Analyses Of Fatty Acid Components Of Flaxseed Oil And Its Effects And Mechanisms On Alleviation Of Alcoholic Liver Disease In Mice

Linum usitatissimum L.isthe genus Linum in the Linaceae family.Flaxseed is one of top 10oil-baering crop seeds.Flaxseed oil represents a edible oilwith high quality,which is rich in unsaturated fatty acids(UFAs),especially ?-3 ?-linolenic acid.Accelerating studies have demonstrated the beneficial effects of flaxseed oil for health in many aspects.However,the difference of inflaxseed oil from different habitatsamong oil contents,components of fatty acids and mineral contents still remains unclear.In the present study,these indicators in flaxseed oil from different habitats were detected and compared,respectively.Furthermore,the effects of flaxseed oil on alcoholic liver disease wereinvestigated in mice.The main results were as follows:(1)In the present study,oil contents and components of fatty acids inflaxseed oil from 6 different habitats were measured and ananlyzed using Soxhlet's extraction and GS-MS,respectively.Our results showed that oil contents of flaxseeds from different habitats ranged from 36.59 to 44.88%.Further analyses revealed that oil content was negatively correlated with accumulated growing season temperature(r=-0.840,P=0.037).Fatty acids of flaxseed oils were mainly composed of linolenic acid(53.36-65.84%),linoleic acid(10.14-16.39%),oleic acid(10.03-12.37%),stearic acid(3.98-9.85%)and palmitic acid(2.41-7.97%).The contents of UFAsin flaxseed oil were up to 77.51-92.39%.According to oil contents and components of UFAs,flaxseed oil from Longnan District exhibited significantadvantages.Thestudy may contribute to the exploitationof edible and medical value of flaxseed oil.(2)Mineral contents of flaxseed oil from 6 different habitats were analyzed by ICP-AES.The results showed that Fe,Na,Ca,P,and Mg as principal component in flaxseed oil,showed significant difference among different habitats,demonstrating that flaxseed oil represents a edible oil with high qualityf.(3)Alcoholic liver disease(ALD)isa chronic wide-spectrum of liver injury caused by consistently excessive alcohol intake.Few satisfactory advances have been made in themanagement of ALD.Thus,novel and more practical treatment options are urgently needed.Flaxseed oil(FO)is rich in a-linolenic acid(ALA),plant-derived n-3 polyunsaturated tatty acids(PUFAs).However,the impact of dietary FO on chronic alcohol consumption remains unknown.In this study,we assessed possible effects of dietary FO on attenuation of ALD and associated mechanisms in mice.Firstly,mice were randomly allocated into four groups:pair-fed(PF)with corn oil(CO)group(PF/CO);alcohol-fed(AF)with CO group(AF/CO);PF with FO group(PF/FO);AF with FO group(AF/FO).Each group was fed modified Lieber-DeCarli liquid diets containing isocaloric maltose dextrin as a control or alcohol with corn oil and flaxseed oil,respectively.After 6 weeks of feeding,mice were euthanized and associated indicatorswere investigated.Our result showed that body weight(BW)in AF/FO group was significantly elevated,compared with AF/CO group.Dietary FO reduced the abnormal elevated levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in chronic ethanol consumption.Amelioration of these parameters as well as liver injury via HE staining in dietary FO supplementation in ALD demonstrated that dietary FO can effectively benefit for the protection against ALD.To further understand the underlying mechanisms,we investigated the inflammatory cytokines and gut microbiota.A series of inflammatory cytokines,including TNF-?,IL-1?,IL-6 and IL-10,were determined.As a result,TNF-?,IL-1? and IL-6 were significantly decreased in AF/FO group compared with control group(P<0.05);IL-10 showed no significant alteration between AF/CO and AF/FO groups(P>0.05).The results of western blot,immunohistochemistry and flow cytometry demonstrated that flaxseed oil reduced the hepatic kupffer cells,especially Ml subset.Taken together,flaxseed oil can ameliorate alcoholic liver disease via anti-inflammation.(4)Sequencing and analysis of gut microbiota gene indicated that a reduction of Verrucomicrobia,Actinobacteria,Porphyromonadaceae unclassified,Parasutterella,Akkermansia,as well as an increase in Firmicutes,Deferribacteres,Parabacteroides,Ruminococcaceae unclassified,Bacteroides,Lachnospiraceae_unclassified,Alistipes,were seen in AF group compared with PF control.Furthermore,dietary FO in ethanol consumption group induced a significant reduction in the phylum level included Proteobacteria and Deferribacteres,compared with AF/CO group.In the genus level,dietary FO altered Bacteroides,Parabacteroides as well as Alistipes.Collectively,dietary FO ameliorates alcoholic liver disease via anti-inflammation and modulatinggut microbiota,thus can potentially serve as an inexpensive intervention for the prevention and treatment of ALD.