| Objective: This research plan uses transgenic mice and electrical stimulation C2C12 cell models to deeply study the role of lipid droplet protein 17?-HSD11 in the regulation of skeletal muscle lipid droplet metabolism and insulin resistance in electrical stimulation simulation exercise,and reveal its related molecular mechanism.Exploring the molecular mechanism of lipid metabolism regulation provides valuable clues for studying the relationship between exercise and health.Methods: Select 6 8-week-old C57BL/6 wild-type male mice with uniform body weight and inject anesthetic.After the rats were fully anesthetized,their tissues were quickly taken out for real-time quantitative PCR experiment.Furthermore,the purified lipid drops of skeletal muscle,liver and brown fat were used for Western blot analysis to analyze the expression and localization of 17?-HSD11.The piggy Bac technology was used to construct transgenic mice overexpressing 17?-HSD11,and then 12 male mice with uniform litter weight and the same age were selected from the animal room and divided into wild type group(C)and transgenic group(H);record their weights,Carry out the glucose tolerance test;Put it into the metabolic cage to collect the respiratory quotient data;After completion,use prism software for data analysis.Then the mouse tissues were taken,prepare samples and use Western Blot and real-time quantitative PCR to analyze the effect of 17?-HSD11 on lipid droplet-related proteins influences.A triglyceride kit was used to measure triglyceride content in skeletal muscle and serum.Mouse myoblasts C2C12 were cultured in a six-well plate and induced to differentiate into mature skeletal muscle polynucleated myotube cells.They were divided into 6 groups: control group(C),palmitic acid treatment group(PA),oil Acid treatment group(OA),electrical stimulation group(ES),electrical stimulation and palmitic acid co-treatment group(ES+PA),electrical stimulation and oleic acid co-treatment group(ES+OA).After 40 V,2ms,1Hz electrical stimulation,500?M palmitic acid and200?M oleic acid intervention,Western blot was used to determine the protein expression of ADRP,p-Akt,AKT and 17?-HSD11 in mouse skeletal muscle myotube cells C2C12.Results:(1)17?-HSD11 widely expressed in the body.(2)17?-HSD11 was localized to the surface of lipid droplets.(3)17?-HSD11 in the expression of skeletal muscle lipid droplets compared with liver lipid droplets expression,17?-HSD11 expressed in skeletal muscle droplets inferior to levels.(4)There was no variance in the blood sugar values between WT and overexpressed mice at each time point after the injection of glucose.(5)Respiratory entropy of WT mice was prominently inferior to that of overexpressed mice(P<0.05).(6)The triglyceride content of WT mice was prominently inferior to that of overexpressed mice(P<0.05).(7)The utterance levels of ATGL and OXPAT in overexpressed mice were prominently above prep those in WT mice(P<0.05).The utterance level of ADRP in overexpressed mice was above prep that in WT mice,and the utterance level of GLUT4 in overexpressed mice was inferior to that in WT mice,but there was no prominent difference.(8)The utterance levels of ADRP,ATGL and OXPAT in PA and OA groups were prominently above prep those in C group.(9)The utterance level of p-Akt in PA group was inferior to that in C group,and there was no prominent difference in p-Akt utterance between OA group and C group.(10)The utterance levels of ATGL and OXPAT in ES group were prominently above prep those in C group,but the utterance levels of ADRP were not prominently variant.The utterance level of ADRP in ES+PA group was inferior to that in PA group,and the utterance level of ADRP in ES+OA group was inferior to that in OA group.The utterance level of ATGL m RNA in ES+OA group was above prep that in OA group.(11)The utterance levels of p-Akt and GLUT4 in ES+PA group were above prep those in PA group.Conclusions:(1)17?-HSD11 expressed in generalized body,localized in lipid droplets,and expressed in skeletal muscle lipids,but lower than that in liver lipids.(2)17?-HSD11 affect the energy metabolism of mice to make the body nutrition surplus,increase the accumulation of the skeletal muscle triglyceride,induce ectopic lipid storage;17?-HSD11 affects the synthesis and decomposition of lipid droplets and affects the expression of GLUT4.(3)Fatty acids improved lipid droplets content in mouse skeletal muscle cells by affecting lipid droplets surface proteins.Palmitic acid decreased p-Akt utterance and induced insulin resistance in mouse skeletal muscle C2C12 cells.Oleic acid had no effect on p-Akt utterance.Electrical stimulation simulates the effects of aerobic exercise to promote the utterance of ATGL and OXPAT,reduce lipid accumulation,addition p-Akt and GLUT4 utterance,and improve insulin resistance. |
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