Tanhapour Maryam, Nourbakhsh Mitra, Panahi Ghodratollah, Golestani Abolfazl
Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Department of Clinical Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
J Diabetes Metab Disord. 2024 Mar 7;23(1):1081-1091. doi: 10.1007/s40200-024-01389-4. eCollection 2024 Jun.
Considering inhibition of pre-adipocyte cells differentiation in adipose tissue fibrosis, we aimed to explore whether Sirt1 and Hif-1α in pre-adipocytes have a significant effect on fibrotic gene expression.
3T3-L1 pre-adipocytes were transfected with SIRT1-specific siRNA, confirmed by real-time polymerase chain reaction (RT-PCR) and western blotting. Additionally, cells were treated with varying concentrations of resveratrol and sirtinol as the activator and inhibitor of Sirt1, respectively. Involvement of Hif-1α was evaluated by treatment with echinomycin. Subsequently, we assessed the gene and protein expressions related to fibrosis in the extracellular matrix of adipose tissue, including collagen VI (), lysyl oxidase (), matrix metalloproteinase-2 (), , and osteopontin () in pre-adipocytes through RT-PCR and western blot.
The current study demonstrated that knockdown and reduced enzyme activity significantly increased the expression of , , , , and genes in the treated 3T3-L1 cells compared to the control group. Interestingly, resveratrol significantly decreased the gene expression related to the fibrosis pathway. Inhibition of by echinomycin led to a significant reduction in , , and gene expression in the treated group compared to the control.
This study highlights that down-regulation of might be a predisposing factor in the emergence of adipose tissue fibrosis by enhancing the expression of extracellular matrix (ECM) components. Activation of , similar to suppressing of in pre-adipocytes may be a beneficial approach for attenuating fibrotic gene expression.
The online version contains supplementary material available at 10.1007/s40200-024-01389-4.
鉴于脂肪组织纤维化过程中前脂肪细胞分化受到抑制，我们旨在探讨前脂肪细胞中的Sirt1和Hif-1α对纤维化基因表达是否有显著影响。
用SIRT1特异性小干扰RNA转染3T3-L1前脂肪细胞，通过实时聚合酶链反应（RT-PCR）和蛋白质免疫印迹法进行验证。此外，分别用不同浓度的白藜芦醇和sirtinol处理细胞，白藜芦醇和sirtinol分别作为Sirt1的激活剂和抑制剂。用放线菌素处理以评估Hif-1α的参与情况。随后，我们通过RT-PCR和蛋白质免疫印迹法评估前脂肪细胞中脂肪组织细胞外基质中与纤维化相关的基因和蛋白质表达，包括Ⅵ型胶原、赖氨酰氧化酶、基质金属蛋白酶-2、以及骨桥蛋白。
当前研究表明，与对照组相比，基因敲低和酶活性降低显著增加了处理后的3T3-L1细胞中、、、、和基因的表达。有趣的是，白藜芦醇显著降低了与纤维化途径相关的基因表达。与对照组相比，放线菌素对的抑制导致处理组中、和基因表达显著降低。
本研究强调，通过增强细胞外基质（ECM）成分的表达，下调可能是脂肪组织纤维化出现的一个易感因素。在原代脂肪细胞中激活，类似于抑制，可能是减弱纤维化基因表达的一种有益方法。
网络版包含可在10.1007/s40200-024-01389-4获取的补充材料。
