| Objective:By detecting the levels of lipids and lipid metabolism related genes and protein levels in rats exposed to different DEHP exposures,determine the effects of DEHP exposure on lipid metabolism in rats,by detecting the levels of inflammation and autophagy in.adipose tissue to investigate the role of autophagy and inflammatory response in lipid metabolism disorder induced by DEHP of rats,and provide scientific basis for prevention and treatment of lipid metabolism disorder.Method:Eighty healthy 21-day-old wistar rats were selected and randomly divided into 4 groups: control group(corn oil),low-dose DEHP group(5 mg/kg/d,1/6000 LD50),and medium-dose DEHP.Treated with 50 mg/kg/d,1/600 LD50 and high dose DEHP(500 mg/kg/d,1/60 LD50).Each group has 10 males and females.After 7 days of adaptive rearing,the mice were intoxicated once a day for 8 weeks.Weigh daily and record feeding conditions.Morphological changes of adipose tissue were observed by HE staining;IL-1β,TNF-α,LEP,and ADP in rat serum and adipose tissue were measured by ELISA;Serum TC,TG,HDL-C,and LDL-C were measured by automatic biochemical analyzer.Q-PCR method was used to determine the expression levels of lipid metabolism-related gene CEBP/β,inflammation-related gene CD68 and autophagy-related gene LC3 mRNA,and the protein expression was determined by Western Blot.Using Excel to enter data,SPSS 18.0 software was used for statistical analysis of experimental data,and normal data was expressed as x ±s.One-way analysis of variance(One Way ANOVA)was used to compare multiple groups.LSD or Kruskal-Wallis tests were used to compare the two groups.α=0.05 was used as the test level.Result:1.There was no significant difference in body weight between the rats in each group in the beginning 4 weeks(P<0.05).In the 5th to 8th week after exposure,the weight of rats in the high dose group was significantly higher than that in the control group(P<0.05).The rats in the high-dose group from the 2nd to the 3rd week of DEHP exposure had significantly higher food intake than the other groups(P<0.05).At the 4th week after the exposure,there was no significant difference in the feed intake between the rats in each group(P>0.05).);In the 5th to 8th weeks after exposure,the feed intake of rats in the high-dose group was significantly higher than that in the other groups(P<0.05);the food intake in the low-and middle-dose groups was significantly higher than that in the control group at the third week.(P<0.05),but in other weeks,there was no significant difference in food intake between the low and middle dose groups and the control group(P>0.05).2.Observed by microscope,compared with the control group,the amount of adipose tissue cells in the medium-dose and high-dose DEHP-treated groups increased,the sizes were irregular,the shape was irregular,and the macrophage infiltration increased.3.The levels of serum LDL-C,HDL-C,TC in rats exposed to high doses of DEHP were significantly higher than those in the control group(P<0.05);serum TG levels in rats increased slightly with increasing dose of DEHP,but there were no statistics between groups.Differences in learning(P>0.05);LEP levels in adipose tissue of rats exposed to medium and high doses of DEHP were significantly higher than those in the control and middle dose groups(P<0.05);ADP levels in adipose tissue in rats exposed to DEHP at each dose Significantly lower than the control group(P<0.05).4.The levels of IL-1β and TNF-α in adipose tissue of rats exposed to DEHP were significantly higher than those in the control group(P<0.05).The serum levels of IL-1β and TNF-α in rats exposed to high dose of DEHP were significantly higher.In the control group(P<0.05).5.The levels of CEBP/β,CD68,and LC3 mRNA expression in adipose tissue of rats exposed to DEHP were significantly higher than those in the control group;the expression levels of CEBP/β,CD68,and LC3 mRNA in adipose tissue of rats exposed to DEHP at medium and high doses All of them were significantly higher than low-dose group(P<0.05).The expression of CD68 mRNA in adipose tissue of high-dose DEHP group was significantly higher than that of middle-dose group(P<0.05).6.The expression levels of CEBP/β,CD68,and LC3II/LC3 I protein in adipose tissue of rats exposed to DEHP were significantly higher than those in the control group(P<0.05).The expression level of CEBP/β protein in DEHP treated group was higher than that in DEHP group.Control group,low-dose group and high-dose group(P<0.05);CD68 protein expression level increased gradually with increasing dose of DEHP;the difference was statistically significant(P < 0.05);middle-dose and high-dose DEHP-treated group LC3II/LC3 I levels were significantly higher than low-dose groups(P<0.05).7.In medium-and high-dose DEHP-treated groups,levels of serum TC and adipose tissue LEP and AD were significantly increased in adipose tissue in high IL-1β level subgroups(P<0.05),and high-level subgroups were found.Significantly higher than low-level group(P<0.05);In high-dose DEHP-treated group,serum HDL-C levels in rats with high IL-1β level were significantly higher(P<0.05).In the low-dose group,LEP levels in the adipose tissue of the high-level TNF-α subgroup were significantly higher than those in the low-level subgroup(P<0.05).In the medium-dose DEHP-poisoned group,the levels and high levels of TNF-α were significantly higher.Serum TC levels in subgroups were significantly higher than those in low-level subgroups(P<0.05),and higher levels in subgroups were higher than in low-level subgroups(P<0.05).LEP levels in adipose tissue in high-level TNF-alpha subgroups were higher than in low levels.In the medium-and intermediate-level subgroup(P<0.05),in the high-dose DEHP-infected group,serum TC,LDL-C,and adipose tissue LEP levels were significantly higher in the TNF-α and high-level subgroups than in the low-level subgroups.The group was higher than the mid-level subgroup(P<0.05).In the medium-dose DEHP exposure group,serum TC levels in high-level CD68 rats were significantly higher than in low-and medium-level subgroups.LEP tissue levels in high-level and subgroups of CD68 were higher than those in low-level subgroups,and high levels of subgroups.The group was higher than the mid-level subgroup(P<0.05).8.In low-dose and high-dose DEHP-treated groups,serum TG levels in the subgroup of rats with high levels of autophagy decreased significantly(P<0.05).In the middle-dose DEHP-treated group,levels of autophagy and high levels of The level of AD in adipose tissue of rats was significantly higher(P<0.05),and AD in high-level subgroup was higher than that in middle-level subgroup(P<0.05).Conclusion:1.DEHP exposure can affect the expression of lipids and lipid metabolism related genes and proteins in adolescent rats and affect the fat cell differentiation and adipose tissue morphology,interfere with feeding and weight gain in rats,resulting in lipid metabolism disorder in adolescent rats.2.DEHP exposure can cause different degrees of inflammatory conditions associated with macrophage infiltration in rat serum and adipose tissue.3.Exposure to DEHP induces increased levels of autophagy in adipose tissue.4.Adipose tissue inflammation and autophagy may play a role in lipid metabolism disorder in adolescent rats induced by DEHP exposure. |
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