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Effects of Resvega on Inflammasome Activation in Conjunction with Dysfunctional Intracellular Clearance in Retinal Pigment Epithelial (RPE) Cells
Antioxidants ( IF 6.0 ) Pub Date : 2021-01-07 , DOI: 10.3390/antiox10010067
Niina Bhattarai , Niina Piippo , Sofia Ranta-aho , Yashavanthi Mysore , Kai Kaarniranta , Anu Kauppinen

Age-related macular degeneration (AMD) is an eye disease in which retinal pigment epithelium (RPE) cells play a crucial role in maintaining retinal homeostasis and photoreceptors’ functionality. During disease progression, there is increased inflammation with nucleotide-binding domain, leucine-rich repeat, and Pyrin domain 3 (NLRP3) inflammasome activation, oxidative stress, and impaired autophagy in RPE cells. Previously, we have shown that the dietary supplement Resvega reduces reactive oxygen species (ROS) production and induces autophagy in RPE cells. Here, we investigated the ability of Resvega to prevent NLRP3 inflammasome activation with impaired protein clearance in human RPE cells. Cell viability was measured using the lactate dehydrogenase (LDH) and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Enzyme-linked immunosorbent assays (ELISA) were utilized to determine the secretion of cytokines, NLRP3, and vascular endothelial growth factor (VEGF). Caspase-1 activity was measured with a fluorescent labeled inhibitor of caspase-1 (FLICA; FAM-YVAD-FMK) and detected microscopically. Resvega improved the cell membrane integrity, which was evident as reduced LDH leakage from cells. In addition, the caspase-1 activity and NLRP3 release were reduced, as was the secretion of two inflammatory cytokines, interleukin (IL)-1β and IL-8, in IL-1α-primed ARPE-19 cells. According to our results, Resvega can potentially reduce NLRP3 inflammasome-mediated inflammation in RPE cells with impaired protein clearance.

中文翻译:

Resvega对视网膜色素上皮(RPE)细胞功能异常的细胞内清除与炎性体活化结合的影响

年龄相关性黄斑变性(AMD)是一种眼病,其中视网膜色素上皮(RPE)细胞在维持视网膜稳态和感光器功能方面起着至关重要的作用。在疾病进展过程中,RPE细胞中的核苷酸结合结构域,富含亮氨酸的重复序列和Pyrin结构域3(NLRP3)炎性小体激活,氧化应激和自噬受损,导致炎症增加。以前,我们已经表明膳食补充剂Resvega减少了活性氧(ROS)的产生并诱导RPE细胞自噬。在这里,我们研究了Resvega预防人RPE细胞中蛋白质清除受损的NLRP3炎性体活化的能力。使用乳酸脱氢酶(LDH)和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)测定来测量细胞活力。酶联免疫吸附测定(ELISA)用于确定细胞因子,NLRP3和血管内皮生长因子(VEGF)的分泌。用荧光标记的caspase-1抑制剂(FLICA; FAM-YVAD-FMK)测量caspase-1活性,并在显微镜下进行检测。Resvega改善了细胞膜的完整性,这可以从细胞中减少LDH泄漏中看出。另外,在IL-1α引发的ARPE-19细胞中,胱天蛋白酶-1活性和NLRP3释放降低,以及两种炎性细胞因子白介素(IL)-1β和IL-8的分泌也降低。根据我们的研究结果,Resvega可以潜在地减轻RPE细胞中NLRP3炎性体介导的炎症,并降低蛋白清除率。用荧光标记的caspase-1抑制剂(FLICA; FAM-YVAD-FMK)测量caspase-1活性,并在显微镜下进行检测。Resvega改善了细胞膜的完整性,这可通过减少细胞中LDH的泄漏来体现。另外,在IL-1α引发的ARPE-19细胞中,胱天蛋白酶-1活性和NLRP3释放降低,以及两种炎性细胞因子白介素(IL)-1β和IL-8的分泌也降低。根据我们的研究结果,Resvega可以潜在地减轻RPE细胞中NLRP3炎性体介导的炎症,并降低蛋白清除率。用荧光标记的caspase-1抑制剂(FLICA; FAM-YVAD-FMK)测量caspase-1活性,并在显微镜下进行检测。Resvega改善了细胞膜的完整性,这可通过减少细胞中LDH的泄漏来体现。另外,在IL-1α引发的ARPE-19细胞中,胱天蛋白酶-1活性和NLRP3释放降低,以及两种炎性细胞因子白介素(IL)-1β和IL-8的分泌也降低。根据我们的研究结果,Resvega可以潜在地减轻RPE细胞中NLRP3炎性体介导的炎症,并降低蛋白清除率。以及在IL-1α引发的ARPE-19细胞中两种炎性细胞因子白介素(IL)-1β和IL-8的分泌。根据我们的研究结果,Resvega可以潜在地减轻RPE细胞中NLRP3炎性体介导的炎症,并降低蛋白清除率。以及在IL-1α引发的ARPE-19细胞中两种炎性细胞因子白介素(IL)-1β和IL-8的分泌。根据我们的研究结果,Resvega可以潜在地减轻RPE细胞中NLRP3炎性体介导的炎症,并降低蛋白清除率。
更新日期:2021-01-07
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