诊断学理论与实践

诊断学理论与实践 ›› 2025, Vol. 24 ›› Issue (01): 35-42.doi: 10.16150/j.1671-2870.2025.01.006

• 论著 • 上一篇    下一篇

外周血Scimp可作为代谢相关脂肪性肝病肝硬化的生物诊断标志物——基于动物实验的研究

陈洪卫1,2, 朱婷1, 刘燕1, 侯彦强2, 范广建1()   

  1. 1.上海交通大学医学院附属第一人民医院临床研究院难治性疾病精准研究中心，上海 200080
    2.上海交通大学医学院附属松江医院检验科，上海 201600
  • 收稿日期:2024-10-15 接受日期:2024-12-31 出版日期:2025-02-25 发布日期:2025-02-05
  • 通讯作者: 范广建 E-mail：gjfan@shsmu.edu.cn
  • 基金资助:
    上海市松江区科学技术攻关项目(22SJKJGG73);上海市松江区公共卫生体系建设三年行动计划项目(SJGW6-28);上海市医学重点学科(2024ZDXK0065)

Study on exploring Scimp in peripheral blood as a biodiagnostic marker for MAFLD-related cirrhosis based on animal experiments

CHEN Hongwei1,2, ZHU Ting1, LIU Yan1, HOU Yanqiang2, FAN Guangjian1()   

  1. 1. Precision Research Center for Refractory Diseases, Institute for Clinical Research, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine. ;Shanghai 200080, China
    2. Department of Laboratory Medicine, Songjiang Hospital Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai 201600, China
  • Received:2024-10-15 Accepted:2024-12-31 Published:2025-02-25 Online:2025-02-05

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摘要：

目的 寻找代谢相关脂肪性肝病（metabolic associated fatty liver disease，MAFLD）进展为肝硬化的诊断生物标志物，为肝硬化疾病的早诊、早治提供诊断依据。方法 选用健康雄性SD（Sprague Dawley）大鼠，随机分为正常对照（normal control，NC）组（15只）、高脂饮食（high-fat diet，HFD）组（15只）和肝纤维化（liver fibrosis，LF）组（15只）。NC组大鼠接受普通饲料喂养，HFD组持续高脂饲料喂养以诱导脂肪肝和轻度肝纤维化，LF组持续高脂饲料喂养同时注射四氯化碳诱导肝硬化。饲养41周后，采集大鼠血液和肝脏组织。肝脏组织采用苏木精-伊红（hematoxylin and eosin，HE）染色、油红O染色和天狼星红染色，以评价肝脏组织形态、脂肪浸润及肝纤维化程度。利用mRNA测序技术分析血液样本筛选差异表达基因，采用反转录实时荧光定量聚合酶链式反应（reverse transcription quantitative real-time polymerase chain reaction， RT-qPCR）技术验证测序筛选的差异基因表达情况。另收集2025年1月至2月来自上海交通大学医学院附属松江医院的健康体检者、MAFLD和肝硬化患者的血液样本各10例，验证筛选出的差异基因的mRNA表达情况。结果 大鼠外周血细胞mRNA测序筛选出17个差异基因，包括Rab3ip、Gprasp2、Emid1、Hbq1b、Scimp、Wipf3、Scrn1、Sez6、Bglap、Bhlhb9、Ranbp10、Ubd、Plekhb1、Nup210l、Gp1bb、Cpne8和Oscar；RT-qPCR验证结果表明，Scimp基因在NC组、HFD组、LF组的mRNA表达水平依次呈上调趋势，与测序结果一致，各组间两两比较，差异有统计学意义（P<0.01）。人外周血细胞RT-qPCR验证结果与大鼠外周血Scimp基因的mRNA测序和RT-qPCR验证表达趋势一致。在体检人群、MAFLD患者及肝硬化患者间，Scimp存在同样的表达差异（P<0.05）。结论 外周血细胞Scimp基因的mRNA表达水平可作为早期肝硬化的潜在无创诊断标志物。

关键词: 代谢相关脂肪性肝病, 肝硬化, 诊断标志物, Scimp基因

Abstract:

Objective This study aims to identify biodiagnostic markers for the progression of metabolic-associated fatty liver disease (MAFLD) to cirrhosis, providing evidence for early diagnosis and treatment of cirrhosis. Methods The experiment utilized healthy male Sprague Dawley (SD) rats, which were randomly divided into a normal control (NC) group of 15 rats, a high-fat diet (HFD) group of 15 rats, and a liver fibrosis (LF) group of 15 rats. Rats in the NC group were fed a normal diet, those in the HFD group received continuous high-fat diet to induce fatty liver and mild liver fibrosis, and those in the LF group were fed a high-fat diet and injected with carbon tetrachloride to induce cirrhosis. After 41 weeks of feeding, blood and liver tissues were collected from rats. Liver tissues were stained with hematoxylin and eosin (HE), Oil Red O, and Sirius red to evaluate liver morphology, fat infiltration, and liver fibrosis. Differentially expressed genes in blood samples were screened using mRNA sequencing technology, and the expression of sequencing-screened differentially expressed genes was validated by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Additionally, blood samples from 10 human subjects each in the NC, MAFLD, and cirrhosis groups were collected from January to February 2025 at Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine to validate the mRNA expression of screened differentially expressed genes. Results The mRNA sequencing of peripheral blood cells in rats identified 17 differentially expressed genes, including Rab3ip, Gprasp2, Emid1, Hbq1b, Scimp,Wipf3, Scrn1, Sez6, Bglap, Bhlhb9, Ranbp10, Ubd, Plekhb1, Nup210l, Gp1bb, Cpne8, and Oscar. The RT-qPCR validation results showed that the mRNA expression levels of the Scimp gene in the NC group, HFD group, and LF group exhibited an increasing trend in sequence, which was consistent with the sequencing results. Pairwise comparisons between groups showed statistically significant differences (P < 0.01). The RT-qPCR validation results of human peripheral blood cells were consistent with the mRNA sequencing and RT-qPCR validation expression trends of the Scimp gene in rat peripheral blood. Scimp showed the same expression differences among NC, MAFLD, and cirrhosis patients (P < 0.05). Conclusions The mRNA expression level of the Scimp gene in peripheral blood cells can serve as a potential non-invasive biodiagnostic marker for early-stage cirrhosis.

Key words: Metabolic-associated fatty liver disease, Cirrhosis, Biodiagnostic marker, Scimp gene

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