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Expression of extracellular superoxide dismutase during adipose differentiation in 3T3-L1 cells
Tetsuo Adachi1, Taisuke Toishi, Haoshu Wu
1Laboratory of Clinical Pharmaceutics, Gifu Pharmaceutical University, Gifu, Japan. adachi@gifu-pu.ac.jp
Abstract:
Obesity is known to be the primary causal component in metabolic syndrome. Adipocytes in obese patients exhibit increased oxidative stress via the activation of reactive oxygen species (ROS)-producing systems and inactivation of antioxidant enzymes. Extracellular superoxide dismutase (EC-SOD) is an anti-inflammatory enzyme that protects cells from the damaging effects of ROS. An earlier report showed that plasma EC-SOD levels in type 2 diabetic patients were significantly and inversely related to body mass index and homeostasis model assessment-insulin resistance index. Moreover, the administration of pioglitazone, an antidiabetic agent, significantly increased the plasma level of EC-SOD. In this report, the expression of EC-SOD was compared to other adipocytokines in mice 3T3-L1 pre-adipocytes. EC-SOD expression levels were increased after the induction of differentiation and then declined, which was similar to adiponectin and transcription factors such as peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and CCAAT/enhancer-binding protein-alpha (C/EBP-alpha). On the other hand, the expression levels of pro-inflammatory adipocytokines, such as tumor necrosis factor-alpha (TNF-alpha) and monocyte chemo-attractant protein-1 (MCP-1), increased markedly in the development stage of cells. It was observed that the expression of EC-SOD in differentiated 3T3-L1 cells co-cultured with LPS-stimulated J774 macrophages was up-regulated, while the addition of TNF-alpha down-regulated EC-SOD and adiponectin expression in adipocytes. It is known that infiltrated and activated macrophages produce extracellular ROS at high levels in adipose tissue. It is possible that the expression of EC-SOD in adipocytes was stimulated to protect them from oxidative stress in the co-culture system.
Insights
Obesity-induced oxidative stress impacts metabolic syndrome. This study reveals extracellular superoxide dismutase (EC-SOD) expression in adipocytes, potentially protecting against ROS and inflammation.
Area of Science:
- Biochemistry
- Cell Biology
- Metabolic Syndrome Research
Background:
- Obesity is a primary driver of metabolic syndrome, characterized by increased oxidative stress in adipocytes due to ROS-producing systems and reduced antioxidant enzyme activity.
- Extracellular superoxide dismutase (EC-SOD) is an anti-inflammatory enzyme that combats ROS damage. Previous studies link lower plasma EC-SOD to higher BMI and insulin resistance, with pioglitazone increasing EC-SOD levels.
Purpose of the Study:
- To compare the expression of EC-SOD with other adipocytokines during the differentiation of mice 3T3-L1 pre-adipocytes.
- To investigate the regulatory role of macrophages and pro-inflammatory cytokines on EC-SOD expression in adipocytes.
Main Methods:
- 3T3-L1 pre-adipocytes were induced to differentiate, and EC-SOD expression was analyzed alongside adiponectin, PPAR-gamma, C/EBP-alpha, TNF-alpha, and MCP-1.
- Differentiated 3T3-L1 cells were co-cultured with LPS-stimulated J774 macrophages, and the effects of TNF-alpha on EC-SOD and adiponectin expression were assessed.
Main Results:
- EC-SOD expression initially increased with differentiation, similar to adiponectin and key transcription factors, but later declined.
- Pro-inflammatory adipocytokines (TNF-alpha, MCP-1) showed increased expression during adipocyte development.
- Co-culture with macrophages upregulated EC-SOD in adipocytes, while TNF-alpha downregulated EC-SOD and adiponectin expression.
Conclusions:
- EC-SOD expression in adipocytes follows a differentiation-dependent pattern, mirroring adiponectin and key adipogenic transcription factors.
- Macrophages and their secreted factors, like TNF-alpha, significantly influence EC-SOD expression in adipocytes.
- EC-SOD upregulation in response to macrophage co-culture suggests a protective mechanism against ROS-induced oxidative stress in adipose tissue during obesity.
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