羟基红花黄色素A通过抑制坏死性凋亡减轻小鼠热射病诱导的急性肺损伤
[Hydroxysafflor yellow A attenuates heat stroke-induced acute lung injury in mice by inhibiting necroptosis].
作者信息
Liu X, Zhang S, Yu R, Lin X, Fan W, Wang Y, Liang Z, Xie W, Liu Y, Chen H
机构信息
Department of Critical Care Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Department of Critical Care Medicine, Jiangmen Central Hospital, Jiangmen 529000, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Dec 20;42(12):1875-1881. doi: 10.12122/j.issn.1673-4254.2022.12.17.
OBJECTIVE
To investigate the protective effect of hydroxysafflor yellow A (HSYA) against heat stroke (HS)-induced acute lung injury and its possible mechanism.
METHODS
The optimal dose of HSYA pretreatment intraperitoneal injection prior to HS was determined in a mice by observing heat tolerance of the mice. C57BL/6J mice were pretreated with HSYA at the optimal dose or with Nec-1 (a RIP1 activation inhibitor) before HS, and the changes in core body temperature and survival of the mice were observed during the 72-h recovery period. At different stages of recovery, lung tissues, bronchoalveolar lavage fluid and blood samples were collected from the mice for assessing lung tissue pathology, wet-to-dry weight ratio and water content of the lungs; leukocyte and neutrophil counts, total protein levels and HMGB1 level in the bronchoalveolar lavage fluid (BLF) were also detected. Serum levels of TNF-α, IL-6 and HMGB1 were detected with ELISA, and the expression levels of RIP1, RIP3, MLKL-s358, MLKL and MLKL-s358 proteins in the lung tissues were detected using Western blotting.
RESULTS
HSYA pretreatment at the moderate and high doses significantly improved heat tolerance of the mice with comparable effects. At the optimal dose of 2.25 mg/kg, HSYA pretreatment significantly increased heat tolerance of the mice (<0.05), showing a similar effect with Nec-1 pretreatment. Pretreatment with HSYA and Nec-1 both significantly increased survival rate of the mice (<0.05), lowered histopathological score and water content of the lungs, and reduced the levels of TNF-α, IL-6 and HMGB1 (<0.05), leukocyte and neutrophil counts, and total protein and HMGB1 levels in the BLF (<0.05). The mice during recovery from HS showed significantly increased RIP1 expression and MLKL-s358 phosphorylation level in the lung tissue (<0.05), which were obviously lowered by HSYA pretreatment of the mice.
CONCLUSION
Severe HS results in necroptosis in the lung tissue of mice, which can be alleviated by HSYA pretreatment.
目的
探讨羟基红花黄色素A（HSYA）对热射病（HS）诱导的急性肺损伤的保护作用及其可能机制。
方法
通过观察小鼠的耐热性，确定HS前腹腔注射HSYA预处理的最佳剂量。C57BL/6J小鼠在HS前用最佳剂量的HSYA或Nec-1（一种RIP1激活抑制剂）进行预处理，并在72小时恢复期内观察小鼠的核心体温变化和存活率。在恢复的不同阶段，从小鼠采集肺组织、支气管肺泡灌洗液和血液样本，以评估肺组织病理学、肺湿干重比和含水量；还检测支气管肺泡灌洗液（BLF）中的白细胞和中性粒细胞计数、总蛋白水平和HMGB1水平。用ELISA检测血清TNF-α、IL-6和HMGB1水平，用蛋白质印迹法检测肺组织中RIP1、RIP3、MLKL-s358、MLKL和MLKL-s358蛋白的表达水平。
结果
中、高剂量的HSYA预处理均能显著提高小鼠的耐热性，效果相当。在最佳剂量2.25mg/kg时，HSYA预处理显著提高了小鼠的耐热性（<0.05），与Nec-1预处理效果相似。HSYA和Nec-1预处理均显著提高了小鼠的存活率（<0.05），降低了肺组织病理学评分和含水量，降低了TNF-α、IL-6和HMGB1水平（<0.05）、白细胞和中性粒细胞计数以及BLF中的总蛋白和HMGB1水平（<0.05）。HS恢复过程中的小鼠肺组织中RIP1表达和MLKL-s358磷酸化水平显著升高（<0.05），而HSYA预处理可明显降低。
结论
严重HS可导致小鼠肺组织坏死性凋亡，HSYA预处理可减轻这种损伤。
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