Effects Of N-3PUFAs On Autophagy And Inflammation Of Hypothalamus And Body Weight In Mice

OBJECTIVE: Fat-1 transgenic mice were used as models to investigate the effects of endogenous n-3PUFAs on body weight,hypothalamic inflammatory factors and autophagy protein expression in mice,and to explore its mechanism.METHODS: The 6 week old mice were divided into two groups: fat-1 transgenic mice and wild type mice.The body weight and body length of 14 week old mice were measured,and the Lee's index was calculated.The frozen section of mice liver was stained with oil red O,and the lipid droplets in the liver of the two groups were observed.Electron microscopy was used to observe the autophagosome in the hypothalamic neurons of the two groups of mice.The autophagy proteins P62,LC3 and ATG7 were located in hypothalamus by immunofluorescence technique.The expression of P62,LC3 and ATG7 in the hypothalamus were quantitatively analyzed by western blot.The m RNA expression of TNF-?,IL-6,IL-1?,NF-k B,chemokine MCP-1,CCL5,CXCL12,CX3CL1,microglia markers TMEM119,GFAP were detected by real-time fluorescence quantitative PCR for relative quantitative analysis.Meanwhile,quantitative analysis of inflammatory factors TNF-alpha,IL-6,IL-1 beta,IFN-gamma,TLR-4,chemokine MCP-1,and m RNA of adiponectin in the epididymal fat were performed by real-time quantitative PCR.The relative contents of TNF-?,IL-6 and MCP-1 in peripheral adipose tissue were determined by ELISA.RESULT: The Lee's index of fat-1 transgenic mice was significantly lower than that of wild-type mice(P <0.05).The autophagosome of arcuate nucleus in fat-1 transgenic mice were more than those in wild type mice,while the expression of autophagy-related protein ATG7 was significantly up-regulated(P <0.05),and the proportion of LC3? / ? was significantly increased(P <0.05),and the expression of autophagy-related protein P62 was significantly decreased(P < 0.05)in fat-1 transgenic mice;The m RNA expression of TNF-?,IL-6,IL-1?,NF-k B,MCP-1,CCL5,CXCL12,TMEM119,GFAP in the hypothalamus of fat-1 transgenic mice was significantly higher than that in the control group(P <0.05),while the expression of CX3CL1 was up-regulated(P <0.05).The relative expression levels of TNF-?,IL-6,IL-1?,IFN-?,MCP-1 and TLR-4 in fat-1 transgenic mice were down regulated(P<0.05),while the relative expression of adiponectin in the transgenic mice was significantly increased(P<0.05).The protein content of TNF-?,IL-6 and MCP-1 in peripheral adipose tissue of fat-1 transgenic mice was significantly lower than that of wild type mice,that is,the inflammatory in peripheral adipose tissue of fat-1 transgenic mice was lower than that of wild type mice.CONCLUSION: Addition of n-3PUFAs in fat-1 transgenic mice,upregulation of hypothalamic autophagy,inhibition of the expression of hypothalamic inflammatory factors,chemokine expression,down-regulation of hypothalamic inflammation,maintenance of hypothalamic energy balance,thereby reducing body weight,prevention,prevention of obesity.