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Abstract
Purpose:
The objective of this study was to investigate the effects of (6)-gingerol, ginger components proliferation and adipocyte differentiation from early to lately steps.
Methods:
3T3-L1 preadipocytes were cultured. Differentiation of confluent cells was induced with dexamethasone, isobutylxanthin and insulin for 2 day and cells were cultured by medium with insulin in presence of various concentrations 0, 25, 50, 100 (μmol/L) of (6)-gingerol for 4 day. Cell viability was measured using the EZ Cytox assay kit. In addition, we examined the expression of mRNA levels associated with each adipocyte differentiation step by real time reverse transcription polymerase chain reaction.
Results:
(6)-Gingerol inhibited adipocyte proliferation in a dose and time dependent manner. Expression of C/EBPβ, associated with early differentiation step remained unchaged. However, intermmediate, late differentiation step and adipocytokines were effectively changed in dose-dependently manner in cell groups treated with (6)-gingerol.
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Fig. 1.
Effect of (6)-gingerol on cell proliferation in 3T3-L1 preadipocyte. 3T3-L1 cells were plated at a density of 0.5×104 cell/mL in a 96 well plate with DMEM supplemented with 10% BCS for 2 day, the monolayers were differentiation induction with DMEM supplemented with 10% FBS, 10 μg/ml insulin, 1 μM/ml Dex, 0.5 mM/ml IBMX for 2 day. After differentiation induction, the monolayer were incubated in postdifferentiation medium with 0, 25, 50, 100 μM (6)-gingerol. Viable cell numbers were estimated by the EZ-Cytox cell viability assay. Each bar represents the mean ± SE Comparison among different concentration of α-lipoic acid that yielded significant differences (p < 0.05) are indicated by different letters above each bar.
Fig. 2.
Effects of (6)-gingerol on mRNA expression of β-actin and C/EBPβ expression in 3T3-L1 adipocyte. Total RNA was isolated using TRI-reagent and cDNA was synthesized using 3 μg of total RNA with SuperScrit II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne software v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α=0.05 as determined by Duncan's multiple range test.
Fig. 3.
Effects of (6)-gingerol on mRNA expression of PPARγ and C/ EBPα expression in 3T3-L1 adipocyte. Total RNA was isolated using TRI-reagent and cDNA was synthesized using 3 μg of total RNA with SuperScrit II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne software v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α=0.05 as determined by Duncan's multiple range test.
Fig. 4.
Effects of (6)-gingerol on mRNA expression of FABP4 and AP2 expression in 3T3-L1 adipocyte. Total RNA was isolated using TRI-reagent and cDNA was synthesized using 3 μg of total RNA with SuperScrit II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne software v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α=0.05 as determined by Duncan's multiple range test.
Fig. 5.
Effects of (6)-gingerol on mRNA expression of leptin and adiponectin expression in 3T3-L1 adipocyte. Total RNA was isolated using TRI-reagent and cDNA was synthesized using 3 μg of total RNA with SuperScrit II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne software v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α=0.05 as determined by Duncan's multiple range test.
Table 1.
Sequence and RT-PCR program of transcription factor
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