Effects Of Eicosapentaenoic Acid On Inflammatory Factor,Blood Glucose,Blood Lipid And Insulin Resistence In Gestational Diabetic Mice

Objectives To study the effect of eicosapentaenoic acid on glucolipid metabolism and insulin resistance in gestational diabetes mice,and to explore initate mechanism of effect of EPA on insulin resistance by reducing the levels of inflammatory factors.Methods Thirty-two C57/BL mice that were fed high-fat diet for four weeks and were newly pregnant were selected,and 12 normal control（NC）mice were selected from the normal diet pregnant mice.32 mice were on the 5th,6th,and 7th day of pregnancy for three consecutive days the GDM model mice was established by injecting 50mg/Kg·bw of STZ solution;three days of abdominal blood glucose greater than 11.1mmol/L were GDM.The GDMs that were finally modeled and survived the experiment were randomly divided into three:GDM disease model,low-concentration EPA（100mg/Kg·bw）intervention groups,and high-concentration EPA（300mg/Kg·bw）intervention groups.On the 7th to 18th day of pregnancy,the two intervention ways were given two doses of EPA by gavage each day,and the other two were given the same volume of solvent by gavage.The daily blood glucose of pregnant mouses during pregnancy was recorded,and the mouses were sacrificed at GD18.The retained serum measures fasting blood glucose by blood sugar-test paper,blood lipids and insulin levels by ELISA kit;WB detects IL-6,IL1-β,and TNF-αprotein levels in the placenta,and immunohistochemical methods detect the expression of TLR4 on the surface of the placenta.HE and PAS staining were used to analyze the fatty degeneration and glycogen content in the liver of pregnant mice;Western blot was applied to analyze the expression levels of p IRS1Tyr,IRS1,GLUT4,p AKT2Ser,and AKT2 in the muscle tissue all gruop.Results 1 Comparison of fasting plasma glucose,insulin level and insulin sensitivity index（ISI）of gestational mouses among the four.The fasting plasma glucose of gestational mouses in the GDM was higher than that in the NC,and the fasting plasma glucose of gestational mouses in the GDM+HI was significantly lower than that of the GDM.The blood glucose level in the GDM+HI was slightly lower than that in the GDM+LI.Compared with NC,the FINS level of gestational mouses in the GDM was significantly higher.The FINS level of gestational mouses in the GDM+LI was lower than that in the GDM,while the FINS level of GDM+HI decline.The difference was statistically significant;compared with NC,The ISI level of gestational mouse in the GDM was significantly reduced,and the ISI of the gestational mouse in the GDM+LI was higher than that in the GDM,while the ISI level of the GDM+HI was higher than that of the GDM+LI.The above deviation were statistically significant.2 Comparison of the expression content of TC,TG,HDL,and LDL in pregnant mice serum among the fours:Compared with NC,TC,TG,LDL in GDM were significantly increased,while HDL was significantly reduced;TC,TG and LDL in GDM+HI,there is a downward trend compared with the GDM.The HDL expression content has risen.Compared with the GDM+LI,the GDM+HI had higher HDL expression content,while the LDL,TG,and TC contents decreased,and the difference was statistically significant.3 Comparison of HE staining in the hepatic tissue of fours on gestational mouse:Compared with NC,the proportion of vacuolar degeneration of hepatocytes in the hepatic tissue of gestational mouse in the GDM was significantly increased,and the difference was statistically significant.The proportion of vacuolar hepatocytes in the hepatic tissue of mouses in the GDM+LI was significantly decline compared with that in the GDM.In the GDM+HI,compared with the GDM+LI,the ratio of vacuole-like changes in the hepatocytes of the former was declined,and the deviation was statistically significant.4 The PAS staining of liver glycogen in the four of pregnant mouses:Compared with NC,the glycogen content of pregnant mouses in the GDM was significantly lower,and the differences were statistically significant.The glycogen content in the hepatic tissue of pregnant mouses in the GDM+LI was higher than that in the GDM.Compared with the GDM+LI,the glycogen content in the GDM+HI further increased,and the deviation were statistically significant.5 Comparison of the area under the curve of the OGTT results of the four of pregnant mouses:Compared with NC,the area under the curve of pregnant mouses in the GDM increased significantly,and the deviation were statistically significant.The glycogen content in the liver of pregnant mouses in the GDM+LI was lower than that in the GDM,while the area under the curve decreased in the GDM+HI compared with the GDM+LI,and the differences were statistically significant.6 Changes in the expression levels of key proteins in the insulin signaling pathway in the placenta of the fours of pregnant mice:Compared with NC,the expression levels of p IRS-1Tyr,GLUT4 and p AKTSer in the GDM were significantly reduced.Compared with GDM+LI and GDM,the expression of the three increased,and the difference was statistically significant.Compared with the GDM+LI,GDM+HI has an upward trend,and the difference is statistically significant.7 Comparison of Western blot results of the inflammatory factors IL-6,TNF-α,IL1-βin the placenta tissue of each of pregnant mouses:Compared with the NC,the relative expression contents of IL-1βand IL-6 in the placenta tissue of the GDM increased significantly;The relative expression contents of TNF-αincreased slightly.Compared with the GDM,the levels of IL-1β,IL-6 and TNF-αin GDM+HI pregnant mice decreased.Compared with the GDM+LI,the levels of IL-1β,IL-6,and TNF-αin pregnant mouses with GDM+HI decreased,and the above deviations were statistically significant.8 Comparison of the relative contens of TLR4 and NF-k B in the placenta of the four of gestational mouse:The results of immunohistochemistry and Western blot showed that the relative contents of TLR4 and NF-k B in the placenta of GDM mouse were significantly higher than those in the NC.Compared with the GDM,the relative contents of the two factors in the GDM+LI decline.Compared with the GDM+LI,the content of TLR4 and NF-k B in the GDM+HI decreased,and the deviation was statistically significant.9 Results of dual intervention of EPA and LPS:The relative expression contents of TLR4 and NF-k B in GDM+EPA+LPS were higher than those in the GDM+EPA and lower than those in the GDM+LPS.The deviation was statistically significant.The relative expression contents of p IRS1^Tyr in the GDM+EPA+LPS was higher than that in the GDM+LPS,but it was significantly lower than the relative expression contents of p IRS1^Tyrin the GDM+EPA.Conclusions 1 EPA can improve the state of glucose and lipid metabolism of GDM,and improve the degree of insulin resistance.2 EPA may inhibit the activation of TLR4 and blocking inflammatory reaction on the surface of the placenta tissue.Therefore,when the expression of inflammatory factors decreases,its inhibitory effect on insulin signaling is relieved.Figure 19;Table 14;Reference 130...