Page 16 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第41卷第2期
·166 · Journal of Nanjing Medical University（Natural Sciences） 2021年2月

·基础研究·

巨噬细胞中神经损伤诱导蛋白1在肝脏缺血再灌注损伤及其炎

症反应中的作用

周 顺，王 勇，张 峰，游 伟 *
南京医科大学第一附属医院肝胆中心，江苏 南京 210029

［摘 要］ 目的：评价巨噬细胞中神经损伤诱导蛋白1（nerve injury⁃induced protein 1，Ninj1）在肝脏缺血再灌注损伤及后续炎
症反应中的作用。方法：分别收集肝脏部分切除术中行肝门阻断的患者（肝门阻断组，n=6）以及未行肝门阻断的患者（对照
组，n=6）术前和术后的外周血，提取外周血单核细胞（peripheral blood mononuclear cell，PBMC），采用RT⁃PCR和Western blot方
法检测并比较两组患者术前、术后 PBMC 中 Ninj1 基因和蛋白表达水平；提取小鼠骨髓细胞，体外诱导成巨噬细胞（bone mar⁃
row derived macrophage，BMDM），检测脂多糖（lipopolysaccharide，LPS）刺激后 BMDM 中 Ninj1 表达的变化情况，并利用小干扰
RNA（small interfering RNA，siRNA）干扰 Ninj1 分析其对 BMDM 炎性反应的影响；建立小鼠肝脏缺血再灌注（ischemia reperfu⁃
sion，IR）模型，采用巨噬细胞特异性干扰 RNA 敲低巨噬细胞中 Ninj1 的表达，观察其对小鼠肝脏 IR 损伤及炎症反应的影响。
结果：Ninj1在肝门阻断组患者术后PBMC中的表达较术前明显增高（P < 0.05），而在对照组中无明显变化；体外LPS刺激能明
显激活小鼠BMDM中Ninj1的表达，siRNA干扰Ninj1的表达后LPS诱导的BMDM炎性反应明显减轻（P < 0.05）；在体内特异性
敲低巨噬细胞中 Ninj1 的表达能显著减轻小鼠肝脏 IR 损伤及炎症反应（P < 0.05）。结论：肝脏 IR 能激活人外周单核细胞中
Ninj1的表达。抑制巨噬细胞中Ninj1的表达能抑制小鼠肝脏IR后继发的炎性反应从而减轻肝脏的损伤。
［关键词］ 缺血再灌注；神经损伤诱导蛋白1；外周血单核细胞；巨噬细胞；炎症
［中图分类号］ R329.26 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）02⁃166⁃07
doi：10.7655/NYDXBNS20210203

Roles of macrophage nerve⁃injury induced protein1 in ischemia reperfusion⁃induced liver
injury and inflammatory response

ZHOU Shun，WANG Yong，ZHANG Feng，YOU Wei *
Hepatobiliary Center，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，China

［Abstract］ Objective：This study aims to evaluate the effect of Nerve injury ⁃ induced protein 1（Ninj1）in myeloid ⁃ derived
macrophages on liver ischemia/reperfusion（IR）⁃induced immune inflammatory response. Methods：Peripheral blood mononuclear cells
（PBMC）were collected from 6 patients who undergone hepatic portal occlusion（hepatic portal occlusion group）during hepatectomy
and another 6 patients who had no hepatic portal occlusion（control group）. RT⁃PCR and Western blot were performed to compare the
expression of Ninj1 in PBMC of pre⁃operation and post⁃operation between the two groups. Murine bone marrow derived macrophages
（BMDM）treated with lipopolysaccharide（LPS）were used to evaluate the role of Ninj1 in regulating macrophage inflammatory response
in vitro. A murine liver partial warm ischemia reperfusion model was established to determine the role of Ninj1 in liver IR injury and
hepatic inflammation. Results：In the hepatic portal occlusion group，the mRNA and protein levels of Ninj1 in PBMCs of post ⁃
operation were significantly higher than that in PBMC of pre⁃operation（P < 0.05），while no obvious change was observed in the control
group. Compared with the PBS group，Ninj1 was significantly upregulated in murine BMDM in response to LPS stimulation.
Knockdown of Ninj1 in BMDM by siRNA in vitro markedly reduced LPS⁃induced inflammatory cytokine secretion（P < 0.05）. Specific
blockade of Ninj1 in macrophages obviously attenuated liver IR injury and hepatic inflammation in vivo（P < 0.05）. Conclusion：Liver
IR significantly activated Ninj1 in patients’PBMCs. Inhibition of Ninj1 can mitigate macrophage inflammatory response leading to
attenuation of liver IR injury in mice.
［Key words］ ischemia reperfusion；nerve injury⁃induced protein 1；peripheral blood mononuclear cell；macrophage；inflammation
［基金项目］ 江苏省“六大人才高峰”高层次人才项目（WSW⁃019） ［J Nanjing Med Univ，2021，41（02）：166⁃172］
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通信作者（Corresponding author），E⁃mail：surgeonwu@163.com