[Effects and mechanism of testosterone on the production of inflammatory cytokines and glucose uptake in co-culture of RAW264.7 macrophage and 3T3-L1 adipocytes].
Objective:To investigate the effects of testosterone (T) on inflammatory cytokines (IL-6, MCP-1) production, insulin sensitivity of adipocyte and changes of macrophage phenotypes in indirect co-culture of RAW264.7 macrophages and 3T3-L1 adipocytes.Methods:3T3-L1 preadipocytes were induced to mature in Transwell lower chamber, and then co-cultured with RAW264.7 macrophages in the upper chambers for 72 hours. Testosterone 10 μmol/L was added into indirect co-culture for 24 h. ELISA was used to testing IL-6, MCP-1 concentrations in supernatant. Western blot was used to detecting the phosphorylation of NF kappa B, ERK1/2, and theexpression of CD16/32 and CD206. Glucose transport was assessed by[3H]2-deoxy glucose uptake in adipocytes.Results:Testosterone enhanced inflammatory cytokines (IL-6, MCP-1) production in indirect co-culture of 3T3-L1 adipocytes and RAW264.7 macrophages, promoted the activation of ERK1/2 and nuclear factor kappa B p65, and inhibited glucose uptake in adipocytes. Testosterone facilitated the production of pro-inflammatory M1macrophages. The above effects of testosterone can be completely reversed by PDTC, and can be partly reversed by PD98059 (70%-90%).Conclusion:NF kappa B and ERK1/2 could be the key proteins for testosterone to promote the production of inflammatory factors, to lead to insulin resistance, and to make macrophages differentiate to pro-inflammatory phenotypes in co-culture of RAW264.7 macrophages and 3T3-L1 adipocytes.
Obstetrics and Gynecology Hospital Affiliated to Fudan University, Shanghai 200011, China.
C L
CL
Su
M
M
Chen
P N
PN
Zhang
W
W
Xu
J F
JF
Lin
D041721:3T3-L1 Cells; D017667:Adipocytes; D000818:Animals; D018920:Coculture Techniques; D016207:Cytokines; D005947:Glucose; D007333:Insulin Resistance; D008264:Macrophages; D051379:Mice; D048052:Mitogen-Activated Protein Kinase 3; D016328:NF-kappa B; D010766:Phosphorylation; D000067996:RAW 264.7 Cells; D013739:Testosterone
D016207:Cytokines; D016328:NF-kappa B; D013739:Testosterone; D048052:Mitogen-Activated Protein Kinase 3; D005947:Glucose