GPNMB 作为代谢相关脂肪性肝病及肝炎生物标志物的诊断与预测价值

GPNMB as a diagnostic and predictive biomarker for metabolic dysfunction-associated steatotic liver disease and steatohepatitis

摘要

目的 探究在代谢功能障碍相关脂肪性肝病 (MASLD) 和代谢功能障碍相关脂肪性肝炎 (MASH) 条件下, 非转移性黑色素瘤糖蛋白 B (GPNMB) 作为疾病发生进展的生物标志物的有效性。方法 通过分析 MASH 模型小鼠转录组数据及单细胞转录组数据, 评估在疾病模型下全肝及巨噬细胞的 GPNMB 表达情况。 将 20 只 8 周龄 C57BL/6 雄性小鼠随机分为 4 组, 分别为胆碱缺乏的氨基酸 (CDAA) 对照组、CDAA 饮食组、 高脂饮食 (HFD) 对照组和 HFD 组。 造模结束后取肝组织检测, 在 mRNA 及蛋白水平进行验证; ELISA 测定 MASH 模型小鼠血清中 GPNMB 的分泌表达; 体外诱导巨噬细胞分别向 M1 及 M2 型极化后 mRNA 水平检测 GPNMB 表达变化。结果 动物实验检测结果和测序数据分析结果高度一致, GPNMB 在对照组低表达, 但在疾病模型下的肝组织中 mRNA 和蛋白水平表达均上调 (P<0.01), 且主要在巨噬细胞表达; 造模后血清中 GPNMB 分泌增加 (P<0.05); GPNMB 的表达受巨噬细胞极化状态的调控。结论 GPNMB 有望成为诊断治疗 MASLD 和 MASH 的生物标志物。

Abstract

Objective To evaluate the effectiveness of glycoprotein non-metastatic melanoma protein B (GPNMB) as a biomarker for the occurrence and progression of metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis (MASH). Methods GPNMB expression changes in whole liver and macrophages under the disease model were assessed by analyzing RNA-sequencing and single-cell RNA-sequencing data from a mouse MASH model. Meanwhile, twenty 8-week-old male C57BL/6 mice were randomly assigned to four groups: choline-deficient L-amino acid-defined (CDAA), CDAA diet group, high-fat diet (HFD) control group, and HFD group. Liver samples harvested at the experimental endpoint were assessed for GPNMB expression at mRNA and protein levels. The secretion and expression of GPNMB in the serum of MASH model mice were determined by ELISA. Following in vitro induction of macrophage polarization towards M1 and M2 phenotypes, changes in GPNMB expression were examined at the mRNA level. Results Results from animal experiments closely aligned with sequencing data analyses. GPNMB was lowly expressed in the control group, but demonstrated significant upregulation at both mRNA and protein levels in liver tissues of disease models (P<0.01), predominantly localized to macrophages. After modeling, the secretion of GPNMB in the serum increased (P<0.05). The expression of GPNMB was modulated by the polarization state of macrophages. Conclusion GPNMB is expected to become a biomarker for the diagnosis and treatment of MASLD and MASH.

关键词

非转移性黑色素瘤糖蛋白 B / 代谢功能障碍相关脂肪性肝病 / 代谢功能障碍相关脂肪性肝炎 / 脂代谢 / 转录组测序 / 单细胞转录组测序 / 巨噬细胞极化 / 生物标志物

Key words

glycoprotein non-metastatic melanoma protein B / metabolic dysfunction-associated steatotic liver disease / metabolic dysfunction-associated steatohepatitis / lipid metabolism / RNA-seq / scRNA-seq / macrophage polarization / biomarker

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基金资助

国家自然科学基金杰出青年科学基金(82325007)