损伤后阻断 RAGE 表达可减少急性肺损伤小鼠模型中的炎症反应。

Blocking RAGE expression after injury reduces inflammation in mouse model of acute lung injury.

机构信息

Departments of Medicine, Anesthesiology, and Pathology, Columbia University, 622 West 168th St, PH 10-203, New York, NY, 10032, USA.

出版信息

Respir Res. 2023 Jan 20;24(1):21. doi: 10.1186/s12931-023-02324-6.

DOI:10.1186/s12931-023-02324-6

PMID:36670409

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9852798/

Abstract

BACKGROUND

Receptor for Advanced Glycated Endproducts (RAGE) plays a major role in the inflammatory response to infectious and toxin induced acute lung injury. We tested the hypothesis that a RAGE blocking antibody when administered after the onset of injury can reduce lung inflammation compared to control antibody.

METHODS

Male and female C57BL/6 (WT) mice were used. Forty-six received lipopolysaccharide (LPS) and 26 PBS by nasal instillation on day one, repeated on day three. On day 2, 36 mice receiving LPS were divided into two groups of 18, one treated with 200 μg of non-immune isotype control IgG and the second group treated with 200 μg of anti-RAGE Ab, each dose divided between IV and IP. Ten of the 46 were not treated. On day 4, before euthanasia, mice were injected with fluorescein isothiocyanate (FITC) labelled albumen. BALF and serum samples were collected as well as lung tissue for immunohistochemistry (IHC). BALF was analyzed for cell (leukocyte) counts, for FITC BALF/serum ratios indicating pulmonary vascular leak, and for cytokines/chemokines using bead based multiplex assays. Quantitative IHC was performed for MPO and RAGE.

RESULTS

Ten LPS mice showed minimal inflammation by all measures indicating poor delivery of LPS and were excluded from analysis leaving n = 11 in the LPS + IgG group and n = 12 in the LPS + anti-RAGE group. BALF cell counts were low in the PBS administered mice (4.9 ± 2.1 × 10/ml) and high in the LPS injured untreated mice (109 ± 34) and in the LPS + IgG mice (91 ± 54) while in comparison, LPS + anti-RAGE ab mice counts were significantly lower (51.3 ± 18 vs. LPS + IgG, P = 0.03). The BALF/serum FITC ratios were lower for the LPS + anti-RAGE mice than for the LPS + IgG mice indicating less capillary leakiness. Quantitative IHC RAGE staining was lower in the LPS + anti-RAGE ab mice than in the LPS + IgG treated mice (P = 0.02).

CONCLUSIONS

These results describe a four-day LPS protocol to sustain lung injury and allow for treatment and suggests that treatment aimed at blocking RAGE when given after onset of injury can reduce lung inflammation.

摘要

背景

晚期糖基化终产物受体（RAGE）在感染和毒素诱导的急性肺损伤的炎症反应中起主要作用。我们检验了这样一种假设，即在损伤发生后给予 RAGE 阻断抗体可以减少与对照抗体相比的肺炎症。

方法

使用雄性和雌性 C57BL/6（WT）小鼠。46 只接受脂多糖（LPS）和 26 只 PBS 通过鼻腔滴注在第 1 天，第 3 天重复。在第 2 天，36 只接受 LPS 的小鼠分为两组，每组 18 只，一组用 200μg 非免疫同种型对照 IgG 治疗，另一组用 200μg 抗 RAGE Ab 治疗，每个剂量分为 IV 和 IP。46 只中有 10 只未治疗。在第 4 天，安乐死前，用荧光素异硫氰酸酯（FITC）标记白蛋白注射小鼠。收集 BALF 和血清样本以及肺组织进行免疫组织化学（IHC）。通过基于珠子的多重分析检测 BALF 中的细胞（白细胞）计数、指示肺血管渗漏的 FITC BALF/血清比以及细胞因子/趋化因子。进行定量 IHC 以检测 MPO 和 RAGE。

结果

10 只 LPS 小鼠的所有指标均显示出最小的炎症，表明 LPS 的传递不良，因此未纳入分析，留下 LPS+IgG 组 n=11 只，LPS+抗 RAGE 组 n=12 只。PBS 给药小鼠的 BALF 细胞计数较低（4.9±2.1×10/ml），未治疗 LPS 损伤的小鼠和 LPS+IgG 小鼠的 BALF 细胞计数较高（109±34），而 LPS+抗 RAGE ab 小鼠的计数明显较低（51.3±18 vs LPS+IgG，P=0.03）。LPS+抗 RAGE 组小鼠的 BALF/血清 FITC 比值低于 LPS+IgG 组，表明毛细血管通透性降低。定量 IHC RAGE 染色在 LPS+抗 RAGE ab 小鼠中低于 LPS+IgG 治疗小鼠（P=0.02）。