2095-LB：SARS-CoV-2 Spike 蛋白对胰岛微血管功能的影响

IF 6.2 1区医学 Q1 ENDOCRINOLOGY & METABOLISM

Diabetes Pub Date : 2024-06-21 DOI:10.2337/db24-2095-lb

CATARINA BARBOZA, LUCIANA MATEUS GONCALVES, JOANA ALMACA

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引用次数: 0

摘要

导言冠状病毒病 2019（COVID-19）是由严重急性呼吸系统综合征冠状病毒-2（SARS-CoV-2）引起的。我们假设，SARS-CoV-2 可通过损害胰腺微血管功能引发葡萄糖稳态丧失。研究方法我们用 SARS-CoV-2 Spike S1 重组蛋白（SARS-Spike；80 nM，5xEC50，1 小时）培养来自 7 名非糖尿病人类供体（来自 nPOD）的活体胰腺切片。作为对照，我们用来自 HCoV-OC43 的 Spike S1 蛋白培养切片。通过共聚焦显微镜，我们监测了这些 Spike S1 蛋白对周皮[Ca2+]i 反应和血管运动（用荧光凝集素标记的毛细血管）的急性影响。通过酶联免疫吸附试验，我们评估了 Spike S1 蛋白对人胰腺切片上清液中血管紧张素 II 和血管紧张素 1-7 内源性水平的影响。实验结果用 SARS-Spike 急性孵育可导致活体切片中人胰岛毛细血管的关闭。然后，我们在 3 mM 葡萄糖溶液中用血管紧张素 II（100 nM，持续 4 分钟）刺激切片。用 HCoV-OC43 培养的切片中的胰岛毛细血管在血管紧张素 II 的作用下收缩（直径平均缩小约 14%），而用 SARS-Spike 处理的切片中的血管对血管紧张素 II 的刺激没有进一步反应。此外，用 SARS-Spike 培养可降低血管紧张素 II 对胰岛周细胞钙水平的刺激作用。用 SARS-Spike 培养可使活体胰腺切片产生的血管紧张素 II 水平略有增加，而血管紧张素 1-7 的浓度则没有差异。结论我们的数据表明，SARS-CoV-2 对血管细胞的潜在感染可能会干扰周细胞的收缩特性并损害毛细血管反应。我们将在未来确定这些变化是否与 ACE2 受抑制和/或其在质膜上的表达减少有关。披露 C. Barboza：无。L. Mateus Goncalves：无：无。J. Almaca：无：无。资助美国国立卫生研究院（R01DK133483）

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2095-LB: Impact of SARS-CoV-2 Spike Proteins on the Islet Microvascular Function

Introduction: The coronavirus disease 2019 (COVID-19) is caused by the Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). We hypothesized that SARS-CoV-2 could trigger loss of glucose homeostasis by compromising microvascular function in the pancreas. Methods: We incubated living pancreas slices from 7 non-diabetic human donors (from nPOD) with SARS-CoV-2 Spike S1 recombinant protein (SARS-Spike; 80 nM, 5xEC50, 1h). As a control, we incubated slices with a Spike S1 protein from HCoV-OC43. By confocal microscopy, we monitored the acute effects of these Spike S1 proteins on pericyte [Ca2+]i responses and vasomotion (capillaries labeled with fluorescent lectin). Using an ELISA assay, we assessed the effect of Spike S1 proteins on the endogenous levels of angiotensin II and angiotensin 1-7 in the supernatant of human pancreas slices. Results: Acute incubation with SARS-Spike led to closure of capillaries in human islets in living slices. We then stimulated slices with angiotensin II (100 nM; for 4 min) in 3 mM glucose solution. While islet capillaries in slices incubated with HCoV-OC43 constricted upon Angiotensin II application (~14% average reduction in diameter), vessels in slices treated with SARS-Spike did not further respond to Angiotensin II stimulation. In addition, incubation with SARS-Spike decreased the stimulatory effect of Angiotensin II on islet pericyte cytosolic calcium levels. Incubation with SARS-Spike slightly increased the levels of Angiotensin II produced by living pancreas slices while there was no difference in Angiotensin1-7 concentration. Conclusion: Our data indicate that the potential infection of vascular cells by SARS-CoV-2 could interfere with pericytes’ contractile properties and compromise capillary responses. We will determine in the future whether these changes are associated with an inhibition of ACE2 and/or a reduction of its expression at the plasma membrane. Disclosure C. Barboza: None. L. Mateus Goncalves: None. J. Almaca: None. Funding National Institutes of Health (R01DK133483)

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来源期刊

Diabetes 医学-内分泌学与代谢

CiteScore

12.50

自引率

2.60%

发文量

1968

审稿时长

1 months

期刊介绍： Diabetes is a scientific journal that publishes original research exploring the physiological and pathophysiological aspects of diabetes mellitus. We encourage submissions of manuscripts pertaining to laboratory, animal, or human research, covering a wide range of topics. Our primary focus is on investigative reports investigating various aspects such as the development and progression of diabetes, along with its associated complications. We also welcome studies delving into normal and pathological pancreatic islet function and intermediary metabolism, as well as exploring the mechanisms of drug and hormone action from a pharmacological perspective. Additionally, we encourage submissions that delve into the biochemical and molecular aspects of both normal and abnormal biological processes. However, it is important to note that we do not publish studies relating to diabetes education or the application of accepted therapeutic and diagnostic approaches to patients with diabetes mellitus. Our aim is to provide a platform for research that contributes to advancing our understanding of the underlying mechanisms and processes of diabetes.