Exploring The Compatibility Rules Of Active Components Of Traditional Chinese Medicine Based On LPS-induced Cellular Inflammation Model

The multi-channel and multi-target treatment of traditional Chinese medicine prescription is a typical example of drug combination,which shows great advantages in the treatment of diseases.The research on the compatibility relationship of traditional Chinese medicines mainly includes the compatibility of Chinese herbal medicines,the compatibility of effective parts and the compatibility of active ingredients.The compatibility between efficacy-oriented active ingredients combines traditional experience with modern technology,it can not only exert the effect of synergistic treatment,but also elucidate the mechanism of action at the molecular level,which provide a new idea for the traditional Chinese medicine modernization.The development of many diseases(such as cancer,infectious diseases)will be accompanied by the production of inflammation,the nine herbs,Bupleuri Radix,Citri Reticulatae Pericarpium,Paeoniae Radix Rubra,Zingiberis Rhizoma Recens,Glycyrrhizae Radix Et Rhizoma,Arctii Fructus,Magnoliae Officinalis Cortex,Forsythiae Fructus and Lonicerae Japonicae Flos,have significant curative effects in anti-inflammatory and anti-virus.The 9 herbs are all mentioned in the recommended Chinese herbal formulas for the treatment of COVID-19,and have played a positive role in relieving symptoms,preventing complications and improving the cure rate.we established LPS-induced RAW264.7 cells as an inflammation model for screening anti-inflammatory active drugs,and explore the compatibility and interaction of the active ingredients of nine traditional Chinese medicine.The research content is mainly divided into the following three parts:1.We selected 12 anti-inflammatory active compounds in 9 medicinal materials,the effects of 12 compounds on the viability of RAW264.7 cell was explored by MTT,and the safe concentration ranges of the compounds were determined;The Griess method was used to explore the inhibitory effect of the drug on the release of NO in the LPS-induced cell supernatant.Among them,10 compounds with anti-inflammatory activity were screened out,namely saikosaponin a,saikosaponin b2,nobiletin,paeoniflorin,6-gingerol,glycyrrhizic acid,arctigenin,honokiol,forsythoside A and luteolin.2.Drug-pair compatibility studies of anti-inflammatory active compounds.Based on the compatibility research strategy of standard substances with clear pharmacological effects,the pairwise compatibility of 9 compounds produced 36 combinations.The antiinflammatory activities of the 36 combinations were evaluated by Griess,and the drug pair compatibility interactions was explored by the drug synergy evaluation model of "Chou-Talalay".Through high-throughput screening,there are 17 groups of drug pairs that produce synergistic effects(CI<1),among them,nobiletin,saikosaponin b2 and paeoniflorin appeared more frequently,and had the potential to produce synergistic effects in combination with other drugs.We also determined a group of the best compatible drug pair and its best ratio is saikosaponin b2: glycyrrhizic acid = 1: 10.3.A multi-component compatibility study based on traditional Chinese medicine prescription.First,we studied the anti-inflammatory activity of M8,which was composed of 8 compounds that produced synergistic effects in the drug-pair compatibility.It was observed that M8 inhibited the cell normal growth at two high concentrations,but at the other three compatible concentrations,the NO content decreased in a concentrationdependent manner and no obvious changes in cell state.Then,based on the compatibility strategy of the active ingredients of “traditional Chinese medicine prescription”,the 8compounds were combined in three-three,four-four and five-five combinations,and evaluate the anti-inflammatory activities and compatibility interactions of the obtained 13 drug combinations.Finally,10 groups of drug formulations with synergistic antiinflammatory properties were screened,7 of them had better synergistic effect,these synergistic anti-inflammatory better combinations can be used as candidate formulations of anti-inflammatory drugs,and obtained a group of the best compatibility formula C2 and its best ratio is saikosaponin b2: nobiletin: glycyrrhizic acid: honokiol: forsythoside A=10: 4: 140: 1: 30.