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  

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

作　　者：周顺 王勇[1] 张峰 游伟 ZHOU Shun;WANG Yong;ZHANG Feng;YOU Wei(Hepatobiliary Center,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院肝胆中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2021年第2期166-172,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省“六大人才高峰”高层次人才项目(WSW-019)。

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 marrow 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 reperfusion,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。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.

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 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]