Screening Of Liver-Protecting Active Parts Of Ligusticum Chuanxiong And Preliminary Study On Its Preparation

Chuanxiong has the effects of promoting blood circulation,removing blood stasis,dispelling wind and relieving pain.In recent years,studies have found that certain monomer components with hepatoprotective effects are also present in Ligusticum chuanxiong medicinal materials,but the screening of active parts of Ligusticum chuanxiong hepatoprotective and the development of related preparations have not been studied.This article intends to use a combination of pharmacology and chemistry to study the material basis of the liver-protecting active parts of Ligusticum chuanxiong.At the same time,the liver-protecting preparations of Ligusticum chuanxiong are prepared and preliminary evaluated.It is expected to lay the foundation for the basic research of the liver-protecting medicinal material of Ligusticum chuanxiong and the development of related new products.The research methods and results mainly include the following four aspects.1.Screening of active parts of Ligusticum chuanxiong for protecting liverThe in vivo pharmacodynamic method was used to screen the sites with the best liver-protecting activity of Ligusticum chuanxiong.First,extract the volatile oil of Ligusticum chuanxiong,Ligusticum chuanxiong polysaccharide and ethanol extract of Ligusticum chuanxiong extract from the decoction pieces of Ligusticum chuanxiong.Establish a mouse model of acute liver injury,measure the levels of alanine aminotransferase and aspartate aminotransferase in serum to evaluate the degree of liver damage;measure malondialdehyde and superoxide dismutase in liver tissues to evaluate the antioxidant activity of various parts in vivo;preparation Pathological section of the liver to observe the damage and repair of the liver.By comparing the efficacy results of different extraction parts,it was preliminarily screened out that the ethanol extraction part of Ligusticum chuanxiong had better hepatoprotective effect.The resin separation technology is used to further separate the water elution part and the ethanol elution part from the ethanol extraction part of Ligusticum chuanxiong.A mouse model of acute liver injury was also established to compare the pharmacodynamics of different elution sites,and it was determined that the site where Ligusticum chuanxiong exerted its hepatoprotective efficacy was mainly the ethanol elution site.This research laid the foundation for the research on the material basis of Ligusticum Chuanxiong to protect the liver.2.Research on the material basis of the liver-protecting active parts of Ligusticum chuanxiongThe liquid-mass spectrometry analysis method and network pharmacology were used to study the material basis of the liver-protective active parts of Ligusticum chuanxiong.The liquid-mass spectrometry method analyzed that there are 39 chemical components in the liver-protecting active parts of Ligusticum chuanxiong,among which phenolic acids are the main components.The network pharmacological component target database was used to screen out 27 active components and their targets of Ligusticum chuanxiong hepatoprotective.The component targets are compared with the liver injury disease targets obtained from the screening of the disease gene database to obtain the intersection targets,and then they are imported into the STRING database for PPI analysis to obtain the core targets.GO enrichment analysis and KEGG analysis were performed on the core target.Finally,the selected active chemical components,targets and pathways were imported into Cytoscape 3.7.2software,and a network diagram of the ”component-target-pathway” of the ethanol elution part of Ligusticum chuanxiong was constructed.The research in this chapter has screened out 14 hepatoprotective active ingredients,19 core targets,and 10 pathways in Ligusticum chuanxiong that are directly or indirectly related to the treatment of liver injury.This study predicts the potential pathways of Ligusticum chuanxiong to treat liver injury,and provides a basis for further research on the mechanism.3.Study on the preparation of the active part of Ligusticum chuanxiong to protect the liverUsing ferulic acid as a reference substance,a UV analysis method of total phenolic acid in ethanol eluted part was established.The yield and encapsulation rate are used as evaluation indicators to prepare clathrates to improve the shortcomings of the ethanol elution site.Then,by investigating the preparation process of the tablet,the preparation process of the active part of Ligusticum chuanxiong hepatoprotective was determined.The linear relationship of the UV analysis method established in this study is good in the range of 0.0663～0.2652 mg/mL,and the content of total phenolic acid in the ethanol elution part of Ligusticum chuanxiong is 28.36%.The optimal process for preparing the hepatoprotective active parts of Ligusticum chuanxiong into the clathrate is that the feed ratio is 10:1,the rotation speed is 300 r/min,the stirring temperature is 60 ℃,and the stirring time is 2 h.The quality of the inclusion compound of Ligusticum chuanxiong prepared under the optimal process was evaluated: the clathrate was a light yellow,fine and uniform powder,and the elution position of Ligusticum chuanxiong alcohol could be seen from optical micrographs,differential scanning calorimetry analysis and infrared spectroscopy.It is well enclosed.Finally,it was determined that the optimal preparation process of the tablet is 61% as the main drug clathrate,38% as the filler lactose,and the powder is mixed and granulated under a 20-mesh sieve by wet granulation.After drying at low temperature,1% magnesium stearate is added.Press the tablet after uniform mixing.In this study,tablets with a smooth appearance and uniform color were prepared from the difficult-to-form bulk drug through the preparation of intermediates,which improved the shortcomings of the bulk drug itself,and also provided a reference for the molding of traditional Chinese medicine extract preparations.4.Study on the quality standard of Chuanxiong Liver-Protection TabletsTaking the relevant provisions of the 2020 edition of the Chinese Pharmacopoeia as a reference,the prescription and preparation method of Chuanxiong Liver-Protection Tablets are stipulated,and corresponding tablet inspections are carried out in accordance with the regulations.At the same time,the thin-layer identification method and content determination method of Chuanxiong Baogan Tablets were established,and the content of Liver-Protection Tablets was regulated.Ligusticum chuanxiong was extracted and purified by adding hydroxypropyl-β-cyclodextrin water saturated solution to prepare the clathrate,which was prepared according to the optimal prescription of Liver-Protection Tablets.Chuanxiong Liver-Protection Tablets have a complete and smooth appearance,uniform color,and gray-yellow color.When performing tablet inspections(differences in tablet weight,friability,and disintegration time limit)on Liver-Protection Tablets,the inspection items are in compliance with the pharmacopoeia.The development system of the thin-layer identification method is:cyclohexane-dichloromethane-ethyl acetate-formic acid(4:1:1:0.1),under ultraviolet light(365nm),the sample should be combined with ferulic acid.The reference substance showed spots of the same color at the same position.After establishing the content determination method,the determined content of ferulic acid in this product shall be the upper limit of the floating limit.For the liver protection tablets with a tablet weight of 0.50 g,the content of each tablet should not be less than 0.6 mg in terms of ferulic acid.