Studies On Preparation,quality Standard And Potential Pharmacodynamic Of Sanren Granules

Objective: In this paper,the extraction and forming process of Sanren Granules was optimized,the relative critical humidity of granules was determined.The quality standard and fingerprint of Sanren Granules were established,and markers of quality difference were screened.The model of damp-heat gastritis in rats was established based on network pharmacology,and the effect of Sanren Granules on damp-heat gastritis in rats was studied,provided theoretical basis for further development and research of Sanren Granules.Methods:(1)The prescription of Sanren Decoction was verified through literature review.According to the records in ancient books,combined with modern practice,the extraction process of Sanren Decoction was optimized by comparing the standard decoction of ancient method and orthogonal test with the amount of paste,the content of amygdalin and 50% ethanol extract as evaluation indexes.The forming rate,angle of repose,bulk density and moisture absorption rate were used as evaluation indexes,the forming process of Sanren Granules was optimized by using Central Composite Design-Response Surface Method.(2)The quality standard of Sanren Granules was established by determining the general inspection,thin layer identification and content determination method of granules.(3)The fingerprint of ten batches of Sanren Granules was established by HPLC.Cluster analysis,orthogonal least squares-discriminant analysis(OPLS-DA)and variable importance projection(VIP)values were used to screen the quality difference markers of Sanren Granules.(4)TCMSP was used to screen the chemical components of Sanren Granules.The gene library search function was used to obtain the relevant target information for the symptoms of damp-heat gastritis.The intersection of drug and disease-related target genes was taken.The STRING database was used to construct the PPI network for the selected main targets.The corresponding network diagram was constructed by Cytoscape software.The DAVID database was used to analyze the GO enrichment analysis and KEGG enrichment analysis of Sanren Granules for the treatment of damp-heat gastritis related target genes.The rat model of damp-heat gastritis was established.The differences in body weight,body temperature and fecal water content of rats before and after administration in different administration groups were compared.Enzyme-linked immunosorbent assay(ELISA)was used to determine the contents of IL-1β(Interleukin-1β),IL-6(Interleukin-6),TNF-α(Tumor necrosis factor-α),HSP-60(Human heat Shock protein-60)and HSP-70(Human heat Shock protein-70)in serum.Results:(1)The best extraction process of Sanren Decoction was as follows: soaking for one hour with 9.5 times the amount of water,boiling for one hour,and then boiling for one hour with 8 times the amount of water.The optimum molding process of Sanren Granules was as follows: dextrin was used as excipient,the ratio of drug to excipient was 1:1.5,the volume fraction of ethanol was 78%,the amount of ethanol was 0.21 times of the total amount of drug powder and excipient,and the relative critical humidity of Sanren Granules was 74.45%.(2)The general investigation index of Sanren Granules and the thin layer identification methods of Bitter almond,Semon coicis and Magnolia officinalis were established,the content determination method of amygdalin was established.(3)A total of15 common peaks were identified in the fingerprints of 10 batches of samples,and the similarity was greater than 0.90.The cluster analysis showed that the 10 batches of samples could be divided into four categories,S1 and S9 as one category,S7 as one category,S2 and S8 as one category,S3-S6 and S10 as one category.The results of OPLS-DA and VIP values showed that the VIP values of No.8 peak,No.9 peak(amygdalin)and No.10 peak were all greater than 1.(4)The relevant active ingredients in Sanren Granules were identified by constructing a composition-target network,and 44 targets for the treatment of dampness and heat were identified by matching drug targets and disease targets,and the five most relevant targets TP53,IL-6,JUN,IL-1β and VEGFA were found through the protein interaction network.GO and KEGG enrichment analysis showed that Sanren Granules inhibited inflammation,regulated hormone secretion and treated damp-heat gastritis through multi-protein and multi-signaling pathways.The contents of IL-1β,IL-6 and TNF-α in the serum of rats with damp-heat gastritis model were significantly higher than those in the control group(P<0.01),and were significantly decreased after treatment with medium and high doses of Sanren Granules(P<0.01).Compared with the control group,the contents of HSP-60 and HSP-70 in the serum of the model group were significantly increased(P<0.01).After treatment,compared with the model group,the contents of HSP-60 and HSP-70 in each administration group were significantly decreased(P<0.01).Conclusion: The extraction rate of active components was high,stable and reliable.The molding process had the advantages of high molding rate,good fluidity and simple process.The quality standard and fingerprint examination items are stable and reliable,which can provide a theoretical basis for the quality control of Sanren Granules.No.8 peak,No.9 peak(amygdalin)and No.10 peak may be the differential markers affecting the quality of Sanren Granules.The model of damp-heat gastritis in rats was established by network pharmacology,and the difference of efficacy of Sanren Granules n in treating damp-heat gastritis in rats was clarified,which provided theoretical basis for the basic research and clinical application of Sanren Granules.