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中华肝脏外科手术学电子杂志

中华肝脏外科手术学电子杂志 ›› 2025, Vol. 14 ›› Issue (01) : 118 -125. doi: 10.3877/cma.j.issn.2095-3232.2025010

基础研究

IGF-1 介导FOXO 信号通路在大鼠ALPPS 术后肝再生中的作用
蒙柄成1,2,3, 朱海2,3, 任洪冰1,2,3, 毛伟民1,2,3, 韦德令1,2,3, 徐邦浩1,2,3, 王继龙1,2,3, 金宗睿1,2,3, 蓝祝晶1,2,3, 黄柯豫1,2,3, 卢婷婷4, 张灵5, 郭雅2,3, 文张2,3,†()   
  1. 1.530021 南宁，广西医科大学
    2.530021 南宁，广西医科大学第一附属医院肝胆外科
    3.530021 南宁，广西消化道肿瘤加速康复外科基础研究重点实验室
    4.530021 南宁，广西医科大学第一附属医院超声科
    5.530021 南宁，广西医科大学第一附属医院放射科
  • 收稿日期:2024-10-22 出版日期:2025-02-10
  • 通信作者: 文张
  • 基金资助:
    国家自然科学基金地区基金项目（82160128）

Role of IGF-1-mediated FOXO signaling pathway in liver regeneration after ALPPS in rat models

Bingcheng Meng1,2,3, Hai Zhu2,3, Hongbing Ren1,2,3, Weimin Mao1,2,3, Deling Wei1,2,3, Banghao Xu1,2,3, Jilong Wang1,2,3, Zongrui Jin1,2,3, Zhujing Lan1,2,3, Keyu Huang1,2,3, Tingting Lu4, Ling Zhang5, Ya Guo2,3, Zhang Wen2,3,†()   

  1. 1.Guangxi Medical University, Nanning 530021, China
    2.Department of Hepatobiliary Surgery,the First Affiliated Hospital of Guangxi Medical University, Nanning 530021,China
    3.Key Laboratory of Basic Research of Enhanced Recovery after Surgery for Digestive Tract Tumors in Guangxi Zhuang Autonomous Region, Nanning 530021, China
    4.Department of Ultrasound, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021,China
    5.Department of Radiology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021,China
  • Received:2024-10-22 Published:2025-02-10
  • Corresponding author: Zhang Wen
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蒙柄成, 朱海, 任洪冰, 毛伟民, 韦德令, 徐邦浩, 王继龙, 金宗睿, 蓝祝晶, 黄柯豫, 卢婷婷, 张灵, 郭雅, 文张. IGF-1 介导FOXO 信号通路在大鼠ALPPS 术后肝再生中的作用[J/OL]. 中华肝脏外科手术学电子杂志, 2025, 14(01): 118-125.

Bingcheng Meng, Hai Zhu, Hongbing Ren, Weimin Mao, Deling Wei, Banghao Xu, Jilong Wang, Zongrui Jin, Zhujing Lan, Keyu Huang, Tingting Lu, Ling Zhang, Ya Guo, Zhang Wen. Role of IGF-1-mediated FOXO signaling pathway in liver regeneration after ALPPS in rat models[J/OL]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2025, 14(01): 118-125.

目的

探讨IGF-1 介导FOXO 信号通路在大鼠ALPPS 术后肝再生中作用。

方法

36 只雄性Wistar 大鼠SPF 环境中饲养，所有操作均符合实验动物伦理学要求。建立Wistar 大鼠ALPPS、门静脉结扎术（PVL）及假手术（Sham）模型。术后24、48、72 h 采用Ki-67 免疫组化染色，观察肝组织再生情况。术后72 h 的肝组织行转录组测序，并运用加权基因共表达网络分析（WGCNA）和KEGG富集分析等生物信息学方法筛选得到关键基因胰岛素样生长因子（IGF-1）。采用RT-qPCR 验证关键基因IGF-1 在大鼠肝脏的相对表达量。3 组间比较采用单因素方差分析和LSD-t 检验。

结果

免疫组化法检测显示，ALPPS 组 和PVL 组术后24、48、72 h 肝组织Ki-67 阳性细胞数逐渐增加，ALPPS 组在术后72 h 的 Ki-67 表达最高（F=927.28，P＜0.05），明显高于PVL 和Sham 组（LSD-t=13.45，42.15；P＜0.05）。WGCNA 分析将基因分为6 个模块，其中蓝色KMEblue 模块与ALPPS 相关性最高（r=0.69，P=0.002）。对蓝色模块（包含4 073 个基因）分析发现，IGF-1 基因在该模块内连接度最高。KEGG 富集分析揭示10 条相关通路，发现FOXO、P53 和细胞周期信号通路与肝再生密切相关，其中IGF-1 在FOXO 信号通路中的连接度最高。RT-qPCR 结果显示，与PVL 和Sham 组相比，ALPPS 组肝组织术后72 h 的IGF-1 mRNA 表达量明显升高（F=4.989，P＜0.05），明显高于PVL 和Sham 组（LSD-t=2.43，2.96；P＜0.05）。

结论

FOXO 信号通路中的IGF-1 可能介导大鼠ALPPS 术后肝再生过程。

关键词: 肝切除术, 联合肝脏离断和门静脉结扎的分阶段肝切除术（ALPPS）, 肝切除术后肝衰竭（PHLF）, 肝再生, 胰岛素样生长因子（IGF-1）, 信号通路

Objective

To investigate the role of FOXO signaling pathway mediated by IGF-1 in liver regeneration after the associating liver partition and portal vein ligation for staged hepatectomy(ALPPS) in rat models.

Methods

36 male Wistar rats were raised in SPF-grade laboratory environment,and all procedures met the requirements of laboratory animal ethics.Wistar rat models of ALPPS (ALPPS group), portal vein ligation (PVL group) and sham surgery (Sham group) were established.The regeneration of liver tissues at postoperative 24, 48 and 72 h was observed by Ki-67 immunohistochemical staining.At postoperative 72 h, transcriptome sequencing was performed in liver tissues.The key gene of insulinlike growth factor (IGF-1) was screened by bioinformatics methods such as weighted gene co-expression network analysis (WGCNA) and KEGG enrichment analysis.The relative expression of IGF-1 in rat liver was validated by RT-qPCR.Comparison among three groups was conducted by one-way ANOVA and LSD-t test.

Results

Immunohistochemical staining showed that the number of Ki-67 positive cells in liver tissues in the ALPPS and PVL groups was gradually increased at postoperative 24, 48 and 72 h, and the expression level of Ki-67 at postoperative 72 h was the highest in the ALPPS group (F=927.28, P＜0.05), significantly higher than those in the PVL and Sham groups (LSD-t=13.45, 42.15; both P＜0.05).The genes were divided into six modules by WGCNA, and the blue KMEblue module showed the highest correlation with ALPPS(r=0.69, P=0.002).Analysis of the blue module (containing 4 073 genes) showed that IGF-1 gene showed the highest connectivity in this module.KEGG enrichment analysis revealed 10 related signaling pathways, and found that FOXO, P53 and cell cycle signaling pathways were intimately correlated with liver regeneration.Among them, IGF-1 showed the highest connectivity in the FOXO signaling pathway.RT-qPCR indicated that compared with the PVL and Sham groups, the expression level of IGF-1 mRNA at postoperative 72 h was significantly up-regulated in the ALPPS group (F=4.989, P＜0.05), significantly higher than those in the PVL and Sham groups (LSD-t=2.43, 2.96; both P＜0.05).

Conclusions

IGF-1 in the FOXO signaling pathway may mediate liver regeneration after ALPPS in rats.

Key words: Hepatectomy, Associating liver partition and portal vein ligation for staged hepatectomy (ALPPS), Post-hepatectomy liver failure (PHLF), Liver regeneration, Insulin-like growth factor 1 (IGF-1), Signaling pathway
图1 大鼠ALPPS 模型 注：a 为结扎门静脉右外支、左外支和左中支的总支；b 为肝实质劈离后使用凝胶海绵止血分隔；RLL 为右外侧叶，RML 为右中叶，LML 为左中叶，LLL 为左外叶，CL 为尾状叶；实心箭头为右外叶门静脉支，虚线箭头为左外叶及左中叶门静脉总支
表1 IGF-1 引物与内参引物序列
基因 F R
Rat-IGF-1 TGAGCGCACCTCCAATAAAGAT TTGGTCCACACACGAACTGAAG
Rat-β-actin ATCCTGCGTCTGGACCTGG CCGCTCATTGCCGATAGTG
图2 三组大鼠术后肝脏Ki-67 免疫组化法检测结果 注：PVL 为门静脉结扎术，Sham 为假手术
图3 WGCNA 筛选与肝再生相关的模块与基因 注：a 为网络层次聚类，b 为最佳软阈值的选取，c 为模块基因聚类，d 为模块与肝再生的相关性热图，e 为模块与肝再生的聚类图，f 为TOM 拓扑网络图；TOM 为拓扑重叠矩阵，WGCNA 为加权基因共表达网络分析
图4 肝再生相关通路的关键模块内关键基因筛选与KEGG 富集分析 注：a 为肝再生最相关模块内基因显著性与高模块成员GS-MM 图，b 和c 为KEGG 富集分析，d 示 KEGG 富集分析基因
图5 肝再生相关通路的关键模块内关键基因筛选 注：a 为FOXO 信号通路内基因，b 为IGF-1 网络调控图；IGF-1 为胰岛素样生长因子
图6 qRT-PCR 检测三组大鼠肝脏中IGF-1 mRNA 表达水平 注：PVL 为门静脉结扎术，Sham 为假手术，IGF-1 为胰岛素样生长因子；*为P＜0.05
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