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白藜芦醇对猪原代前脂肪细胞凋亡的影响及潜在机制

[Effect and underlying mechanism of resveratol on porcine primary preadipocyte apoptosis].

作者信息

Zhang Zhao, Yang Yang, Pang Weijun, Sun Chao, Yang Gongshe

机构信息

Laboratory of Animal Fat Deposition and Muscle Development, Northwest A&F University, Yangling 712100, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2010 Aug;26(8):1042-9.

Abstract

We demonstrated the effect of resveratrol on porcine primary preadipocytes apoptosis, to study the intracellular molecular mechanism. Porcine primary preadipocyte was treated with different concentration of resveratrol (0 micromol/L, 50 micromol/L, 100 micromol/L, 200 micromol/L, 400 micromol/L). We used optical microscope and fluorescence microscope to observe morphological changes during apoptosis after Hoechst 33258 Fluorescent dyes staining; and RT-PCR and Western blotting to measure the expression of apoptosis-associated gene sirt1, caspase-3, bcl-2, bax, p53, NF-kappaB. Primary preadipocyte apoptosis was apparent, accompanied by reduced cell volume, chromatin condensation, and nuclear shrinkage. Compared to the control and low concentration group, high dose group (200 micromol/L) significantly increased the ratio of primary preadipocyte apoptosis. The expression of sirt1, caspase-3, and bax was up-regulated markedly in response to resveratrol; in contrast, apoptotic inhibitor bcl-2, p53, NF-kappaB down-regulated. We further proved fact that resveratrol can specifically promote the activity of sirt1; moreover, activated sirt1 modulates the activity of caspase-3 and bcl-2 family, involving in transcriptional regulation of p53 and NF-kappaB through antagonizing factor-induced acetylation. Taken together, our data established resveratrol as new regulator in porcine primary preadipocyte apoptosis via activating the expression of sirt1, modulating activity of apoptotic-associated factor.

摘要

我们展示了白藜芦醇对猪原代前脂肪细胞凋亡的影响,以研究其细胞内分子机制。用不同浓度的白藜芦醇(0微摩尔/升、50微摩尔/升、100微摩尔/升、200微摩尔/升、400微摩尔/升)处理猪原代前脂肪细胞。在经Hoechst 33258荧光染料染色后,我们使用光学显微镜和荧光显微镜观察凋亡过程中的形态变化;并通过RT-PCR和蛋白质印迹法检测凋亡相关基因sirt1、半胱天冬酶-3、bcl-2、bax、p53、核因子κB的表达。原代前脂肪细胞凋亡明显,伴有细胞体积减小、染色质凝聚和核固缩。与对照组和低浓度组相比,高剂量组(200微摩尔/升)显著增加了原代前脂肪细胞凋亡率。白藜芦醇显著上调了sirt1、半胱天冬酶-3和bax的表达;相反,凋亡抑制因子bcl-2、p53、核因子κB表达下调。我们进一步证实了白藜芦醇能特异性促进sirt1的活性;此外,激活的sirt1通过拮抗因子诱导的乙酰化作用调节半胱天冬酶-3和bcl-2家族的活性,参与p53和核因子κB的转录调控。综上所述,我们的数据表明白藜芦醇通过激活sirt1的表达、调节凋亡相关因子的活性,成为猪原代前脂肪细胞凋亡的新调节因子。

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