Study On The Preparation Of Acanthopanax Senticosus Glycosides Microemulsion And Its Anti-fatigue Mechanism

Objective: Acanthopanax senticosus glycosides are used as the anti-fatigue components of Acanthopanax senticosus.The mechanism of action is not clear,and the low bioavailability of Acanthopanax senticosus glycosides limits its development and utilization.This article analyzes the chemical substance basis for the anti-fatigue effect of Acanthopanax senticosus glycosides,and develops a microemulsion preparation based on the poorly fat-soluble physicochemical properties of the ingredients to enhance the permeability of the intestines and improve the biological properties of Acanthopanax senticosus glycosides utilization.The anti-fatigue mechanism of Acanthopanax senticosus glycosides was predicted by means of network pharmacology,which provided a scientific basis for Acanthopanax senticosus "long-term service,light body and old age".Methods: Comparing the antioxidant activity of Acanthopanax senticosus glycosides and the positive control drug vitamin C chewable tablets by the drug’s ability to scavenge DPPH free radicals and total reduction capacity;Analyze the chemical components of Acanthopanax senticosus glycosides based on liquid-mass spectrometry technology,at the same time,high performance liquid chromatography is used to establish a quality control method for the Acanthopanax senticosus glycosides;Use the D-optimal mixture design method to optimize the formulation and preparation process of Acanthopanax senticosus glycosides microemulsion based on the single factor method;Acanthopanax senticosus glycosides microemulsion for quality evaluation;In vivo one-way intestinal perfusion model to study the intestinal absorption characteristics of Acanthopanax senticosus glycosides and Acanthopanax senticosus glycosides microemulsion;Predict the Acanthopanax senticosus glycosides based on network pharmacology methods and molecular docking technology key targets and related pathways for anti-fatigue effects.Results: Acanthopanax senticosus glycosides have a stronger ability to scavenge DPPH free radicals than vitamin C chewable tablets,and the total reduction ability is equivalent to that of vitamin C chewable tablets;Based on the liquid-mass spectrometry technology,30 compounds in the Acanthopanax senticosus glycosides were identified,mainly including glycosides and a small number of organic acids;With eleutheroside B and eleutheroside E as the index components,the quality control method of Acanthopanax senticosus glycosides was established,and the content of eleutherococcus B must not be less than 0.90%,and the content of eleutherococcus E must not be less than 3.60%;Determine the preparation process of Acanthopanax senticosus glycosides microemulsion: the mixed surfactant is a mixture of absolute ethanol and soybean phospholipid(mass ratio 1:1),the mass percentage is 0.480,the oil phase is isopropyl palmitate,and its mass percentage is 0.312,and the water phase is an aqueous solution of Acanthopanax senticosus glycosides with a concentration of 20mg/m L,and the mass percentage is 0.208;The appearance of the prepared Acanthopanax senticosus glycosides microemulsion is yellow transparent and homogeneous liquid,standing without stratification.Observed by transmission electron microscope,its shape is uniformly dispersed,non-adhesive,uniform and rounded spherical emulsion;The type of microemulsion is water-in-oil type,at room temperature,the p H value is 5.19.the refractive index is 1.4165,the average particle size is 26.47 nm,and the PDI value is 0.118;The average content of eleutheroside B is 0.0389mg/m L,and the eleutheroside E 0.1664 mg/m L;The microemulsion is clear and transparent,no layered emulsion within 30 days,the content is basically unchanged,and the stability is good;The results of in vivo unidirectional intestinal perfusion show that the eleutheroside B,eleutheroside E in the Acanthopanax senticosus glycosides and microemulsion is absorbed in the duodenum,jejunum,ileum,and colon.The absorption of each intestinal segment is improved after the microemulsion is made,especially in the ileum.It is the best absorption site for microemulsion.It is speculated that the microemulsion is ingested by the abundant lymph nodes in the ileum and transported via the lymphatic pathway;Network pharmacological prediction results show that the core components of eleutheroside B,eleutheroside B1 and eleutheroside C are the core components that play an anti-fatigue effect in the Acanthopanax senticosus glycosides,the key targets are HSP90AA1,VEGFA,STAT3,MMP9.The molecular docking results show that the binding ability of the core components and key targets is similar to that of the positive drug Adenosine Disodium Triphosphate.Biological processes such as reaction and hypoxia response play an anti-fatigue effect by regulating HIF-1,PI3K-Akt,insulin signaling pathway and material metabolism.Conclusion: Based on the analysis of the chemical components of Acanthopanax senticosus glycosides,the material components of Acanthopanax senticosus glycosides were explored.The preparation and quality evaluation research of Acanthopanax senticosus glycosides microemulsion lays the foundation for the innovative development of new preparations of Acanthopanax senticosus medicinal materials.The prediction of potential targets and related pathways of the anti-fatigue effect of Acanthopanax senticosus glycosides provides scientific basis and reference for the research on the anti-fatigue mechanism of Acanthopanax senticosus.