Study On The Extraction Process And Anti-inflammatory Activity Of The Main Active Components Of Yifei Jisheng Prescription

BackgroundChronic obstructive pulmonary disease(COPD)is a disease that features incompletely reversible airflow obstruction and regarded as a disease of significant morbidity and complex pathogenesis.The inflammatory response is the most important pathological feature of COPD,so suppression of the inflammatory plays a key role in the mitigation and treatment measures for COPD.Yifei Jisheng prescription has a very significant therapeutic effect in treating for stable COPD.The article researched and explored the anti-inflammatory effect and mechanism of major components of Yifei Jisheng prescription.The extraction technics of Yifei Jisheng prescription was investigated in this article.The water soluble ingredients in the radix astragali were extracted by water decocting method,and other medical materials were extracted by alcohol refluxing extraction to screen out the optimal extraction process.The paper researched the anti-inflammatory effect and mechanism of major components of Yifei Jisheng prescription by using the optimal concentration of tumornecrosis factor-?(TNF-?)to stimulate A549 cells to establish a model of COPD imflammatory.ObjectiveTo optimize the water decocting and alcohol refluxing extraction technique conditions according to the main components of Yifei Jisheng prescription;To research and explore the anti-inflammatory effect and mechanism of major components of Yifei Jisheng prescription by establishing a model of COPD imflammatory and to further clarify the molecular mechanisms for the treatment of COPD by observing the changes of cytokines such as IL-8?MMP-9?TNF-?.Method 1 Study on extraction process of Yifei Jisheng prescriptionAccroding to the different components of Yifei Jisheng prescription,water extraction and ethanol refluxing extraction were carried out respectively.The water extraction technologies were optimized by orthogonal experiment and multiindex compositive score method with the amount of water,extraction times and extraction time as factors using the extraction transfer rate of astragaloside IV,paeoniflorin and bergenin as index.The ethanol extraction technologies were optimized with ethanol concentration,amount of ethanol,extraction time and extraction times as factors using the extraction transfer rate of loganin and deoxyschizandrin as index.2 Effects of active components in Yifei Jisheng prescription on inflammation model of A549 cellsBy establishing a model of COPD imflammatory using the optimal concentration of tumornecrosis factor-?(TNF-?)to stimulate A549 cells,the concentration ranges of the major components were selected by MTT method;the experiments were divided into blank group(the cells are cultured by complete medium),model group(the cells are cultured by complete medium containing TNF-?)and experimental group(the cells are cultured by different concentration of components of Yifei Jisheng prescription),the secretion and changes of cytokines such as IL-8?MMP-9?TNF-? are observed and measured by enzyme linked immuno sorbent assay(ELISA).Result 1 Study on extraction process of Yifei Jisheng prescriptionOptimal water extraction technology of Yifei Jisheng prescription was as follows:extracting twice with 12 folds of water for 1h each time.And the optimal ethanol refluxing extraction technology was extracting for 2 times and 1h each time with 10 folds amount of alcohol of 70%.Combined water extraction and alcohol extraction concentrate,and then dried by vacuum drying method.2 Effects of active components in Yifei Jisheng prescription on inflammation model of A549 cellsThe bergenin can significantly inhibit the secretion of TNF-?(P<0.01);the quercetin can significantly inhibit the secretion of IL-8(P<0.01),and inhibit the secretion of MMP-9 and TNF-?(P<0.05);the kaempferol can significantly inhibit the secretion of IL-8?MMP-9 and TNF-?(P<0.01);the schizandrin B can significan-tl y inhibit the secretion of IL-8?MMP-9(P<0.01),and increase the secretion of TI MP-1 and MMP-9 in the meanwhile(P<0.01);the nobiletin have different deg-ree s of inhibition on the secretion of IL-8?MMP-9 and TNF-?;the synephrine ca n inhibit the secretion of IL-8(P<0.05),and MMP-9 and TNF-? significantly(P<0.01);the hesperidin can can inhibit the secretion of MMP-9(P<0.05)and IL-8(P<0.01)effectively;the betaine can inhibit the secretion of MMP-9 and TNF-?(P<0.01);the lycium barbarum polysaccharides can inhibit the secretion of TNF-?(P<0.05);the verticine can inhibit the secretion of IL-8(P<0.01)?MMP-9(P<0.05)and T NF-?(P<0.05);and the verticinone can significantly inhibit the secretion ofTNF-?(P<0.01);both of thepaeoniflorin and paeonol can inhibit the secretion of MMP-9(P<0.01);the astragalus polysaccharide can inhibit the secretion of MMP-9(P<0.05)and TNF-?(P<0.01)in a certain degree.Conclusion 1 The optimum extraction technology is simple with a stable and reliable process,the method will increase yield of the main active components of Yifei Jisheng prescription.It also forms a base for manufacture process in new drug development.2 Some effective components of Yifei Jisheng prescription,such as bergenin,quercetin,kaempferol,schizandrinB,nobiletin,synephrine,hesperidin,verticine,icariin and astragalus polysaccharide have significant inhibitory effect on the secretion of inflammatory cytokines such as IL-8?MMP-9 and TNF-?,and have significa-nt anti-inflammatory actions,which has laid a foundation for further studies of components compatibility in Yifei Jisheng prescription.