Programa de Pós-Graduação em Ciências Biológicas-Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul - UFRGS, Porto Alegre, Rio Grande do Sul, Brasil.
Department of Clinical Nutrition, Institute of Public Health and Clinical Nutrition, University of Eastern Finland, Kuopio, Finland.
Cell Biol Int. 2022 Nov;46(11):1787-1800. doi: 10.1002/cbin.11876. Epub 2022 Aug 16.
Caveolin-1 (Cav-1) is an integral membrane protein present in all organelles, responsible for regulating and integrating multiple signals as a platform. Mitochondria are extremely adaptable to external cues in chronic liver diseases, and expression of Cav-1 may affect mitochondrial flexibility in hepatic stellate cells (HSCs) activation. We previously demonstrated that exogenous expression of Cav-1 was sufficient to increase some classical markers of activation in HSCs. Here, we aimed to evaluate the influence of exogenous expression and knockdown of Cav-1 on regulating the mitochondrial plasticity, metabolism, endoplasmic reticulum (ER)-mitochondria distance, and lysosomal activity in HSCs. To characterize the mitochondrial, lysosomal morphology, and ER-mitochondria distance, we perform transmission electron microscope analysis. We accessed mitochondria and lysosomal networks and functions through a confocal microscope and flow cytometry. The expression of mitochondrial machinery fusion/fission genes was examined by real-time polymerase chain reaction. Total and mitochondrial cholesterol content was measured using Amplex Red. To define energy metabolism, we used the Oroboros system in the cells. We report that GRX cells with exogenous expression or knockdown of Cav-1 changed mitochondrial morphometric parameters, OXPHOS metabolism, ER-mitochondria distance, lysosomal activity, and may change the activation state of HSC. This study highlights that Cav-1 may modulate mitochondrial function and structural reorganization in HSC activation, being a potential candidate marker for chronic liver diseases and a molecular target for therapeutic intervention.
窖蛋白-1(Cav-1)是一种存在于所有细胞器中的完整膜蛋白,作为平台负责调节和整合多种信号。线粒体在慢性肝脏疾病中对外部刺激极其适应,而 Cav-1 的表达可能会影响肝星状细胞(HSCs)激活中的线粒体灵活性。我们之前的研究表明,外源性表达 Cav-1足以增加 HSCs 中一些经典的激活标志物。在这里,我们旨在评估外源性表达和敲低 Cav-1 对调节 HSCs 中线粒体可塑性、代谢、内质网(ER)-线粒体距离和溶酶体活性的影响。为了表征线粒体、溶酶体形态和 ER-线粒体距离,我们进行了透射电子显微镜分析。我们通过共聚焦显微镜和流式细胞术来评估线粒体和溶酶体网络和功能。通过实时聚合酶链反应检查线粒体机器融合/裂变基因的表达。使用 Amplex Red 测量总胆固醇和线粒体胆固醇含量。为了定义能量代谢,我们在细胞中使用了 Oroboros 系统。我们报告说,外源性表达或敲低 Cav-1 的 GRX 细胞改变了线粒体形态计量参数、OXPHOS 代谢、ER-线粒体距离、溶酶体活性,并可能改变 HSC 的激活状态。这项研究强调,Cav-1 可能调节 HSC 激活中线粒体功能和结构重排,是慢性肝脏疾病的潜在候选标志物和治疗干预的分子靶点。