脂肪组织来源的干细胞移植已被证明对治疗严重肝脏疾病有效。ADSC的预激活增强了其治疗效果。然而，这些影响尚未与胆汁淤积性肝损伤相关的研究进行过探究。本研究建立了一个胆汁淤积性肝损伤模型，采用胆管结扎（BDL）的方法在雄性C57BL/6小鼠中完成。通过尾静脉注射给小鼠人类ADSCs（hADSCs），预先进行肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）处理或未经处理。使用组织学染色、实时定量PCR（RT-qPCR）、Western blot和ELISA等方法评估hADSCs对BDL引起的肝损伤的疗效。在体外，研究了hADSCs条件培养基对肝星状细胞（HSCs）激活的影响。使用小干扰RNA（siRNA）靶向环氧合酶-2（COX-2）的研究方法，用于减轻hADSCs的活性。TNF-α/IL-1β预处理对免疫基因表达进行了调节，增强了hADSCs的移植效率。与对照hADSCs（C-hADSCs）相比，TNF-α/IL-1β预处理的hADSCs（P-hADSCs）显著缓解了BDL引起的肝损伤，表现为减轻肝细胞死亡、减轻Ly6G+中性粒细胞浸润和减少前炎性细胞因子TNF-α、IL-1β、C-X-C基序趋化因子配体1（CXCL1）和C-X-C基序趋化因子配体2（CXCL2）的表达。此外，P-hADSCs还明显延缓了BDL引起的肝纤维化的发展。在体外实验中，与C-hADSCs相比，P-hADSCs的条件培养基显著抑制了HSC活化。机械上，TNF-α/IL-1β上调了COX-2的表达，并增加了前列腺素E2（PGE2）的分泌。通过siRNA转染阻断COX-2，反转了P-hADSCs对PGE2产生、HSC活化和肝纤维化进展的益处。总之，我们的研究结果表明，TNF-α/IL-1β预处理增强了hADSCs在胆汁淤积性肝损伤小鼠中的疗效，部分通过COX-2 / PGE2途径实现。© 2023年。作者（S）保留所有权利。

Adipose tissue-derived stem cell (ADSC) transplantation has been shown to be effective for the management of severe liver disorders. Preactivation of ADSCs enhanced their therapeutic efficacy. However, these effects have not yet been examined in relation to cholestatic liver injury.In the present study, a cholestatic liver injury model was established by bile duct ligation (BDL) in male C57BL/6 mice. Human ADSCs (hADSCs) with or without tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) pretreatment were administrated into the mice via tail vein injections. The efficacy of hADSCs on BDL-induced liver injury was assessed by histological staining, real-time quantitative PCR (RT-qPCR), Western blot, and enzyme-linked immune sorbent assay (ELISA). In vitro, the effects of hADSC conditioned medium on the activation of hepatic stellate cells (HSCs) were investigated. Small interfering RNA (siRNA) was used to knock down cyclooxygenase-2 (COX-2) in hADSCs.TNF-α/IL-1β preconditioning could downregulate immunogenic gene expression and enhance the engraftment efficiency of hADSCs. Compared to control hADSCs (C-hADSCs), TNF-α/IL-1β-pretreated hADSCs (P-hADSCs) significantly alleviated BDL-induced liver injury, as demonstrated by reduced hepatic cell death, attenuated infiltration of Ly6G + neutrophils, and decreased expression of pro-inflammatory cytokines TNF-α, IL-1β, C-X-C motif chemokine ligand 1 (CXCL1), and C-X-C motif chemokine ligand 2 (CXCL2). Moreover, P-hADSCs significantly delayed the development of BDL-induced liver fibrosis. In vitro, conditioned medium from P-hADSCs significantly inhibited HSC activation compared to that from C-hADSCs. Mechanistically, TNF-α/IL-1β upregulated COX-2 expression and increased prostaglandin E2 (PGE2) secretion. The blockage of COX-2 by siRNA transfection reversed the benefits of P-hADSCs for PGE2 production, HSC activation, and liver fibrosis progression.In conclusion, our results suggest that TNF-α/IL-1β pretreatment enhances the efficacy of hADSCs in mice with cholestatic liver injury, partially through the COX-2/PGE2 pathway.© 2023. The Author(s).