain the internal mechanism of drug action from the perspective of molecular biology,and use modern information technology to provide ideas for new drug research and development.2.The weighted scoring method of AHP analytic hierarchy process is conducive to the optimization of the extraction of active ingredients of Chinese herbal medicines,and it is convenient for the research of Chinese herbal medicine.3.In the exploration of the wet granulation and shaping process in the laboratory,the factors that affect the molding process include the types of excipients,the ratio of auxiliary materials,the types of binders,and the choice of additives,but the most critical factors affecting the molding are the raw powder and auxiliary materials.Proportion provides a reference for the exploration of the shaping process of mass production in pharmaceutical factories.4.The strength of Qingrun granule’s cough,phlegm,anti-inflammatory,and immunomodulatory effects is verified.Based on the experimental procedures and procedures and analysis methods of the antitussive effect,it lays a foundation for verifying the pharmacodynamic effects of Qingrun Granules’ antiasthmatic effects. |
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