| Objective To establish insulin-resistant L02 cells with high concentration insulin.To explore the effect of rosiglitazone on the expression of ANGPTL3 and LXRαin normal and Insulin-Resistant human liver cells in different concentration of glucose states.To investigate the relation between ANGPTL3,LXRαand the diabetes mellitus,metabolic syndrome . Western blotting was used to detect the difference of ANGPTL3 serum content of the normal and the patients with hyperglycaemia,hyperglycaemia and high cholesterol,hyperglycaemia and high triglyceride .To explore the effect of Lipid and glucose on the ANGPTL3 serum content. The detecting of ANGPTL3 can lay the foundation for analyzing its relationship with related diseases.Methods①L02 cells were used as research object.The cells were devided into 4 groups: Insulin-resistant group(IR),Insulin-resistant which were added rosiglitazone group(IR-R),normal group(N),normal which were added rosiglitazone group(N-R).every group was cultured in mediums containing different concentration of glucose which were 5.6,7.0,11.1, 33.0mmol/L. The content of glucose remained in the culture solution was measured by the method of glucose oxidizes peroxides(GOD-POD),and this method was used to judge the Insulin-resistant L02 cells. The expression of Angptl3 mRNA and LXRαmRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR).②Clinical serum specimens(normal blood glucose and blood-fat group 7 Cases,hyperglycaemia group 6 Cases,hyperglycaemia and high cholesterol group 5 Cases,hyperglycaemia and high triglyeride group 6 Cases) were collected.The difference of ANGPTL3 serum content was detected by western blotting.Results①The content of glucose remained in culture solution of IR-R group was lower than the group of IR group(P<0.05).②The increasing of glucose content stimulated the expression of ANGPTL3 in N group,N-R group,IR group and IR-R group. In the same glucose content environment , the expression of ANGPTL3 of N1 group and IR1 group has no obvious difference(P>0.05),while ANGPTL3 expression of IR2- IR4 group were higher than N2～N4 group(P<0.05). In the same glucose content environment ,ANGPTL3 expression of IR-R group were higher than IR group(P<0.05)and the expression of ANGPTL3 of N-R group were higher than IR-R group(P<0.05).The expression of LXRαof N 1,N-R1,IR1 and IR-R1 have no obvious difference with N 2,N-R2,IR2 and IR-R2(P>0.05),while the expression of LXRαof N 2～4,N-R2～4,IR2～4 and IR-R2～4 have obvious difference in each group(P<0.05).While the glucose content was higher than 11.1mmol/L, the glucose content can stimulate the expression of LXRα,and the expression of LXRαwas increasing with the glucose content added (P<0.05). In the same glucose content environment , the expression of LXRαof IR group was lower than N group(P<0.05)and the expression of LXRαof IR-R group was higher than N group(P<0.05),the expression of LXRαof N-R group was higher than IR-R group(P<0.05).③ANGPTL3 serum content of hyperglycaemia group was higher than normal blood glucose and Lipids group(P<0.05), ANGPTL3 serum content of hyperglycaemia and high Lipids group was higher than hyperglycaemia group and normal group. ANGPTL3 serum content of hyperglycaemia and high cholesterol group was higher than hyperglycaemia and high triglyeride group,but they have no obvious difference in statistics(P>0.05).Conclusions 1.Insulin-resistant L02 cells model was established Successfully.2.Rosiglitazone can improve Insulin-resistant states of cells and increase the expression of LXRα.The increasing of the glucose intaked and high expression of LXRαcan stimulate the expression of Angptl3,and the increasing expression of Angptl3 was higher than the decreasing expression of Angptl3 by the improving of Insulin-resistant states. 3. ANGPTL3 serum content was higher in hyperglycaemia group and hyperglycaemia and high Lipids group than in the normal group.The result can reveal that ANGPTL3 serum content has relationship with those disease. |
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