Study On The Protective Effect Of Syringin On D-GalN/LPS Induced Liver Injury In Mice

Syringin has pharmacological effects such as anti-inflammatory,antioxidant,and it has great potential value.In this paper,D-(+)-galactosamine combined with lipopolysaccharide was used to establish an acute liver injury model,and various methods such as network pharmacology,molecular docking,q RT-PCR,immunohistochemistry,pathological tissue analysis,and intestinal flora analysis were used to evaluate the The hepatoprotective effect of syringin and its mechanism of action.The specific methods are as follows:1: Analysis of potential liver-protective targets of syringin by network pharmacology.A total of 2251 disease targets with "Drug-induced liver injury" as keywords and 116 drug targets with "syringin" as keywords were obtained after correcting and deweighting using databases Uniprot,Genecards,and OMIM.Sixty-four drug-disease common targets were obtained after the intersection in Venn software.Based on the Degree value,the core targets in the conjunction were ranked,and AKT1 was identified as the target protein with the highest Degree.The molecular docking results showed that the syringin could bind tightly to AKT1 with a docking score of-7.55 kcal/mol.2: To establish a D-Gal N/LPS liver injury model and initially explore the hepatoprotective effect of syringin.Forty experimental mice were divided into four groups,normal group(NC group),model group(LD group),syringin low dose group(LSY+LD group),and syringin high dose group(HSY+LD group).Three days before the constructing model,the LSY+LD group and HSY+LD group were continuously injected with the corresponding doses of syringin at 25 mg/kg and 50 mg/kg,respectively.1h after the administration on the third day,the normal group continued to be injected intraperitoneally with saline.In comparison,the remaining three groups were injected intraperitoneally with D-Gal N/LPS(250mg/kg+30 μg/kg)to construct a liver injury model.6 h after the attack,the required mice serum,as well as liver and intestine tissue samples,were taken.The levels of ALT and AST in mice serum,the levels of SOD,GSH,and CAT in liver tissues,and the levels of inflammatory factors IL-1β,IL-6,and TNF-α were measured using the kit.The results showed that compared with the NC group,the serum levels of ALT and AST were significantly higher(P < 0.01),the levels of inflammatory factors IL-1β,IL-6,and TNF-α were substantially higher(P < 0.01),and the stories of SOD,GSH,and CAT were significantly lower(P < 0.01)in the liver tissues of mice in the LD group,compared with the LD group,the levels of ALT and AST in the LSY+LD group were significantly higher(P <0.01),and the stories of inflammatory factors IL-1β,IL-6,and TNF-α in the HSY+ LD group,ALT and AST were significantly decreased(P <0.01),IL-1β,IL-6,and TNF-α levels were decreased(P < 0.05,P < 0.01),and SOD,GSH,and CAT levels were increased to different degrees(P <0.05,P < 0.01).The results also showed that syringin could alleviate the pathological changes of liver tissue and inhibit NF-κBp65 activation and nuclear translocation in LD group mice.This section demonstrates the successful establishment of the liver injury model and the preliminary evidence that syringin can attenuate the pathological condition of DGal N/LPS-induced liver injury.3: Investigating the anti-apoptotic effect of syringin.Comprehensive analysis of AKT1 upstream and downstream anti-inflammatory,antiapoptotic,and other related signaling pathways,and detection of caspase-3,caspase-8,caspase-9,BCL-2,BAX,and other proteins in mouse liver using ELISA kits,q RT-PCR detection of BCL-2,BAX,Caspase-3,TNF-α,IL-1β,IL-6 m RNA levels in mice liver.Immunohistochemical was performed to analyze the nuclear translocation rate of p-Nrf-2,cleaved caspase-3,and TUNEL to analyze the strength of apoptotic signals in liver tissues.The results showed that the levels of caspase-3,caspase-8,caspase-9,and BAX proteins were significantly increased(P < 0.01).Their corresponding m RNA levels were significantly increased(P < 0.01,P <0.05)in the tissues of mice in the LD group,and the stories of BCL-2protein were significantly decreased(P < 0.01).Its m RNA level was also considerably reduced,and syringin was able to reverse the above changes in protein levels and m RNA levels(P < 0.01,P < 0.05).The syringin group was able to reduce apoptotic signals,inhibit nuclear translocation of cleaved caspase-3,and promote nuclear translocation of p-Nrf-2 in liver tissues of the LD group.The above results demonstrate that syringin can achieve hepatoprotective effects through anti-inflammatory,antioxidant,and anti-apoptotic pathways.4: Based on the previous sections,we further investigated the effect of syringin on the flora of mice’s colon intestinal contents.16 Sr DNA sequencing showed that D-Gal N/LPS could increase the abundance of harmful bacteria Lachnospiraceae in mouse colon(P < 0.05),syringin could increase the number of beneficial bacteria in the colon intestine,such as Firmicutes,Lactobacillaceae(P < 0.05),Lactobacillus and The abundance of Alistipes in the colon intestine was increased,and the lot of Lachnospiraceaewas decreased.In conclusion,this study demonstrated that lilac glycosides could protect against liver injury through anti-D-Gal N/LPS-induced inflammation,oxidative stress,and apoptosis in response to liver damage.In addition,lilac glycosides can promote the proliferation of beneficial bacteria in the colonic intestine and reduce the abundance of harmful bacteria,etc,to further reduce D-Gal N/LPS-induced liver injury through the intestine-liver axis.