低温氧合灌注通过Brg1/Nrf2/HO-1轴减轻小鼠脂肪肝的缺血再灌注损伤。
Hypothermic oxygenated perfusion ameliorates ischemia-reperfusion injury of fatty liver in mice via Brg1/Nrf2/HO-1 axis.
作者信息
Wang Shengjie, Zeng Xianpeng, Yang Yunying, Li Shiyi, Wang Yanfeng, Ye Qifa, Fan Xiaoli
机构信息
Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Wuhan, China.
Department of Urology, Union Hospital, Affiliated TongJi Medical College, Huazhong University of Science & Technology, Wuhan, China.
出版信息
Artif Organs. 2022 Feb;46(2):229-238. doi: 10.1111/aor.14076. Epub 2021 Oct 13.
BACKGROUND
After cold storage (CS) and subsequent transplantation, fatty liver is more inclined to develop liver dysfunction and serious postoperative complications in contrast to healthy liver. Hypothermic oxygenated perfusion (HOPE) is a safe and efficacious system, which can repair fatty liver and reduce ischemia-reperfusion injury. The aim of this research is to investigate the function of Brg1/Nrf2/HO-1 signaling pathway in the protective effect of HOPE on ischemia-reperfusion injury of fatty liver.
METHODS
The mouse fatty liver model was successfully established and verified by hematoxylin-eosin (HE) staining and oil red O staining. The animals were divided into Control group, CS group and HOPE group. The levels of liver enzyme and lactate in the perfusate were used to measure liver function and cellular metabolism. HE staining and TUNEL staining were utilized to assess the tissue structure and apoptosis, respectively. The levels of superoxide dismutase, malondialdehyde and reactive oxygen species in liver tissue were measured to quantitatively analyze the degree of oxidative stress, and the expressions of protein Brg1, Nrf2 and HO-1 were detected by means of the western blot. Double-labeling immunofluorescence was to explore the colocalization of Brg1 and Nrf2.
RESULTS
The injury of the liver in the CS group was more serious than that in the control group. However, HOPE could significantly reduce the injury, which was manifested by the improvement of liver function and cellular metabolism, and the lower degrees of apoptosis, necrosis and oxidative stress. Furthermore, the expressions of Brg1, Nrf2 and HO-1 in the HOPE group were significantly increased than those in the CS group.
CONCLUSIONS
One-hour HOPE treatment before reperfusion can obviously improve the injury of fatty liver in mice. The underlying mechanism may be that the interaction of Brg1 and Nrf2 can selectively activate the transcription of HO-1.
背景
与健康肝脏相比，脂肪肝在冷藏（CS）及随后移植后更易发生肝功能障碍和严重的术后并发症。低温氧合灌注（HOPE）是一种安全有效的系统，可修复脂肪肝并减少缺血再灌注损伤。本研究旨在探讨Brg1/Nrf2/HO-1信号通路在HOPE对脂肪肝缺血再灌注损伤保护作用中的功能。
方法
通过苏木精-伊红（HE）染色和油红O染色成功建立并验证小鼠脂肪肝模型。将动物分为对照组、CS组和HOPE组。用灌注液中肝酶和乳酸水平来测量肝功能和细胞代谢。分别用HE染色和TUNEL染色评估组织结构和细胞凋亡。测量肝组织中超氧化物歧化酶、丙二醛和活性氧水平以定量分析氧化应激程度，并用蛋白质印迹法检测蛋白质Brg1、Nrf2和HO-1的表达。采用双标免疫荧光法探究Brg1和Nrf2的共定位。
结果
CS组肝脏损伤比对照组更严重。然而，HOPE可显著减轻损伤，表现为肝功能和细胞代谢改善，凋亡、坏死和氧化应激程度降低。此外，HOPE组中Brg1、Nrf2和HO-1的表达明显高于CS组。
结论
再灌注前1小时的HOPE治疗可明显改善小鼠脂肪肝损伤。潜在机制可能是Brg1和Nrf2的相互作用可选择性激活HO-1的转录。
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