Study On The Preventive Effect And Active Components Of Red Peony Root On Endotoxin-induced Acute Lung Injur

Acute lung injury(ALI)is a common clinical inflammation,the main features are respiratory rate changes and respiratory distress,non-cardiogenic pulmonary edema and refractory hypoxemia,caused by direct,indirect injury factors.SARS,H5N1,H7N9 and other respiratory diseases are leading to varying degrees of ALI,a serious threat to human life and health.ALI belongs to the category of science of epidemic febrile disease of traditional Chinese medicine,the study found that the pathogenesis of the disease may be excessive uncontrolled inflammatory reaction.For the main pathogenesis,the main clinical use of detoxification,blood circulation,Qi and other treatment methods.The rats were treated with Qingre Liangxue Decoction to induce acute lung injury induced by intratracheal endotoxin in rats.The preventive effects of Radix Paeoniae Albra on acute lung injury were studied and the characteristic map of red peony was prepared.The relationship between HPLC characterization and pharmacodynamics was studied by regression analysis.Lay the foundation for the study of pharmacological material basis,with a view to future service in clinical.This paper mainly studies and discusses from the following five chapters:The first chapter is the literature review part,this chapter is mainly on the chemical composition,pharmacological effects,fingerprints and acute lung injury research,mechanism,the relationship between cytokines and acute lung injury,the progress of drug treatment and TCM syndrome differentiation.According to the literature research,Radix Paeoniae Rubra in the anti-LPS,anti-inflammatory aspects of a certain therapeutic effect on the red peony intervention in acute lung injury spectrum of the relationship between the study,to help its material basis for further mining research,can be more comprehensive.The evaluation of the treatment of Radix Paeoniae Rubra,to guide the clinical rational and safe medication.The second chapter is the research ideas and the main contents of this subject,through the first chapter of the literature research to determine the subject of research content and technical route.The third chapter is the effect of Radix Paeoniae Rubra on LPS-induced acute lung injury in rats,including endotoxin-induced acute lung injury in rats and the preventive effect of Radix Paeoniae Albra on endotoxin-induced acute lung injury in rats.The first section LPS were injected into the endotracheal tube(LPS),and the animals were sacrificed at different time points after modeling.The number of white blood cells,the protein concentration,the number of inflammatory cells and the wet/dry ratio(W/D)of the upper right lobe were detected in the alveolar lavage fluid(BALF).The results showed that the number of white blood cells in BALF was significantly increased(P<0.01),the protein concentration and W/D were also increased(P<0.01),the number of white blood cells reached the peak at 24 h,the protein concentration and W/D reached its peak at 12h.Under the microscope,the number of inflammatory cells in the lung tissue and Gimsa staining were observed under the microscope.It was found that the integrity of the alveolar structure was destroyed after the model,and the infiltration of a large number of inflammatory cells was observed.Through the dynamic study,the final selection of 12h for the extraction time,and detection of white blood cell count,protein concentration,lung wet and dry than the indicators,and observe the pathological status of lung tissue.The second section after the establishment of the model,the effects of different doses of Radix Paeoniae Rubra on the prevention of acute lung injury in LPS on rats were studied.The positive drug was treated with dexamethasone.The results showed that the levels of WBC(P<0.05)and protein concentration(P<0.05)were significantly decreased in the dexamethasone group,high dose and middle dose groups(P<0.05),relieve pulmonary edema,alveolar structure damage,inflammatory cell infiltration,improve lung bleeding.In the middle dose group,the four indexes were the closest to the positive drug group.The above indexes and the optimal dose were the best dose,which had obvious effect on the prevention of acute lung injury.The fourth chapter is the establishment of liquid chromatogram and the LC/MS analysis.The chemical constituents corresponding to the chromatographic peaks of Radix Paeoniae Rubra were analyzed by HPLC-MS/MS,and the attribution of the chromatographic peaks was determined according to the related literature.The extraction of LC-MS was carried out in the extracts of water extract,30%ethanol extract,50%ethanol extract,70%ethanol extract,90%ethanol extract and water extract Fingerprint studies to obtain fingerprint information.LPS-induced acute lung injury model was used to determine the inhibitory protein,WBC and lung W/D activity.The characteristic spectra of the six extracts(21 chromatograms)were combined with pharmacological experimental data by partial least squares.The regression coefficients of the dependent variable Xn(the peak area on the fingerprint)with respect to the dependent variable Yn(inhibition rate of protein,leukocyte and lung W/D)were obtained by spectral correlation analysis.Based on the percentage of the product of the positive regression coefficient and its corresponding peak area as the contribution rate of the drug to determine the combination of the peaks with the greatest contribution to the efficacy,thereby determining the functional ingredient group.In the spectral analysis results,the results showed that X1,X11 and X12 were the main active components of Radix Paeoniae Rubra on acute lung injury,and the contribution rate was over 80%.The contents of X1,X11 and X12 were gallic acid,paeoniflorin and ethyl gallate respectively by comparison of reference substance and HPLC-MS/MS.Chapter 5 is the separation and identification of functional components.The extracts were extracted by macroporous resin,silica gel column chromatography and gel.The functional components were identified by high performance liquid chromatography(HPLC)and thin layer chromatography(TLC).Six compounds were isolated:X1(paeonol),X11(paeoniflorin),X12(ethyl lactate),X18(benzoyl paeoniflorin),X5(paeoniflorin),X20(paeonol).In this study,we have studied and validated the preventive effect of Radix Paeoniae Albra on ALI model,established the characteristic Spectrum of Radix Paeoniae Rubra extract,and obtained the three effective components of acute lung injury,acid,paeoniflorin and ethyl gallate.This paper laid the foundation for the further study of the pharmacological basis of Radix Paeoniae Rubra.