Research Of Quality Control And Study On Anti-sepsis Mechanism Of Shengmai Injection

Shengmai injection is one of the representative medicines in the modern applications of traditional Chinese medicine.It originated from the ancient Shenginai Yin recorded in<the Theory of Internal and External Injury Debate>.In this research,the quality evaluation of Shengmai injection and the molecular mechanism of sepsis diagnosis and treatment were studied by using chemical analysis,sensitive bioactivity determination,network pharmacology and metabonomics.The experimental results were as follows:Part I:The UPLC chromatograms of 9 batches from 5 factories were analyzed by fingerprint technology,and Schisandrin chromatographic peak(No.24)was determined as the reference peak.A total of 44 common peaks were calibrated and 11 common peaks were identified,such as ginsenoside Rgl(No.10),the similarity of 9 batches of chromatograms was greater than 0.91,indicating that the difference between batches was small,the preparation process was stable,and the method was accurate and reliable.Part II:The UPLC chromatograghy method was established by quantitative analysis of multi-index components.A comprehensive evaluation model of Shengmai injection was established based on principal component analysis.The scores of each batch ranged from high to low:S2,S9,S5,S1,S6,S7,S8 and S3.Part III:The quality control standard of Shengmai injection guided by sensitive biological activity was established,which reflected its quality more objectively,clearly and comprehensively:(1)The quality control methodology based on DPPH free radical scavenging ability was established,and the scavenging ability of 9 batches of Shengmai injections from 5 factories was relatively stable;(2)The anti-inflammatory substance(NO)scavenging based on RAW264.7 cell model was proposed.Quality control methodology,closely related to its efficacy,enhance the specificity of biological activity quality control methods,9 batches of injections scavenging anti-inflammatory substance NO ability is relatively stable;Part IV:The mechanism of Shengmai injection in anti-sepsis was briefly analyzed by using network pharmacology technology.Firstly,based on the results of fingerprint analysis,9 active ingredients were confimed by Pubchem database,and about 62 possible targets of 9 ingredients were obtained from STITCH database.58 targets were collected from HPO database to meet the requirements of sepsis-related human body.STRING database was used to analyze protein interaction between chemical targets and sepsis-related targets of Shengmai injection.After deleting the unrelated targets,the interaction between drug targets and disease targets was visualized by Cytoscape v3.5.1 software.The network topological properties of nodes in the interactive network were analyzed by network analyzer plug-in,and the connectivity,median and tightness were obtained.Numbers were 19,0.014 and 0.449,respectively.The top 20 targets were selected according to connectivity ranking,the key targets in Shengmai injection network for sepsis treatment,mainly including ABL1,CCND1,CDK1,CDK2,CDK6,CDKN1B,RB1,HSP90AA1,SMARCA4,RBL2,CTNB1,MDM2,SPKRR1,LK.1.BTK,PIK3R1,TMPRSS11D,ACTG2,CD79A,RET,etc.KEGG metabolic pathway enrichment analysis obtained 20 main biological signaling pathways.It was speculated that Shengmai injection may play a synergistic role in regulating Pathways in cancer,Cell cycle,p53 signaling pathway,suggesting that Shengmai injection might participate in regulating immunity,inflammatory response,apoptosis and so on,playing a protective role in the treatment of sepsis.Specific information on the 8 major metabolic pathways was related to sepsis in KEGG analysis.Biological analysis of GO showed that the active ingredients of Shengmai injection may participate in regulating various biological processes in the treatment of sepsis,such as cell cycle,energy metabolism,intracellular signal transduction,transcriptional regulation and immune system development,suggesting that it may be used in the treatment of sepsis.It might regulate systemic inflammation and immune disorder caused by sepsis,protect secondary multiple organ dysfunction with high risk from energy metabolism,and preliminarily reveal the characteristics of multi-channel interaction regulation of the drug’s anti-sepsis pharmacodynamic activity.Part V:The mechanism of Shengmai injection on sepsis was explored by using urine metabonomics technology.The sepsis rat model was duplicated by the classical cecal ligation and perforation method.Urine was collected from different groups of rats for histological analysis.The results showed that the metabolic profile of urine showed that the metabolic distribution of rats in Shengmai injection group tended to be similar to that of rats in blank group,which proved that Shengmai injection had a good therapeutic and callback effect on sepsis rats.Among them,regulation of taurine and taurine metabolism,arginine and proline metabolism might be the key mechanisms of its treatment of sepsis.