The Effect Of TSH On The Expression Of Adipose Cell’s Inflammatory Factors And Its Mechanism Study

ObjectiveObserve the influence of TSH to inflammation factors above TNF-α and adiponectin,which secreted by adipoeyte eell.At the same time to explore its mechanism.Preliminary study of the correlation between TSH and atherosclerosis.1.3T3-L1 preadipocyte differentiate into mature adipocyte in vitro3T3-L1 preadipose cell cultivated in vitro,and through the classic cocktails induced into mature adipose cells.For the study of TSH in mature adipose cells laid a solid foundation for the mechanism of action of inflammatory cytokines.2.The influence of TSH to TNF-α secretion from mature adipocyteUsing TSH to stimulate mature adipose cell in different times,and by ELISA to detect the level of TNF-α.Identified the time as the following experiments which TSH stimulates to mature adipose cell.Then,with different concentrations of TSH(0.0 μM、0.1 μM、1μM),1μM cAMP agonist(Forskolin),and 10μM protein kinase inhibitors(H89)which intervents in 15 minutes and then using 1μM TSH to stimulates mature adipose cell,respectively,to detect the concentration of TNF-α,the expression of NF-κ B p65 and the change of TNF-α mRNA.3.The influence of TSH to ADPN secretion from mature adipocyteUsing TSH to stimulate mature adipose cell in different times,and by ELISA to detect the level of ADPN.Identified the time as the following experiments which TSH stimulates to mature adipose cell.Using different concentration of TSH(0.01μM、0.1μM、1μM)to stimulate mature adipose cell,and detecting the concentration of ADPN,the expression of PPAR Y and the change of ADPN mRNA.Result1.3T3-L1 preadipocyte cells were successfully induced into mature adipocyte cells.And identified by oil red O staining.2.As the TSH stimulation time extended,differentiation of mature 3T3-LI adipose cell in the culture medium of INF-α concentration is on the rise,stimulate the 4 h to reach peak(P<0.05).Compared with the PBS group,with the increase of the concentration of TSH stimulation,the medium level of INF-a,TNF-a mRNA level is on the rise in the cell(P<0.05),the NF-K B P65 protein expression level is on the rise.Compared with 1μM TSH stimulation group,Forskolin medium level of TNF-a stimulating group no significant change(P>0.05);Compared with the PBS group,medium level of TNF-α Forskolin stimulation group was obviously increased(P<0.05).In PBS group,compared to 10MM H89 group of TNF-α level not seen in culture medium(P>0.05).Preliminary discussion on TSH through the cAMP-PKA pathways,activate the NF-κB,then promote the secretion of 3T3-LI adipose cell that TNF-α,participate in the occurrence of atherosclerosis.3,With the increase of concentration of TSH stimulation,reduced the secretion of ADPN significantly,PPAR Y,ADPN mRNA expression is also reduced.Conclusion1.Through the cAMP-PKA,TSH activates the NF-κB and promotes the secretion of TNF-α,at the same time,participate in the oeeurence of atherosclerosis.2.TSH induced the secretion and expression of ADPN and PPAR γ.