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目的 探究SAC1样磷脂酰肌醇磷酸磷酸酶（SAC1）调控巨噬细胞核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体激活的分子机制及其在代谢功能障碍相关脂肪性肝炎（MASH）病理进程中的作用。方法 利用单细胞RNA测序（scRNA-seq）分析对照组和MASH小鼠肝脏巨噬细胞亚群中SACM1L的表达水平差异。分离并培养对照组和高脂饮食组小鼠骨髓来源巨噬细胞（BMDMs），通过RT-qPCR及Western blot检测SACM1L和NLRP3的表达水平。运用CRISPR/Cas9技术分别构建SACM1L敲除的THP-1巨噬细胞系和HeLa细胞系，比较野生型与SACM1L敲除THP-1细胞中p65磷酸化及NLRP3炎症小体的活化状态；运用免疫荧光实验评估野生型与SACM1L敲除的HeLa细胞中分散的反式高尔基体网络（dTGN）对NLRP3的空间募集能力。此外，通过给予小鼠10周含0.1%蛋氨酸的胆碱缺乏高脂饮食（CDA-HFD）建立MASH模型，比较对照组与MASH小鼠肝脏组织中SACM1L与NLRP3的表达水平。结果 生信分析与体内实验一致表明，SACM1L与NLRP3在MASH小鼠的肝脏组织及巨噬细胞中表达均显著上调。敲除SACM1L可减弱三棕榈酰化半胱氨酰丝氨酸四赖氨酸（Pam3CSK4）诱导的p-p65水平的升高，抑制NLRP3向dTGN的空间募集，降低下游Caspase-1和IL-1β的成熟与释放。结论 SAC1通过促进p65信号活化促进NLRP3的表达，影响NLRP3向dTGN募集，促进巨噬细胞炎症反应，进而加速MASH的病理进程。
Abstract：Objective To investigate the molecular mechanism by which SAC1-like phosphatidylinositide phosphatase(SAC1) regulates NOD-like receptor protein 3(NLRP3) inflammasome activation in macrophages and to explore its role in the pathological progression of metabolic dysfunction-associated steatohepatitis(MASH).Methods Single-cell RNA sequencing(scRNA-seq) was utilized to analyze the differential expression levels of SACM1L in liver macrophage subpopulations between control and MASH mice. Bone marrow-derived macrophages(BMDMs) from control and high-fat diet-fed mice were isolated and cultured, and the expression levels of SACM1L and NLRP3 were determined via RT-qPCR and Western blot. CRISPR/Cas9 technology was employed to establish SACM1L-knockout THP-1 macrophage and HeLa cell lines. The phosphorylation of p65 and the activation status of the NLRP3 inflammasome were compared between wild-type and SACM1L-knockout THP-1 cells. Immunofluorescence assays were conducted to evaluate the spatial recruitment capacity of NLRP3 to the dispersed trans-Golgi network(dTGN) in wild-type and SACM1L-knockout HeLa cells. Furthermore, a MASH mouse model was established by feeding mice a choline-deficient high-fat diet containing 0. 1% methionine(CDA-HFD) for 10 weeks, and the expression levels of SACM1L and NLRP3 in liver tissues were compared between the control and MASH groups.Results Bioinformatics analysis and in vivo experiments consistently demonstrated that the expression levels of SACM1L and NLRP3 were significantly upregulated in both liver tissues and macrophages of MASH mice. SACM1L deficiency attenuated the increase in p-p65 levels induced by tripalmitoyl cysteinyl seryl tetra-lysine(Pam3 CSK4), inhibited the spatial recruitment of NLRP3 to the dTGN, and attenuated the subsequent maturation and release of downstream Caspase-1 and IL-1β.Conclusion SAC1 promotes NLRP3 expression by facilitating p65 signaling activation and mediates the spatial recruitment of NLRP3 to the dTGN. This process exacerbates the macrophage inflammatory response, thereby accelerating the pathological progression of MASH.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.08.001
中图分类号:R575
引用信息:
[1]孙志阳,曹新旺.SAC1介导的巨噬细胞NLRP3炎症小体激活在MASH炎症进展中的作用[J].安徽医科大学学报,2026,61(08):1333-1341.DOI:10.19405/j.cnki.issn1000-1492.2026.08.001.
基金信息:
国家自然科学基金项目（编号：91854120）~~
2026-07-08
2026-07-08
2026-07-08