Abstract
There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at 42°C for one hour and then allowed to recover at normal incubation temperature of 37°C for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to 400 μg/mL) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.
Highlights
Heat stress imperils animal production efficiency through reduction of feed intake and diminishing management, health and reproduction (Collier and Beede, 1985; Collier et al, 2005)
Among cells markedly affected by exposure to thermal stress, adipocytes are of particular interest since they fulfill the dual role of energy storage and thermal insulation (Bernabucci et al, 2009)
We investigated the effect of Angelica gigas Nakai (AGN) root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible molecular mechanism of action
Summary
Heat stress imperils animal production efficiency through reduction of feed intake and diminishing management, health and reproduction (Collier and Beede, 1985; Collier et al, 2005). Aside from the negative impacts on production parameters, heat load increases health care cost that can eventually lead to an animal succumbing to severe or chronic thermal stress Cells from all organisms respond to physiologically relevant variations in temperature by rapidly increasing the expression and synthesis of a selected group of proteins, the heat shock. In a study conducted by Ezure and Amano (2009) about the effects of heat stimulation on adipogenesis, they found out that heat stimulation reduces adipogenesis by decreasing the expression of adipogenesisrelated transcriptional factors
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