Bałaban Jaśmina, Wierzbicki Mateusz, Zielińska-Górska Marlena, Sosnowska Malwina, Daniluk Karolina, Jaworski Sławomir, Koczoń Piotr, Cysewski Dominik, Chwalibog André, Sawosz Ewa
Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland.
Department of Chemistry, Institute of Food Sciences, Warsaw University of Life Sciences, Warsaw, Poland.
Nanotechnol Sci Appl. 2023 Jan 19;16:1-18. doi: 10.2147/NSA.S391761. eCollection 2023.
AIM: The experiments aimed to document the presence of the ACE2 receptor on human muscle cells and the effects of the interaction of these cells with the spike protein of the SARS-CoV-2 virus in terms of induction of pro-inflammatory proteins, as well as to assess the possibility of reducing the pool of these proteins with the use of graphene oxide (GO) flakes. METHODS: Human Skeletal Myoblast (HSkM), purchased from Gibco were maintained in standard condition according to the manufacturer's instruction. The cells were divided into 4 groups; 1. C-control, 2. S-with addition of spike protein, 3. GO-with the addition of graphene oxide, 4. GO-S-with addition of GO followed by the addition of S protein. Protein S (PX-COV-P049) was purchased from ProteoGenix (France). GO was obtained from Advanced Graphene Products (Zielona Gora, Poland). The influence of all the factors on the morphology of cells was investigated using light and confocal microscopy. ACE2 protein expression on muscle cells was visualized and 40 pro-inflammatory cytokines were investigated using the membrane antibody array method. The protein profile of the lysate of cells from individual groups was also analyzed by mass spectrometry. CONCLUSION: The experiments confirmed the presence of the ACE2 receptor in human skeletal muscle cells. It has also been documented that the SARS-CoV-2 virus spike protein influences the activation of selected pro-inflammatory proteins that promote cytokine storm and oxidative stress in muscle cells. The use of low levels of graphene oxide does not adversely affect muscle cells, reducing the levels of most proteins, including pro-inflammatory proteins. It can be assumed that GO may support anti-inflammatory therapy in muscles by scavenging proteins that activate cytokine storm.
目的:这些实验旨在证明人肌肉细胞上存在血管紧张素转换酶2(ACE2)受体,以及这些细胞与严重急性呼吸综合征冠状病毒2(SARS-CoV-2)病毒刺突蛋白相互作用在诱导促炎蛋白方面的影响,并评估使用氧化石墨烯(GO)薄片减少这些蛋白总量的可能性。 方法:从Gibco购买的人骨骼肌成肌细胞(HSkM)按照制造商的说明在标准条件下培养。细胞分为4组:1. C-对照组;2. S-添加刺突蛋白组;3. GO-添加氧化石墨烯组;4. GO-S-先添加GO后添加S蛋白组。蛋白S(PX-COV-P049)购自法国的ProteoGenix公司。GO购自波兰齐洛纳古拉的Advanced Graphene Products公司。使用光学显微镜和共聚焦显微镜研究所有因素对细胞形态的影响。通过膜抗体阵列法观察肌肉细胞上ACE2蛋白的表达,并研究40种促炎细胞因子。还通过质谱分析了各个组细胞裂解物的蛋白质谱。 结论:实验证实了人骨骼肌细胞中存在ACE2受体。还证明了SARS-CoV-2病毒刺突蛋白会影响特定促炎蛋白的激活,这些蛋白会促进肌肉细胞中的细胞因子风暴和氧化应激。使用低水平的氧化石墨烯不会对肌肉细胞产生不利影响,反而会降低大多数蛋白的水平,包括促炎蛋白。可以推测,GO可能通过清除激活细胞因子风暴的蛋白来支持肌肉的抗炎治疗。
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