352 微神经胶质细胞行为和 Iba-1 表达:评估血管性痴呆和长期 COVID 对认知的影响

Grant Talkington, Saifudeen Ismael, T. Gressett, G. Bix
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摘要

目的/目标:本研究旨在探讨小胶质细胞行为在与血管性痴呆(VaD)和长COVID相关的认知障碍中的作用。我们将利用免疫组化(IHC)和定量 PCR(qPCR)技术,评估 VaD 和 SARS-CoV-2 感染者体内小胶质细胞活化标志物 Iba-1 的表达情况。方法/研究对象:在 48 只雌性 C57BL/6 小鼠中,有 24 只接受了双侧颈动脉狭窄(BCAS)手术干预,以实验性诱导血管性痴呆。2 周后,12 只 BCAS 小鼠和 12 只非 BCAS 小鼠分别感染了 1E4 PFU 的小鼠适应型 10(MA10)SARS-CoV-2 株。感染后 2 周,术后 4 周,所有动物均被安乐死,并对组织进行 cDNA 和组织学处理。使用免疫荧光和 RT-qPCR 通过 Iba-1 对小胶质细胞进行量化,通过 claudin-5 以及 occludin、GFAP 和 integrin a5 对 BBB 的完整性进行量化。结果/预期结果:我们预计会在血管性痴呆(VaD)患者和长COVID患者中观察到不同的小胶质细胞行为模式。通过免疫组化 (IHC),我们预计会看到 Iba-1 表达增加,这表明小胶质细胞活化。定量 PCR (qPCR) 可能会证实这些发现,显示 Iba-1 mRNA 水平升高。最后,我们预计数据将揭示小胶质细胞与血脑屏障(BBB)之间的相互作用。这些相互作用可让我们深入了解小胶质细胞的行为如何影响 BBB 的完整性,进而影响 VaD 和长 COVID 患者的认知功能。讨论/意义:本研究旨在阐明小胶质细胞在与血管性痴呆和长程COVID相关的认知功能下降中的作用。通过根据小胶质细胞的激活情况对患者进行分类,我们可以更好地对症下药。研究结果可能会开发出针对这些疾病认知障碍的靶向疗法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
352 Microglial Behavior and Iba-1 Expression: Evaluating the Cognitive Impact of Vascular Dementia and Long COVID
OBJECTIVES/GOALS: The study aims to explore the role of microglial behavior in cognitive impairment associated with vascular dementia (VaD) and long COVID. Using immunohistochemistry (IHC) and quantitative PCR (qPCR), we will assess the expression of Iba-1, a microglial activation marker, in subjects with VaD and SARS-CoV-2 infection. METHODS/STUDY POPULATION: Out of 48 female C57BL/6 mice, 24 had surgical intervention in the form of bilateral carotid artery stenosis (BCAS) for experimental induction of vascular dementia. After 2 weeks, 12 BCAS and 12 non-BCAS were infected with 1E4 PFU of mouse-adapted 10 (MA10) strain of SARS-CoV-2. 2 weeks post-infection, 4 weeks post-operatively, all animals were euthanized and tissues were processed for cDNA and histology. Immunofluorescence and RT-qPCR used to quantify microglia via Iba-1, BBB integrity via claudin-5 as well as occludin, GFAP, and integrin a5. RESULTS/ANTICIPATED RESULTS: We anticipate observing distinct patterns of microglial behavior in subjects with vascular dementia (VaD) and those with long COVID. Through immunohistochemistry (IHC), we expect to see increased Iba-1 expression, indicative of microglial activation. Quantitative PCR (qPCR) will likely corroborate these findings, showing elevated levels of Iba-1 mRNA. Lastly, we anticipate that the data will reveal interactions between microglia and the blood-brain barrier (BBB). These interactions could provide insights into how microglial behavior influences BBB integrity and, consequently, cognitive function in VaD and long COVID. DISCUSSION/SIGNIFICANCE: This study aims to clarify the role of microglia in cognitive decline linked to vascular dementia and long COVID. By categorizing patients based on microglial activation, we can better tailor treatments. The findings could lead to targeted therapies that address cognitive impairment in these conditions.
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