Study On The Material Basis Of Poria Cocos Spleen Strengthening Effect Based On Spectrum Effect Relationship

Poria cocos is the dried sclerotia of the fungus Poria cocos(Schw)Wolf.Poria has the effects of strengthening the spleen,diluting water and dampening the heart,and has a wide range of clinical applications.Modern pharmacological studies show that Poria cocos has immunomodulatory,antioxidant,anti-tumor and other effects.Due to the lack of research on the effective substances and target of Poria cocos,especially the material basis and mechanism of its traditional effects are not clear,it is impossible to establish a complete quality of Poria cocos due to the lack of quality markers reflecting its effectiveness The evaluation system is also difficult to provide a strong scientific basis for the clinical use of traditional Chinese medicine.Therefore,under the guidance of the theory of Chinese medicine,this paper adopts the research idea of spectrum-effect characterization based on the integrity of traditional Chinese medicine to carry out the pharmacodynamics and identification of pharmacodynamic substances of the effect of Fuling and spleen.It aims to provide a scientific basis for the interpretation of the material basis and mechanism of Poria's traditional spleen-invigorating effect,and it also lays the foundation for the improvement of Poria's quality evaluation system.This thesis mainly completed the following research work.1.Taking 20 batches of Poria of different origins as the research object,the fingerprint of Poria cocos and the determination of the contents of 11 triterpenes were detected by UPLC method and dual-wavelength simultaneous detection.The fingerprint spectrum identified 32 common peaks,and the similarity analysis results showed that there were differences in 20 batches of samples from different origins;The total content of 11 compounds(16 ?-Hydroxydehydrotrametenolic acid,16?-Hydroxytrameteno lic acid,Poricoic acid B,Dehydrotumulosic acid,Poricoic acid A,Polyporenic acid C,3-O-Acetyl-16?-hydroxydehydrotrametenolic acid,Dehydropachymic acid,Pachymic Acid,Dehydrotrametenolic acid,Dehydroeburicoic acid)in 20 batches of samples ranges from 0.1078% to 0.1651%;The comprehensive scores obtained from the principal component analysis of the fingerprints of the 20 batches of samples are similar to the ranking of the sum of the contents of the 11 compounds.Based on the fingerprint information of 20 batches of samples,the content of 11 compounds,combined with the determination results of total triterpenes(0.24~0.78%)and water-soluble polysaccharides(0.51~2.59%),10 batches of samples with large differences were screened for the follow-up spectrum effect study of Poria cocos.2.The UPLC method was used to detect fingerprints of 10 batches of water extracts from Poria cocos(Hu Bei: Ying Shan(S1),Luo Tian(S2),En Shi(S3),Yun Nan: Chu Xiong(S4),Da Li(S5),Meng Hu(S6),He Na n:Shang Cheng(S7),Hunan: Huai Hua(S8),An Hui: Yue Xi(S9),Gui Zho u: De Jiang(S10))from different origins,and peak areas of 20 fingerpri nt peaks were obtained as the spectral efficiency analysis data.Using UP LC-Q-TOF-MS combined with reference substance comparison,17 chemi cal components in the common fingerprint peak were identified or inferr ed,which were 3-Oxo-16?hydroxylanosta-7,9(11),24-trien-21-oic acid,3-epidehydrotumulosic acid,3-Oxo-16?-acetyloxylanosta-7,9(11),24-trien-21-oi c acid,tumulosic acid,3-O-Acetyl-16?-hydroxytrametenolic acid,25?-Hydro xytumulosic acid and the above 11 compounds.3.3.A compound model of irregular diet plus overwork was used to establish a rat model of spleen deficiency.The spleen deficiency related indicators and HE staining results of stomach and small intestine tissues were used to evaluate the effect of water extracts of Poria cocos.The mechanism of Poria cocos spleen invigoration was preliminarily explored with relevant indexes such as brain-gut peptide,immune factor and aquaporin.The results of pharmacodynamic studies show that compared with the model group,S1 ~ S10 can promote the weight gain of rats and regulate the content of D-xylose,ACH,EGF,ALB and TP in rats with spleen deficiency.Among them,the levels of serum D-xylose and ACH in the S1 ~S10 groups were significantly increased(p < 0.01);Except for S2 and S4,the water extracts of Poria cocos were significantly increased the serum EGF content of rats(p < 0.01);Except S8,the water extracts of Poria cocos increased significantly the ALB content(p < 0.05);S1,S2,S4~S6 and S10 significantly increased the TP content(p < 0.05).The results of HE show that S1 ~ S10 have different degrees of protection on the gastrointestinal tissues of rats with spleen deficiency.The experimental results show that 10 batches of Poria cocos extracts from different places all have spleen-invigorating effects,but there are differences in the intensity of action.The research results of the effect of water extract of Poria on brain-gut peptide related indexes of spleen-deficiency rats showed that compared with the model group,the different water extract groups of Poria can increase the content of GAS,MTL and SP in the serum of spleen-deficiency rats to different degrees,Reduce the serum VIP and gastric tissue CCKAR,5-HT,CGRP content.The research results of the effect of water extract of Poria on immune factor related indexes of spleen deficiency rats showed that,compared with the model group,Poria all drug administration groups can reduce serum IFN-?,IL-1?,IL-6 in spleen deficiency rats,IL-17,TNF-? content and increased serum IL-2,IL-4,TGF-?1 content.In addition,compared with the model group,each administration group can reduce the levels of AQP1 and AQP2 in the renal medulla of spleen-deficiency rats to varying degrees,and they are very significantly different.From the above,Poria has a spleen-invigorating effect.However,there are differences in the efficacy of Poria cocos from different places,and there are also differences in the regulation of brain-gut peptides,immune factors and aquaporin-related indicators,and the subsequent correlation analysis of fingerprints and drug effects can be performed.4.AHP is used to determine the weight coefficient of each drug effect index in the total drug effect.The gray correlation method was used to analyze the correlation between each chemical component represented by each common peak in the fingerprint and each pharmacodynamic index,total pharmacodynamics,and indexes of brain-gut peptide,immune factor,and aquaporin.The results show that the correlation degree of the 20 chromatographic peaks with each drug efficacy index is greater than 0.6,indicating that there is a correlation,but there is a difference in the correlation degree of the same drug efficacy index with different chemical components;There is also a difference in the degree of correlation between the same chemical composition and different drug efficacy indicators.The correlation range between the 20 chromatographic peaks and the total drug efficacy ranges from 0.6964 to 0.8078,and The relevance of the top 3 is0.8012~0.8078(3#>14#(3-epidehydrotumulosic acid)>4#(25 ?-Hydroxytumulosic acid)),Showing that the correlation between the ingredients is not strong and Poria's role in strengthening the spleen is the result of the combination of its effective substance groups.The 17 components identified in the 20 chromatographic peaks are all triterpenoids,indicating that Poria triterpenes are effective components.The results showed that the correlation between the 20 chromatographic peaks and the 7 brain-gut peptide indexes were all greater than 0.65,indicating that they were all correlated.The correlation analysis results with 8 immune factor indicators show that 20 chromatographic peaks are correlated with IFN-?,IL-1?,IL-17,TGF-?1,IL-2,IL-4,TNF-?,but only There are 15 peaks associated with IL-6,and the correlation is low.According to the data comparison,the triterpene chemical components in the 20 chromatographic peaks are highly correlated with the brain-gut peptide index,and the number of peaks is more than the immune factors.The conclusion is consistent with the conclusion that the triterpene component of Poria cocos mainly plays the role of spleen strengthening by regulating the level of brain-gut peptide factor,which It is the conclusion of another experiment carried out by the research group called ‘The Pharmacodynamics and Mechanism of Different Extraction of Poria cocos on tonifying spleen'.To sum up,the research in this paper interprets the material basis of Poria cocos to strengthen the spleen,and the conclusion lays the foundation for determining the quality of Poria cocos,and also provides a scientific basis for the clinical application of Chinese medicine.