Study On The Extraction Technology Of Lonicera Edulis Anthocyanin And The Treatment Mechanism Of Xerophthalmia

Lonicera edulis is a wild edible berry with high nutritional value,which has broad development prospects.In this paper,the anthocyanins were extracted from the fruit of Lonicera edulis by ultrasonic wave,and then purified with macroporous adsorption resin.HPLC-MS/MS was used to identify the anthocyanin of compounds.The anti-inflammatory activity of Lonicera edulis anthocyanins was analyzed based on molecular docking.And the mechanism of action and activityof Lonicera edulis compound tablet in the treatment of xerophthalmia was analyzed based on network pharmacology.The research contents were showed as following:The optimal process for extracting Lonicera edulis anthocyanins through single factor experiment and response surface method is as follows:ultrasonic power 160 W,ultrasonic time 55 min,material liquid ratio at 1:35 g/m L,and the ethanol volume fraction 85%.The extraction of anthocyanins form Lonicera edulis was（298.22±1.13）mg/100g.The static Lonicera edulis anthocyanins adsorption-desorption characteristics in 4 kinds of macroporous resinswere compared,and the dynamic Lonicera edulis anthocyaninsadsorption-desorption conditions in AB-8 type macroporous resins were studied,and the purification conditions were determined as follows:sample solution-p H 3.0,sample flow rate-1.0 m L/min,the eluent ethanol concentration-80%,and the eluent flow rate-1.0 m L/min.Under these conditions,the content of lonicera edulis anthocyanins is（317.35?0.82）mg/g.Four kinds of Lonicera edulis anthocyanins were identified by HPLC-MS/MS,which were Cyanidine-3-hexoside-catechin,Cyanidine-3-hexoside dimer,Cyanidine-3,5-dihexoside and Cyanidine-3-glucoside,respectively.Among them,the content of cyanidine-3-glucoside is the highest（68.971%）.The anti-inflammatory mechanism of Lonicera edulis anthocyanins was analyzed based on molecular docking.Construct the"component-target-pathway"network and target interaction network with xerophthalmia through Lonicera edulis compound tablet（Lonicera edulis,chrysanthemum,medlar）Analyze its mechanism of action in the treatment of xerophthalmia.The results showed that the 4 active ingredients of Lonicera edulis and the 7 active ingredients screened by oral bioavailability and drug-like properties of chrysanthemum and wolfberry,they regulates 10 key targets and 10 main signal pathways for the treatment of xerophthalmia,forming a multi-ingredient;multi-target,and multi-channel Network characteristics with integrated regulation effect.The mechanism of Lonicera edulis compound tablet in the treatment of xerophthalmia is to inhibit the occurrence and development of xerophthalmia by regulating the expression of potential targets and regulating pathways related to inflammation and cell apoptosis.LPS was used to stimulate RAW264.7 to create a cell inflammation model.Use MTT method to detect cell survival rate,investigate the release amountof NO,PGE₂,IL-6 and IL-1?in RAW264.7 at different doses of Lonicera edulis compound tablet.The results showed that when the doses of drug were 50μg/m L,100μg/m L,200μg/m L,and 400μg/m L,the cell survival rate was maintained above 90%,indicating that it has no toxic effect on cells.Griess method was used to detect the release amount of NO in RAW264.7 cells induced by LPS,and ELISA method was used to detectthe release amount of IL-1?,IL-6 and PGE₂in RAW264.7 cells induced by LPS.Each dose of drug has a significant inhibitory effect on the release levels of NO,PGE₂,IL-6 and IL-1?in the model group（P<0.05）,indicating that Lonicera edulis compound tablet can reduce inflammation by reducing inflammation to relieve the occurrence and development of xerophthalmia.