敲低 HO-1 基因对小鼠脾脏和脓毒症急性肺损伤组织中免疫细胞的影响

Effects of Heme Oxygenase-1 (HO-1) Gene Knockdown on Immune Cells in the Spleen and Septic Acute Lung Injury Tissues of Mice

摘要

为了探究血红素加氧酶-1（HO-1）基因对小鼠脾脏和脓毒症急性肺损伤组织中免疫细胞的影响，本研究构建了HO-1 ^{+/ -}杂合子基因型小鼠模型。通过该模型，我们研究了HO-1基因对脾脏和急性肺损伤组织中免疫细胞的调控作用。在小鼠脓毒症模型中，采用盲肠结扎穿孔法（CLP）诱导脓毒症，术后24 h通过流式细胞术和HE染色检测脾脏及肺损伤组织中免疫细胞的变化。结果显示：与对照组相比，HO-1基因敲低导致脾脏组织中CD3-NK1.1 ⁺和CD3 ⁺ CD4 ⁺细胞比例显著降低（ P<0.05、 P<0.01），而CD3 ⁺ CD8 ⁺和调节性T细胞（Treg）比例显著升高（ P<0.05、 P<0.01）。此外，HO-1基因敲低加剧了CLP小鼠的急性肺损伤，肺组织中红细胞数量、肺泡壁厚度、中性粒细胞、肺损伤评分均升高，且具有统计学意义（ P<0.05），且肺组织中巨噬细胞比例增加（ P<0.05）并向M1型巨噬细胞极化。因此，本研究发现HO-1敲低导致脾脏T细胞亚群失衡及肺部巨噬细胞向M1型极化偏移，进而加剧脓毒症诱导的急性肺损伤。这揭示了HO-1通过协调脾-肺免疫细胞功能以减轻器官损伤的关键保护机制。

Abstract

To investigate the effect of heme oxygenase-1 (HO-1) gene on immune cells in mouse spleen and sepsis-induced acute lung injury tissues, this study constructs a HO-1 ^{+/ -} heterozygous mouse model. Using this model, it examines the regulatory role of HO-1 gene on immune cells in spleen and acute lung injury tissues. In the mouse sepsis model, sepsis is induced by cecal ligation and puncture (CLP). Twenty‑four hours after the procedure, flow cytometry and HE staining are performed to detect changes in immune cells in spleen and lung injury tissues. The results show that compared with the control group, HO-1 gene knockdown significantly decreases the proportions of CD3-NK1.1 ⁺ and CD3 ⁺ CD4 ⁺ cells in spleen tissue ( P<0.05, P<0.01), while significantly increases the proportions of CD3 ⁺CD8 ⁺ cells and regulatory T cells (Treg) ( P<0.05, P<0.01). In addition, HO-1 gene knockdown aggravates acute lung injury in CLP mice, with statistically significant increases in red blood cell count, alveolar wall thickness, neutrophil infiltration, and lung injury score in lung tissue ( P<0.05). Moreover, the proportion of macrophages in lung tissue increases (P<0.05) and polarizes toward M1-type macrophages. Therefore, this study discovers that HO-1 knockdown leads to an imbalance in splenic T-cell subsets and a polarization shift of pulmonary macrophages toward the M1 phenotype, thereby exacerbating sepsis-induced acute lung injury. This reveals the crucial protective mechanism by which HO-1 alleviates organ damage through coordinating spleen-lung immune cell functions.

关键词

血红素加氧酶-1 / 脓毒症 / 急性肺损伤 / 脾脏 / 免疫细胞

Key words

Heme oxygenase-1 / sepsis / acute lung injury / spleen / immune cells

参考文献

[1] Li X Y, Qiu C M, Yang F Y, et al. Protective effects of Prussian blue nanozyme against sepsis-induced acute lung injury by activating HO-1[J]. European Journal of Pharmacology, 2024, 968: 176354.

[2] Zhuang C, Kang M, Lee M . Delivery systems of therapeutic nucleic acids for the treatment of acute lung injury/acute respiratory distress syndrome[J]. Journal of Controlled Release, 2023, 360: 1-14.

[3] Xia Q, Liu G, Zhang L, et al. Anemonin suppresses sepsis-induced acute lung injury by inactivation of nuclear factor-kappa B and activation of nuclear factor erythroid 2-related factor-2/heme oxygenase-1 pathway[J]. Federation of American Societies for Experimental Biology Journal, 2025, 39(2): e70328.

[4] Jiang L, Jiang Q, Yang S, et al. GYY4137 attenuates LPS-induced acute lung injury via heme oxygenase-1 modulation[J]. Pulm Pharmacol Ther, 2019, 54: 77-86.

[5] Chen W, Zhang Y, Chen J, et al. Heme oxygenase-1 modulates macrophage polarization through endothelial exosomal miR-184-3p and reduces sepsis-induce lung injury[J]. International Journal of Nanomedicine, 2025, 20: 5039-5057.

[6] Li X, Yu J, Gong L, et al. Heme oxygenase-1(HO-1) regulates Golgi stress and attenuates endotoxin-induced acute lung injury through hypoxia inducible factor-1α (HIF-1α)/HO-1 signaling pathway[J]. Free Radical Biology and Medicine, 2021, 165: 243-253.

[7] Lu Z, Zhan F, Yang M, et al. The immune function of heme oxygenase-1 from grass carp (Ctenopharyngodon idellus) in response to bacterial infection [J]. Fish Shellfish Immunol, 2021, 112: 168-178.

[8] Yao L, Hao Y, Wen G, et al. Induction of heme oxygenase-1 modifies the systemic immunity and reduces atherosclerotic lesion development in ApoE deficient mice[J]. Front Pharmacol, 2022, 13: 809469.

[9] Regev D, Surolia R, Karki S, et al. Heme oxygenase-1 promotes granuloma development and protects against dissemination of mycobacteria[J]. Laboratory Investigation, 2012, 92(11): 1541-1552.

[10] Campbell N K, Fitzgerald H K, Dunne A . Regulation of inflammation by the antioxidant haem oxygenase 1[J]. Nature Reviews Immunology, 2021, 21(7): 411-425.

[11] Ozen M, Zhao H, Kalish F, et al. Heme oxygenase-1 deficiency results in splenic T-cell dysregulation in offspring of mothers exposed to late gestational inflammation[J]. American Journal of Reproductive Immunology, 2018, 79(5): e12829.

[12] Cuenca A G, Delano M J, Kelly-Scumpia K M, et al. A paradoxical role for myeloid-derived suppressor cells in sepsis and trauma[J]. Molecular Medicine, 2011, 17(3-4): 281-292.

[13] Liu C, Song W, Wu S, et al. Implications of protein posttranslational modifications in sepsis-induced Acute lung injury and acute respiratory distress syndrome[J]. Shock, 2025, 64(5): 454-465.

[14] Matthay M A, Zemans R L, Zimmerman G A, et al. Acute respiratory distress syndrome[J]. Nature Reviews Disease Primers, 2019, 5(1): 18.

[15] Shutong L, Yu J, Jia W, et al. HO-1/autophagic flux axis alleviated sepsis-induced acute lung injury via inhibiting NLRP3 inflammasome[J]. Cellular Signalling, 2022, 100: 110473.

[16] Bai X, Liu Y, Liu J, et al. ADSCs-derived exosomes suppress macrophage ferroptosis via the SIRT1/NRF2 signaling axis to alleviate acute lung injury in sepsis[J]. International Immunopharmacology, 2025, 146: 113914.

[17] Wegiel B, Hedblom A, Li M, et al. Heme oxygenase-1 derived carbon monoxide permits maturation of myeloid cells[J]. Cell Death and Disease, 2014, 5(3): e1139.

[18] Zhang M, Nakamura K, Kageyama S, et al. Myeloid HO-1 modulates macrophage polarization and protects against ischemia-reperfusion injury[J]. JCI Insight, 2018, 3(19): e120596.

[19] Shi B, Hao Y, Li W, et al. TIPE2 may target the Nrf2/HO-1 pathway to inhibit M1 macrophage-related neutrophilic inflammation in asthma[J]. Frontiers in Immunology, 2022, 13: 883885.

基金资助

山东省自然科学基金创新发展联合基金项目“多组学驱动的基于氢/氧混合气体治疗脓毒症肺损伤的靶点发现及新药筛选的研究”(ZR2024LSW011)

国家自然科学基金青年科学基金项目“内质网应激信号通路在氧化低密度脂蛋白调控内皮祖细胞功能和凋亡中的作用及机制”(816003603)

山东省自然科学基金重点项目“氢/氧混合吸入调控内皮祖细胞归巢治疗急性呼吸窘迫综合征的分子机制”(ZR2020KH008)