| Objective:To describe the effect of the action time of inducers on the differentiation of 3T3-L1 cells to adipocytes.Methods:According to the "cocktail" method,three groups divided by the action time of inducers were involved into the study,with the action time of 2,3 or 4 days,respectively.Cell morphology was observed using inverted microscope and adipose content were detected by Oil red O staining and ELISA.Results:The differentiation rate in 2-day group was significantly lower than that in 3-or 4-day groups(65%vs.100%,P<0.05).As for the adipose content,the value in 4-day group was significantly higher than that in 2-or 3-day group(1402.5±9.4 vs.1078.5±15.2 或 1083.4±9,6 ng/mL,P<0.05),while no difference between the 2-and 3,day groups(1078.5±15.2 vs.1083.4±9.6 ng/mL,P>0.05).Conclusions:Improved procedure for differentiation of 3T3-L1 cells to adipocytes could deserve one of the methods to made adipocyte models.Preferentially,the procedure with its action time of the inducers of 3 days was recommended.Objective The purpose of this study is to investigate the effect of 3T3-L1 on the function of human umbilical vein endothelial cells under the stimulation of hyperglycemia.Methods Human umbilical vein endothelial cells were selected as the research object,and the experiment was divided into 3 groups:normal control group(A),high glucose stimulation group(B),high glucose stimulation group cocultured with 3T3-L1 adipocytes(C).With Tubilin as control,the expression of ICAM-1,OPN,and eNOS in human umbilical vein endothelial cells was detected using methods of Western blot and rt-PCR,respectively.Results The expression of eNOS in group B was significantly lower than that in group A and C(P<0.05),and the expression of eNOS in group C was also significantly lower than that in group A(P<0.05).On the contrary,the expression of ICAM-1 and OPN in group B was significantly higher than that in group A and group C(P<0.05),but there was no significant difference between the group A and group C(P>0.05).Conclusions Hyperglycemia can lead to dysfunction of human umbilical vein endothelial cells.3T3-L1 adipocytes could alleviate the damage of human umbilical vein endothelial cells caused by hyperglycemia through decreasing the expression of eNOS,ICAM-1 and OPN.Objective:To investigate the role of perivascular adipose tissue in the repair of injured endothelium in diabetic rats.Methods:Male SD rats were selected as the research object and the study were randomly divided into 3 groups:control group(group A),diabetes mellitus + carotid artery balloon injury group(group B),diabetes mellitus + carotid artery balloon injury + perivascular adipose tissue transplantation group(group C).The expression of ICAM-1,OPN,eNOS were all detected using Western Blot,rt-PCR and immunohistochemical method and,the severity of intimal proliferation was also observed by Hematoxylin-eosin(HE)staining.Results:The results of HE staining showed that the thickness of carotid artery in B group was significantly higher than that in group A and C(p<0.05),but no significant difference between C group and A group was found(p>0.05).The results of Western,rt-PCR and immunohistochemistry showed that the expression of eNOS was significantly lower in B group than that in A group and C group(p<0.05),while the expression of ICAM-1 and OPN was significantly higher than that of A and C two(blot)group(p<0.05).Conclusion:Perivascular adipose tissue can promote the recovery of injured endothelium in diabetic rats and,ICAM-1 and OPN may be the potential therapeutic target for diabetic vascular disease. |
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