Quality Evaluation Of Eucommia Cortex And Study On Anti-Osteoporosis Chemical Constituents Of Salt Eucommia Cortex

Eucommiae Cortex is the dry bark of Eucommia ulmoides Oliv,a single family and single genus plant.It has the effects of tonifying liver and kidney,strengthening muscles and bones,and preventing pregnancy.As early as 2000 years ago,people had a deep understanding of the efficacy and application of Eucommiae Cortex.It is widely used in clinical practice,and is often used in liver and kidney tonic prescriptions or alone.Salt Eucommiae Cortex is a commonly used decoction of Eucommiae Cortex.After salt moxibustion,it can enhance its effect of tonifying liver and kidney and strengthening muscles and bones.It is the main processed product of Eucommia Eucommiae Cortex.In this paper,firstly,the quality of Eucommiae Cortex was studied,and the quality grade standard of Eucommiae Cortex,was established based on the debate of quality,and the processing of Eucommiae Cortex was studied.The mechanism of enhancing anti-osteoporosis after processing was explored by network pharmacology and cell experiment.The research contents are as follows:(1)Study on the quality of Eucommiae Cortex: collect 28 batches of Eucommiae Cortex that meet the requirements of the pharmacopoeia,establish a TLC identification method based on the existing quality standards,establish a HPLC method for the determination of six components of aucubin,geniposide,chlorogenic acid,geniposide,geniposide and pinoresinol diglucoside,and establish a fingerprint of Eucommia ulmoides,with 10 common peaks,Five of the common peaks were identified with the reference substance.To form the quality improvement standard of Eucommia ulmoides and better control the quality of Eucommia ulmoides.(2)Research on the quality grade standard of Eucommia Cortex medicinal materials: On the basis of the existing specifications and grades of Eucommia Cortex,according to the traditional perspective of identifying high-quality Eucommia Cortex,the external indicators such as eucommia gum content,internal and external epidermal color,thickness and other external indicators were determined,the intrinsic component indexes of geniposidic acid,genipin,geniposide and pinoresinol diglucoside were determined,and the Pearson correlation analysis of traits and intrinsic content was carried out by using multiple statistical analysis techniques,and the indicators were screened by combining principal component analysis,orthogonal partial least squares discriminant analysis and K-mean cluster analysis Combined with the feasibility of market application,the medicinal materials of Eucommia are divided into three grades.(3)Research on the changes of components before and after Eucommia Cortex salt moxibustion: According to the general principles of the 2020 edition of the Chinese Pharmacopoeia,the study of Eucommia Cortex chinensis was carried out.A comprehensive analysis of the components before and after Eucommia Cortex salt moxibustion was carried out by LC-MS technology,and a total of 48 components were analyzed,and according to the LC-MS ion peak area comparison,the content of12 components increased significantly,most of which were components that were thermally decomposed glycosides,such as turpentinol,medium lipids,etc.,which produced two new components,5-HMF and ferulaldehyde.In the process of processing,samples were taken at different time points,and the contents of water and extract,pinoresinol diglucoside,chlorogenic acid,genipin,geniposide and geniposidic acid at different times were monitored,and it was found that the content of the extract of the three batches of medicinal materials increased by an average of 2.71%,and the rest decreased significantly to 25 min,and the average decrease of geniposidic acid,geniposide,genipin,pinoresinol diglucoside and chlorogenic acid decreased by43.80%,59.49%,72.09%,38.31% and 16.38%。(4)Network pharmacology research and in vitro verification experiment on the preparation of enhanced anti-osteoporosis by eucommia chinensis: Based on the results of chemical composition changes of Eucommia Cortex before and after salt moxibustion,for the components whose content significantly increased(>5 %)after processing,the five principles of Lipinski drugs were first screened to obtain chemical components with high oral availability,including 5-hydroxymethylfurfural,coniferaldehyde,caffeic acid,cycloolivil,pinoresinol,olivil and syringaresinol.Pharm Mapper was used to screen potential human targets,and Gene-Cards database and OMIM were used to query OP-related targets,and the two types of targets were intersected to obtain potential targets that enhanced the anti-osteoporosis effect after eucommia was concocted.The data were imported into the STRING online database to analyze the protein interaction between the targets,the results were imported into Cytoscape 3.9.0 in csv.format and topological analysis,and the key active components coniferaldehyde and pinoresinol were screened according to the degree value,and finally the mouse embryonic osteoblast(MC3T3-E1)was used to verify that ferulic aldehyde and turpentol can promote the proliferation and differentiation of osteoblasts,and achieve the effect of reinforcing liver and kidney,strengthening muscles and bones.To sum up,this study has completed the draft of the quality standard improvement of Eucommia Cortex and formed the quality grade standard of Eucommia Cortex,so as to better control the quality of Eucommia Cortex,and provide reference for the high quality and good price of Eucommia Cortex in the market;The content of most components of Eucommia Cortex after processing decreased due to thermal decomposition,and many glycosides were obtained.According to the network pharmacological screening prediction,the seven components of 5-HMF,coniferaldehyde,caffeic acid,cycloolivil,pinoresinol,olivil and syringaresinol were the important components of Eucommia Cortex after processing to play an anti-osteoporosis role,and were verified by osteoblasts(MC3T3-E1),It was found that ferulic aldehyde and oleoresin could promote the proliferation and differentiation of osteoblasts.It shows that Eucommia salt moxibustion can increase the content of ferulic aldehyde and oleoresin,promote the proliferation and differentiation of osteoblasts,and achieve the effect of reinforcing liver and kidney,strengthening muscles and bones.