| Objective:In this study,to evaluate the effects of n-3 polyunsaturated fatty acid(n-3 PUFA)on oxidative stress,inflammation and lipid metabolism in obese mice induced by high-fat diet,Fad3 transgenic mice was used.Transgenic FAD3 mice have been genetically engineered to ubiquitously express the Fad3 n-3 fatty acid desaturase from Arabidopsis thaliana.Methods:1.C57BL/6 mice and Fad3 transgenic mice were divided into four groups C57BL/6C group(n=10),C57BL/6H group(n=10),FAD3C group(n=10)and FAD3H group(n=10).C57BL/6C group and FAD3C group were fed 10%kcal diet,C57BL/6H group and FAD3H group were fed 45%kcal high fat diet,for 40 weeks in total.2.After the mice were sacrificed,the liver tissues were collected,MDA,SOD,GSH,GSSG,GSH/GSSG,NOX2 were measured,and the oxidative damage and antioxidant status were evaluated.3 The expression level of inflammatory signaling pathway related proteins(NF-?B p65 and p-NF-?B p65)were detected by western blotting,and the mRNA expression level of IL-6 and TNF? were measured by real-time quantitative PCR.4.The protein expression of AMPK,p-AMPK,SREBP1-c and p-ACC were detected by western blotting,and the mRNA expression of PPAR?,CPT-1,ACOX and FAS were measured by real-time quantitative PCR Results:1.The body weight and body fat of C57BL/6H group were significantly higher than those of the other three groups(P<0.01).There was no difference in food intake among the four groups.2.Parameters related to liver oxidative stress were detected.The MDA content of C57BL6H group was significantly higher than that of the other three groups(P<0.05),while that of FAD3H group was significantly lower than that of the other three groups(P<0.05)SOD activity of C57BL/6H group was significantly lower than that of other three groups(P<0.05),and there was no significant difference among C57BL/6C,FAD3C and FAD3H groups GSH level of C57BL/6H group was significantly lower than that of C57BL/6C and FAD3H group(P<0.01),but there was no significant difference between FAD3H group and C57BL/6C group.GSSG level of four groups was not significantly different.GSH/GSSG level in C57BL6H group was significantly lower than that in other three groups(P<0.05)There was no significant difference in the expression of NOX2 protein between C57BL6C group and C57BL6H group.The expression of NOX2 protein in FAD3C group and FAD3H group decreased significantly(P<0.01).3.Expression of genes,related to inflammatory signaling pathway,were tested in liver.There was no significant difference in p65 protein expression among the four groups.P-p65 protein expression in C57BL6H group was significantly higher than that in other three groups(P<0.05).The expression level of p-p65/p65 expression in FAD3H group was significantly lower than that in C57BL6H group(P<0.05).TNF? mRNA expression in C57BL6H group was significantly higher than that in other three groups(P<0.05).IL-6 mRNA expression in FAD3H group was significantly higher than that in other three groups(P<0.05).4.Expression of genes which related lipid metabolism were detected in liver.The expression level of p-AMPK protein in C57BL6H group was significantly lower than that in C57BL6C group(P<0.05).And level of p-AMPK protein in FAD3H group was significantly higher than that in C57BL6H group(P<0.05)There was no significant difference in the expression of AMPK protein in the four groups The trend of p-AMPK/AMPK was the same as that of p-AMPK.The expression of PPAR?mRNA in C57BL6H group was significantly lowest than that in other three groups(P<0.01)PPAR? mRNA expression in FAD3C group and FAD3H group increased significantly(P<0.05).The expression of CPT-1 mRNA in FAD3C group was significantly higher than that in other three groups(P<0.01).The expression of ACOX mRNA in C57BL6H group was significantly higher than that in other three groups(P<0.05).The expression of SREBP1-c protein and FAS mRNA in C57BL6H group were significantly higher than those in other three groups,while that in FAD3H group was significantly lower than that in C57BL6H group(P<0.05).The expression level of p-ACC protein in C57BL6H group was significantly lowest in four groups(P<0.05).And p-ACC protein expression of FAD3H group was significantly higher than that in C57BL6H group(P<0.05).Conclusion:1.n-3 PUFA reduced the body weight and body fat weight of obese mice induced by high fat food.2.n-3 PUFA prevented the increase of ROS mediated by NADPH oxidase by down regulating the expression of NOX,and reduced the content of MDA to reduce the oxidative stress of liver Up-regulated the antioxidant system,such as increasing the expression of SOD,GSH,GSH/GSSG,to improve the antioxidant capacity of liver.3.n-3 PUFA attenuated inflammatory stimulation by down regulating inflammatory signaling pathway related genes,such as p65,IL-6,TNF?.4.By activating AMPK,n-3 PUFA down-regulated the expression of SREBP1-c and FAS,up-regulated the expression of p-ACC,PPAR?,ACOX and CPT-1,and improve liver lipid metabolism. |
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