Premium
Studies on the cell treatment conditions to elicit lipolytic responses from 3T3‐L1 adipocytes to TCDD, 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin
Author(s) -
Li Wen,
Vogel Christoph F.A.,
Matsumura Fumio
Publication year - 2007
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.21303
Subject(s) - lipolysis , medicine , endocrinology , adipose tissue , adipocyte , 3t3 l1 , incubation , downregulation and upregulation , tetrachlorodibenzo p dioxin , chemistry , in vivo , biology , toxicity , biochemistry , microbiology and biotechnology , gene
Abstract Wasting syndrome is one of the hallmark symptoms of poisoning by TCDD (=dioxin), which is associated with the massive loss of adipose tissue and serum hyperlipidemia in vivo. Yet, the most widely used in vitro cell model 3T3‐L1 adipocyte has not been useful for studying such an action of TCDD because of the difficulty of inducing their mature adipocytes to respond to TCDD to go through lipolysis. Here, we made efforts to find the right cell culture and treatment conditions to induce mature 3T3‐L1 adipocytes to go through lipolysis, which is defined as events leading to reduction of lipids in adipocytes. The optimum condition was found to require 7‐day differentiated adipocytes being subjected to DMEM medium containing TCDD (but without insulin) for 5 day incubation with two medium changes (the same composition) on incubation days 2 and 4. After 24 h, the early effect of TCDD on adipocytes was predominantly on inflammation, particularly induction of COX‐2 and KC (IL‐8), which is accompanied by upregulation of C/EBPβ and δ. The sign of TCDD‐induced lipolysis starts slowly and by incubation day 3, a few markers showed modestly significant changes. By day 5 of incubation, however, many markers show highly significant signs of lipolytic changes. Although this process could take place without exogenous macrophages or their cytokines, addition of exogenous TNFα considerably synergized this action of TCDD. In conclusion, under a right condition, 3T3‐L1 adipocytes were found to respond to TCDD to go through lipolysis. The early trigger of such a response appears to be activation of COX‐2, which is amplified by TNFα. J. Cell. Biochem. 102: 389–402, 2007. © 2007 Wiley‐Liss, Inc.