Effects Of Adipose Tissue From Different Regional On Glucose Uptake And Glycogen Synthesis In Hepatocyte

Objective:To investigate the effects of different regional of adipose tissue(AT)on glucose uptake and glycogen synthesis in hepatocytes.Methods:Human HepG2 cells,subcutaneous adipose tissue(SAT)and visceral adipose tissue(VAT)were cultured in groups,and the effects of SAT and VAT on glucose uptake and glycogen synthesis of hepatocytes were detected respectively.The hepatocytes were co-cultured in Transwell chamber in vitro,and the supernatant of each group was detected by LDH method after co-culture to determine the activity of hepatocytes.After co-culture,HepG2+VAT co-culture group and HepG2+SAT co-culture group were incubated in the medium with or without insulin for 30 minutes,respectively.The cells were collected after incubation at 37?.Glucose uptake into the cells was detected by 2-NBDG(2-NBDG fluorescent glucose analogue)method,and the proportion of fluorescent labeled cells was measured by flow cytometry.The synthesis of glycogen in cells was measured by anthrone method.Results:(1)Glucose uptake:there was significant difference in glucose uptake of HepG2 cells among HepG2 group,HepG2+SAT group and HepG2+VAT group,(HepG2 group>HepG2+SAT group>HepG2+VAT group:99.60±0.14>56.35±0.83>51.12±1.32(P<0.001)).Compared with HepG2 group,the decrease of glucose uptake of HepG2 cells in HepG2+VAT group was more obvious than that in HepG2+SAT group:GU%=-48.49±1.27<-43.26±0.84(P<0.001).There was significant difference in glucose uptake of HepG2 cells among HepG2+INS group,HepG2+SAT+INS group and HepG2+VAT+INS group,(HepG2+INS group>HepG2+SAT+INS group>HepG2+VAT+INS group:98.57±0.55>77.15±1.18>70.57±1.48(P<0.001)),and compared with HepG2+INS group,the decrease of glucose uptake of HepG2 cells in HepG2+VAT+INS group was more obvious than that of HepG2 cells in HepG2+SAT+INS group:GU%=-28.01 ± 1.96<-21.42 ± 1.52(P<0.001).(2)Glycogen synthesis:compared with HepG2 group,HepG2+SAT group and HepG2 group,the difference of glycogen synthesis of HepG2 cells was statistically significant,(HepG2 group>HepG2+SAT group>HepG2+VAT group:5.00 × 10-2±0.31 × 10-2>2.44 × 10-2 10.36 × 10-2>1.13 × 10-2 10.23 × 10-2(P<0.001)).Compared with HepG2 group,the glycogen synthesis of HepG2 cells in HepG2+VAT group and HepG2+SAT group decreased more significantly than that in SAT group:GS%:-77.38 ±4.21<-50.75 19.53(P<0.001):Compared with HepG2+INS group,HepG2+SAT+INS group and HepG2+VAT+INS group,the difference of glycogen synthesis of HepG2 cells was statistically significant:(HepG2+INS group>HepG2+SAT+INS group>HepG2+VAT+INS group:4.15 ×10-2±0.29 × 10-2>3.11 × 10-2 10.26 × 10-2>2.22 ×10-2 10.37 × 10-2(P<0.001));compared with HepG2+INS group,the decrease of glycogen synthesis of HepG2 cells in HepG2+VAT+INS group was more obvious than that in HepG2+SAT+INS group:GS%:-46.38±9.38<-24.88±5.5 1(P<0.001).conclusion(1)The glucose uptake of HepG2 cells co-cultured with human SAT and VAT was lower than HepG2 cells cultured alone,and the decrease of glucose uptake of HepG2 cells co-cultured with VAT was more obvious,suggesting that both VAT and SAT could inhibit glucose uptake in hepatocytes,and the inhibitory effect of VAT on glucose uptake of hepatocytes was more significant than that of SAT.(2)Glycogen synthesis:the amount of glycogen synthesis of HepG2 cells co-cultured with human SAT and VAT was lower than HepG2 cells cultured alone,and the decrease of glycogen synthesis of HepG2 cells co-cultured with VAT was more obvious,suggesting that both VAT and SAT could inhibit glycogen synthesis in hepatocytes,and the inhibitory effect of VAT on glycogen synthesis of hepatocytes was more significant than that of SAT.