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厄贝沙坦减轻脂多糖诱导的肺损伤和.

Irbesartan eases lipopolysaccharide-induced lung injury and .

机构信息

Department of Pharmacy, Wuhan Red Cross Hospital, Wuhan, Hubei, China.

Department of Pharmacy, Yantai Yuhuangding Hospital, Yantai, Shandong, China.

出版信息

Chin J Physiol. 2023 Nov-Dec;66(6):516-525. doi: 10.4103/cjop.CJOP-D-23-00131.

Abstract

Acute lung injury (ALI) is classified as a devastating pulmonary disorder contributing to significant incidence and fatality rate. Irbesartan (IRB) is an angiotensin II receptor blocker that has been proposed to protect against oleic acid-induced ALI. To this end, the current study is concentrated on ascertaining the role of IRB in ALI and figuring out the probable action mechanism. First, cell counting kit-8 (CCK-8) appraised the viability of human pulmonary microvascular endothelial cells (HPMVECs) exposed to ascending concentrations of IRB. HPMVEC injury model and a mouse model of ALI induced by lipopolysaccharide (LPS) were pretreated by IRB. In vitro, cell viability was estimated by CCK-8 assay, and lactate dehydrogenase (LDH) release was tested by LDH assay kit. Enzyme-linked immunosorbent assay (ELISA) and Western blotting estimated the expression levels of inflammatory factors. Fluorescein isothiocyanate-dextran was used to assess HPMVEC permeability. Western blotting examined the expression of adherent and tight junction proteins. In vivo, hematoxylin and eosin staining evaluated lung tissue damage and lung wet/dry (W/D) weight was measured. ELISA analyzed the levels of inflammatory factors in the serum and bronchoalveolar lavage fluid (BALF), and Western blotting examined the expression of inflammatory factors. The total cell, neutrophil, and macrophage numbers in BALF were determined using a cell counter. Lung capillary permeability was assayed by Evans blue albumin and total protein concentration in BALF was measured using bicinchoninic acid method. Immunofluorescence assay and Western blotting examined the expression of adherent and tight junction proteins in lung tissues. It was observed that IRB dose-dependently enhanced the viability while reduced LDH release, inflammatory response as well as permeability in LPS-challenged HPMVECs in vitro. In addition, LPS-stimulated lung tissue damage, pulmonary edema, inflammatory response as well as lung capillary permeability in vivo were all reversed following IRB treatment. Collectively, IRB treatment might elicit protective behaviors against LPS-triggered ALI.

摘要

急性肺损伤 (ALI) 是一种破坏性的肺部疾病,其发病率和死亡率都很高。厄贝沙坦 (IRB) 是一种血管紧张素 II 受体阻滞剂,已被提议用于预防油酸诱导的 ALI。为此,本研究集中于确定 IRB 在 ALI 中的作用,并找出可能的作用机制。首先,通过细胞计数试剂盒-8 (CCK-8) 评估暴露于递增浓度的 IRB 的人肺微血管内皮细胞 (HPMVEC) 的活力。用 IRB 预处理 HPMVEC 损伤模型和脂多糖 (LPS) 诱导的小鼠 ALI 模型。在体外,通过 CCK-8 测定法评估细胞活力,通过 LDH 测定试剂盒测定乳酸脱氢酶 (LDH) 的释放。酶联免疫吸附测定 (ELISA) 和 Western blot 估计炎症因子的表达水平。荧光素异硫氰酸酯-葡聚糖用于评估 HPMVEC 通透性。Western blot 检查黏附连接和紧密连接蛋白的表达。在体内,苏木精和伊红染色评估肺组织损伤,并测量肺湿/干 (W/D) 重量。ELISA 分析血清和支气管肺泡灌洗液 (BALF) 中炎症因子的水平,Western blot 分析炎症因子的表达。使用细胞计数器测定 BALF 中的总细胞、中性粒细胞和巨噬细胞数量。通过 Evans 蓝白蛋白测定肺毛细血管通透性,并通过双缩脲法测量 BALF 中的总蛋白浓度。免疫荧光测定和 Western blot 检查肺组织中黏附连接和紧密连接蛋白的表达。结果表明,IRB 呈剂量依赖性地增强 LPS 刺激的 HPMVEC 活力,同时降低 LDH 释放、炎症反应和通透性。此外,IRB 治疗可逆转 LPS 刺激的肺组织损伤、肺水肿、炎症反应和肺毛细血管通透性。综上所述,IRB 治疗可能对 LPS 触发的 ALI 产生保护作用。

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