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Abstract
Aquaglyceroporins, belonging to the family of aquaporins (AQPs), are integral plasma membrane proteins permeable to water and glycerol that have emerged as key players in obesity. The aim of this study was to investigate the expression profile of AQPs in undifferentiated and differentiated 3T3-L1 cells and to investigate the changes in expression of aquaglyceroporins in 3T3-L1 cells differentiated into adipocytes and subjected to lipopolysaccharide (LPS) mimicking inflammation occurring during obesity. Furthermore, the study aimed at identifying the signaling cascade involved in the regulation of aquaglyceroporins expression upon LPS stimulation. 3T3-L1 cells were grown as undifferentiated cells (UDC; preadipocytes) or cells differentiated into adipocytes (DC, adipocytes). DC were incubated in the presence or absence of LPS with or without inhibitors of various protein kinases. AQPs mRNA expression levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). AQP1, AQP2, AQP3, AQP9 and AQP11 mRNA were expressed in both UDC and DC, whereas AQP4, AQP7 and AQP8 mRNA were expressed only in DC. In DC, LPS up-regulated AQP3 mRNA levels (p < 0.05) compared to control; these effects were inhibited by CLI095, SP600125 and BAY11-7082 (p < 0.05). LPS decreased both AQP7 and AQP11 mRNA levels (p < 0.01) in DC as compared to control; this decrease was inhibited by CLI095 and BAY11-7082 (p < 0.05) and additionally by SP00125 for AQP7 (p < 0.05). SB203580 had no effect on LPS-induced AQP3, AQP7 and AQP11 mRNA levels modulations. In conclusion, our results clearly show that many AQPs are expressed in murine 3T3-L1 adipocytes. Moreover, in DCs, LPS led to decreased AQP7 and AQP11 mRNA levels but to increased AQP3 mRNA levels, resulting from the Toll-like receptor 4 (TLR4)-induced activation of JNK and/or NFκB pathway.
Highlights
The incidence of obesity worldwide is increasing constantly and represents a critical risk factor for the onset of various diseases such as cardiovascular diseases, diabetes mellitus, hyperlipidemia and cardiac infarction [1,2]
The aim of this study was to determine the AQPs expression profile in undifferentiated 3T3-L1 cells (UDC; preadipocytes) vs. 3T3-L1 cells differentiated into adipocytes (DC; adipocytes), as well as the modulation of aquaglyceroporins expression by LPS in the DC
Expression of Aquaporins (AQPs) mRNA in 3T3-L1 Cells Differentiated into Adipocytes (DC; Adipocytes) vs. Undifferentiated 3T3-L1 Cells (UDC; Preadipocytes)
Summary
The incidence of obesity worldwide is increasing constantly and represents a critical risk factor for the onset of various diseases such as cardiovascular diseases, diabetes mellitus, hyperlipidemia and cardiac infarction [1,2]. The increase of adipose tissue mass that accompanies obesity is due to an expansion in adipocyte number (hyperplasia) and size (hypertrophy) [3]. Adipocytes play a crucial role in whole-body energy homeostasis as it supplies energy during starvation. Glycerol plays a major role in energy homeostasis, as it represents the carbon backbone of triacylglycerol (TAG) and constitutes the major substrate for hepatic gluconeogenesis during fasting [5,6]. Circulating plasma glycerol results from its release from TAG during lipolysis and its reabsorption by kidney proximal tubules [7,8]
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