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Updated: Mar 25, 2026
Cecal Ligation and Puncture-induced Sepsis as a Model To Study Autophagy in Mice
Published on: February 9, 2014
Effect of reticulocalbin 3 on monocyte-to-macrophage differentiation in sepsis through modulating autophagy
Danni Zan1,2, Zhuo Xu3, Fangping Ding3
1Medical Research Center, Beijing Institute of Respiratory Medicine and Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China.
Background:
Although monocyte-to-macrophage differentiation is essential for innate immune defense, its dysregulation can drive excessive inflammation in sepsis. Reticulocalbin 3 (RCN3) is an endoplasmic reticulum chaperone in the secretory pathway implicated in alveolar epithelial maturation and lung injury repair. This study aimed to investigate the effect of RCN3 on monocyte-to-macrophage differentiation in pneumonia-associated sepsis.
Methods:
Patients with pneumonic sepsis ( n = 10) and age- and sex-matched healthy volunteers ( n = 8) were enrolled from Beijing Chao-Yang Hospital. Peripheral blood was collected within 12 h after diagnosis; peripheral blood mononuclear cells (PBMCs) were isolated and monocytes were purified for transcriptional analyses of RCN3 and macrophage markers. For in vivo validation, myeloid-specific Rcn3 knockout mice received intratracheal lipopolysaccharide (10 mg/kg) or saline, and were assessed 16 h later ( n = 3-6 per group). Mechanistic studies in THP1 cells, human primary circulating monocytes and mouse bone marrow-derived monocytes used RCN3 knockdown and overexpression to evaluate differentiation markers, intracellular oxidative stress, and phagocytosis assays and autophagy flux evaluation.
Results:
In patients with pneumonic sepsis, RCN3 was significantly increased in circulating monocytes, but not lymphocytes, and its expression correlated with the macrophage marker CD68. RCN3 was also upregulated during monocyte-to-macrophage differentiation in both human and mouse monocytes. Furthermore, RCN3 knockdown attenuated differentiation, phagocytosis, and the lipopolysaccharide (LPS)-induced inflammatory response, whereas RCN3 overexpression had opposite effects. In vivo , myeloid-specific Rcn3 deletion decreased lung interstitial macrophage accumulation after intratracheal LPS instillation and alleviated acute lung injury. Mechanistically, the blunted differentiation caused by RCN3 deficiency was associated with reduced autophagy, due to impaired autophagosome formation.
Conclusions:
The upregulation of RCN3 in sepsis facilitates monocyte-to-macrophage differentiation by modulating autophagy. These findings also suggest that RCN3 in circulating monocytes presents a potential therapeutic target for sepsis.
