性激素对3T3-L1脂肪细胞Visfatin表达的影响

Regulation of Sex Steroid Hormone on Visfatin Expression in Adipocytes and Preadipocytes

目的观察雌二醇、睾酮和孕酮对3T3-L1脂肪细胞Visfatin mRNA和蛋白表达的影响。方法 10-8mol/L～10-6mol/L雌二醇、睾酮或孕酮作用于3T3-L1成熟脂肪细胞和前脂肪细胞,孵育过夜后收集细胞,分别采用RT-PCR和Western blot检测Visfatin mRNA和蛋白的表达情况。结果在3T3-L1成熟脂肪细胞,与对照组相比,雌二醇和睾酮分别使Visfatin mRNA表达增加24%(1.74±0.31比1.40±0.18,P<0.05)和28%(1.65±0.90比1.29±0.69,P<0.05);而孕酮不影响成熟脂肪细胞Visfatin mRNA表达。雌二醇轻度增加成熟脂肪细胞Visfa-tin蛋白表达,但无统计学差异;10-6mol/L睾酮使成熟脂肪细胞Visfatin蛋白表达增加134%(0.61±0.40比0.26±0.05,P<0.05)。与雌二醇和睾酮不同,孕酮使成熟脂肪细胞Visfatin蛋白表达下调32%(0.19±0.02比0.28±0.02,P<0.05)。在前脂肪细胞,与对照组相比,10-7mol/L和10-6mol/L雌二醇使Visfatin mRNA表达增加70%(1.04±0.38比0.61±0.16,P<0.01)和123%(1.36±0.41比0.61±0.16,P<0.01);睾酮使Visfatin mRNA表达增加76%(1.02±0.24比0.58±0.36,P<0.05);孕酮使前脂肪细胞Visfatin mRNA表达增加2.6倍(1.53±1.01比0.42±0.14,P<0.05)。结论性激素通过促进或抑制脂肪细胞Visfatin基因或蛋白的表达,参与调节上述激素引起的脂肪细胞胰岛素抵抗的病理生理过程。

Aim To examine the effect of estradiol, testosterone and progesterone on Visfatin gene and protein expression in cultured 3T3-L1 adipocytes and preadipocytes. Methods 0 mmol/L （sex hormones-free DMEM/F12） , 10-8 moL/L, 10-7 mol/L and 10-6 mol/L estradiol or testosterone or progesterone were added to cultured 3T3-L1 adipo- cytes or preadipocytes overnight. Total RNA and proteins were extracted. Then the expression of Visfatin mRNA and protein was measured by RT-PCR and Western blot, respectively. Results Overnight incubation with estradiol in- creased Visfatin mRNA expression in both adipocytes （24% to 21% , P 〈 0. 05 ） at the concentration of 10-s mol/L to 10-7 mol/L and preadipocytes （70% to 123% , P 〈 0.01 ） at 10-7 mol/L to 10 -6 mol/L, respectively. Visfatin mRNA expression increased by 28% （ P 〈 0. 05 ） at 10 -s mol/L testosterone in adipocytes, 76% （P 〈 0. 05） at 10 -7 mol/L testos- terone in preadipocytes. Progesterone did not affect Visfatin mRNA expression on adipoeytes, but had a marked increase in Visfatin expression in preadipocytes by 2. 6-fold （P 〈 0. 05 ）. In 3T3-L1 adipocytes, estradiol had small but insignifi- cant effects on Visfatin protein expression, whereas testosterone had a much greater effect, increasing Visfatin protein by 134% at 10-6 mol/L （P 〈0. 05）. However, progesterone significantly decreased Visfatin protein by 32% at 10-6 mol/L （P 〈0. 05 ）. Conclusions These data suggest that Visfatin may play a physiological role in the insulin resistance caused by estradiol, testosterone or progesterone in 3T3-L1 adipocytes and preadipocytes.