| BackgroundAlcoholic liver disease(ALD)is a relatively common liver disease in our country.The initial stage is alcoholic fatty liver disease(AFLD).AFLD is a relatively common liver disease,and it is the first step in the development of ALD to a more severe stage.At the same time,the incidence of AFLD is increasing every year.As a more common liver disease,AFLD is the first step in the development of ALD to a more serious stage,and the incidence of AFLD is increasing year by year,which seriously threatens people’s physical and mental health.Interleukin-8(IL-8),also known as CXCL8.IL-8is an effective inflammatory cytokine,which is involved in various experimental liver damage induced by LPS,TNF and bacterial infections.In ALD,the increase of IL-8 in plasma and liver has been well documented,and IL-8 is believed to play an important role in liver neutrophil infiltration.Previous studies have shown that IL-8 not only mediates a variety of inflammatory reactions,but also participates in a variety of liver diseases by binding to the CXCR1/2 receptor.The purpose of this experiment is to inject human IL-8(hIL-8)into alcohol liver mice through tail vein hypertension to observe the effect of Akt/HIF-1α pathway on related lipid gene metabolism Whether it affects and the degree of damage to the liver by oxidative stress.By understanding the biological activity of certain inflammatory factors,it may provide some new therapeutic ideas for the treatment of AFLD.MethodDue to the lack of IL-8 gene in mice,we used high-pressure injection to deliver10μg of pLIVE-hIL-8 plasmid to the liver of mice.The specific plasmid can be continuously expressed in the liver throughout the modeling period.Then the mice were divided into groups:(1)mice on a normal diet;(2)mice on a normal diet injected with hIL-8;(3)mice on a normal diet injected with an empty plasmid;(4)mice on an alcohol diet(5)Alcohol-fed mice injected with hIL-8;(6)Alcohol-fed mice injected with empty plasmid.Using the method of reference,we made a 16-day alcohol liver model in mice.In in vitro cell experiments,we added Akt inhibitor MK-2206 to inhibit the Akt /HIF-1αpathway.Then we use the kit to detect the liver-related biochemical indicators,western Blot and qRT-PCR and other methods to detect the key proteins and mRNA in the Akt /HIF-1α pathway and liver-related lipid genes.Then Western Blot was used to detect the expression of key proteins in the Akt/HIF-1α pathway and related lipid genes in liver cells.Results Compared with mice on a normal diet,the fatty vacuoles in liver tissue of mice fed alcohol diet increased,lipid accumulation increased,serum ALT,AST,TG,TCH biochemical indicators increased,and Akt/HIF-1α pathway protein The expression increased slightly,the expression of liver-related lipid gene sterol regulatory element binding protein-1c(SREBP-1c)increased,and the expression of peroxisome proliferator-activated receptor alpha(PPARα),which is related to liver fatty acid oxidation,decreased.Compared with the alcohol-fed mice injected with empty plasmids,the alcohol-fed mice injected with hIL-8 increased the number of fatty vacuoles in the liver tissues,and the accumulation of lipids was more serious,and the biochemical indicators of ALT,AST,TG,and TCH in the serum increased.The expression of Akt/HIF-1α pathway protein increased,the expression of SREBP-1C increased,and the expression of PPARα decreased significantly.The results of cell experiments found that when the Akt inhibitor MK-2206 was added to inhibit the Akt/HIF-1α pathway,the total protein did not change,but the expression of p-Akt and HIF-1α decreased,and the expression of SREBP-1c also decreased.On the contrary,the expression of PPARαdecreased.The expression has increased.At the same time,the expression of reactive oxygen species(ROS)in the liver tissues of mice injected with hIL-8 alcohol feed increased.Conclusion1.hIL-8 cytokine promotes the migration of neutrophils and increases infiltration in the liver of mice.2.hIL-8 cytokine increased the ROS level in the inflammation site and exacerbated the pathological degree of fatty liver disease in mice caused by alcohol.3.hIL-8 cytokines can aggravate the condition of alcohol-induced fatty liver mice through the Akt/HIF-1α pathway,and progress toward a more deteriorating stage. |
