烧伤导致紧急造血过程中的髓系引物。

IF 2.7 3区 医学 Q2 CRITICAL CARE MEDICINE
SHOCK Pub Date : 2024-08-13 DOI:10.1097/SHK.0000000000002458
Ryan M Johnson, Kevin E Galicia, Huashan Wang, Richard Gonzalez, Mashkoor Choudhry, John Kubasiak
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引用次数: 0

摘要

简介造血以分化模式进行,从造血干细胞(HSC)开始,最终形成红系、髓系和淋巴系。造血干系承诺的病理性改变可导致白细胞生成不足或细胞系功能失调。烧伤后紧急造血的驱动因素尚不明确。烧伤引起的髓系占优势与感染有关,而感染会恶化预后。本研究旨在进一步分析小鼠烧伤后骨髓造血干细胞的情况:方法:C57BL/6小鼠背侧总体表面积(TBSA)约12%的烫伤部位接受烧伤或假性损伤,随后在伤后1、2、3、7和10天处死。通过流式细胞术分析后肢骨髓(BM)中的造血干细胞群,并使用 FlowJo 软件(10.6 版)进行分析。采用多重非配对 t 检验和线性混合效应回归分析事件计数和频率。针对 PU.1、GATA-1 和 GATA-3 对分离的系阴性 BM 细胞 RNA 进行 RT-PCR,随后用 QuantStudio 3 软件进行分析。使用 GraphPad 软件(Prism)进行统计分析和表示:流式细胞术显示,烧伤小鼠的长期造血干细胞比例在伤后 3 天明显升高(p < .05),短期造血干细胞比例在伤后 2、3 和 10 天明显升高(均 p < .05)。与假对照组相比,烧伤组群中多能祖细胞(MPP)2 和 3 亚群的比例持续上升,但并不显著。普通髓系祖细胞(CMP)的比例在第 3 天和第 10 天显著升高(均 p <.01),而粒细胞-巨噬细胞祖细胞(GMP)的比例在第 1 天、第 2 天和第 10 天升高(p <.05、p <.01、p <.01)。虽然烧伤组群中巨核细胞-红细胞祖细胞(MEP)的比例持续降低,但未达到显著性水平。mRNA 分析结果显示,PU.1 在第 1 天出现下调(p = 0.0002),在第 7 天出现上调(p < 0.01)。GATA-1 在第 7 天出现下调(p < 0.05),GATA3 在第 3 天和第 7 天出现下调(p < 0.05):讨论:全厚烧伤导致紧急造血,从伤后早期开始,长期造血干细胞和短期造血干细胞/MPP1亚群按比例增加。随后的血系形成以髓系为主,并转向髓系祖细胞,mRNA 分析证实了这一发现,PU.1 上调,GATA-1 和 GATA-3 下调。要了解烧伤诱导的紧急造血是如何通过病理血系选择导致感染的,还需要进一步的研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Burn Injury Results in Myeloid Priming during Emergency Hematopoiesis.

Introduction: Hematopoiesis proceeds in a tiered pattern of differentiation, beginning with hematopoietic stem cells (HSC) and culminating in erythroid, myeloid, and lymphoid lineages. Pathologically altered lineage commitment can result in inadequate leukocyte production or dysfunctional cell lines. Drivers of emergency hematopoiesis after burn injury are inadequately defined. Burn injury induces a myeloid predominance associated with infection that worsens outcomes. This study aims to further profile bone marrow HSCs following burn injury in a murine model.

Methods: C57BL/6 mice received burn or sham injury with ~12% total body surface area (TBSA) scald burn on the dorsal surface with subsequent sacrifice at 1, 2, 3, 7 and 10 days post-injury. Bone marrow (BM) from hindlimbs were analyzed for HSC populations via flow cytometry and analyzed using FlowJo Software (version 10.6). Event counts and frequencies were analyzed with multiple unpaired t-tests and linear mixed-effect regression. RT-PCR performed on isolated lineage negative BM cell RNA targeted PU.1, GATA-1, and GATA-3 with subsequent analysis conducted with QuantStudio 3 software. Statistical analysis and representation were performed on GraphPad software (Prism).

Results: Flow cytometry revealed significantly elevated proportions of Long-Term HSCs at 3 days post-injury (p < .05) and Short-Term HSCs at days 2, 3, and 10 (all p < .05) in burn-injured mice. There was a sustained, but not significant, increase in proportions in the multi-potent progenitor (MPP) 2 and 3 subpopulations in the burn cohort compared to sham controls. The common myeloid progenitor (CMP) proportion was significantly higher on days 3 and 10 (both p < .01), while the granulocyte-macrophage progenitor (GMP) proportion increased on days 1, 2, and 10 (p < .05, p < .01, p < .01). Although the megakaryocyte-erythrocyte progenitor (MEP) proportion appeared consistently lower in the burn cohort, this did not reach significance. mRNA analysis resulted in a downregulation of PU.1 on day 1 (p = 0.0002) with an upregulation by day 7 (p < 0.01). GATA-1 downregulation occurred by day 7 (p < 0.05), and GATA3 showed downregulation on days 3 and 7 (p < 0.05).

Discussion: Full-thickness burn results in an emergency hematopoiesis via proportional increase of Long Term-HSC and Short Term-HSC/MPP1 subpopulations beginning in the early post-injury period. Subsequent lineage commitment displays a myeloid predominance with a shift toward myeloid progenitors with mRNA analysis corroborating this finding with associated upregulation of PU.1 and downregulation of GATA-1 and GATA-3. Further studies are needed to understand how burn-induced emergency hematopoiesis may predispose to infection by pathologic lineage selection.

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来源期刊
SHOCK
SHOCK 医学-外科
CiteScore
6.20
自引率
3.20%
发文量
199
审稿时长
1 months
期刊介绍: SHOCK®: Injury, Inflammation, and Sepsis: Laboratory and Clinical Approaches includes studies of novel therapeutic approaches, such as immunomodulation, gene therapy, nutrition, and others. The mission of the Journal is to foster and promote multidisciplinary studies, both experimental and clinical in nature, that critically examine the etiology, mechanisms and novel therapeutics of shock-related pathophysiological conditions. Its purpose is to excel as a vehicle for timely publication in the areas of basic and clinical studies of shock, trauma, sepsis, inflammation, ischemia, and related pathobiological states, with particular emphasis on the biologic mechanisms that determine the response to such injury. Making such information available will ultimately facilitate improved care of the traumatized or septic individual.
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