In Vitro Cellular & Developmental Biology. Animal最新文献

{"title":"The abnormal thyroxine signals triggers podocyte apoptosis in DN mice.","authors":"Jing Wang, Binfang Huang, Guangquan Zhou","doi":"10.1007/s11626-026-01153-3","DOIUrl":"10.1007/s11626-026-01153-3","url":null,"abstract":"<p><p>Podocyte injury is a pivotal factor in the advancement of diabetic nephropathy (DN). The present study aimed to delineate the influence of disrupted thyroxine signaling on podocyte apoptosis in DN mouse models. We employed bioinformatics analyses, coupled with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment assessments, to identify differentially expressed genes (DEGs) associated with thyroxine signaling in both human and murine DN datasets. Subsequently, we elucidated the function of thyroid hormone receptor α1 (THRA1) and nuclear receptor co-repressor 1 (NCOR1) on glomerular injury and podocytes apoptosis under hypothyroid and hyperglycemic conditions, respectively. Our findings highlight that hypothyroidism significantly alters glomerular gene expression profiles in DN mice leading to increased podocyte apoptosis. This effect occurs through a dual mechanism: on one hand, the upregulation of THRA1 expression induced by DN results in direct glomerular injury, which was further aggravated by hypothyroidism; on the other hand, the downregulation of NCOR1 expression thereby increases THRA1 activity levels. Our data suggests that disturbed thyroxin signals could trigger podocyte apoptosis and glomerular injury in DN mice, offering new insights into DN pathogenesis while laying groundwork for innovative therapeutic strategies.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"444-458"},"PeriodicalIF":1.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of ARL4C antisense oligonucleotide with reduced off-target effects and enhanced efficacy as an anti-cancer drug.","authors":"Kanae Kawai, Shinji Matsumoto, Akikazu Harada, Ryota Sada, Takeshi Harada, Yuuya Kasahara, Satoshi Obika, Yasushi Okamura, Akira Kikuchi","doi":"10.1007/s11626-026-01154-2","DOIUrl":"10.1007/s11626-026-01154-2","url":null,"abstract":"<p><p>ARL4C, a member of the small GTP-binding protein superfamily, is overexpressed in the tumor lesions of various cancers. Antisense oligonucleotide (ASO) therapy targeting ARL4C (ASO-1316-A), a 15-mer ASO, has been shown to inhibit xenograft tumor formation. To enhance therapeutic efficacy and reduce off-target effects, fourteen 18-mer ARL4C-targeting ASOs were developed. Among them, ASO-2025-A/L demonstrated superior suppression of ARL4C mRNA and protein expression compared to ASO-1316-A. In vitro, ASO-2025-A/L inhibited the proliferation, migration, and adhesion abilities of PANC-1 and S2-CP8 pancreatic cancer cells as well as PC-9 lung cancer cells more effectively than ASO-1316-A. ASO-2025-A/L was predicted to reduce the expression of 26 potential off-target genes, compared to 2824 potential off-target genes affected by ASO-1316-A in PANC-1 cells. While ASO-2025-A/L downregulated 317 genes, which were confirmed by RNA-sequence analysis, the majority was involved in cell adhesion pathways downstream of ARL4C signaling, and only two genes corresponded to off-target genes. In vivo, intravenous administration of ASO-2025-A/L inhibited xenograft tumor growth induced by PC-9 and PANC-1 cells, accompanied by a reduction in ARL4C mRNA levels in tumors without elevating serum liver toxicity markers. These findings highlight the development of an improved 18-mer ARL4C-targeting ASO with enhanced therapeutic efficacy and reduced off-target effects.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"413-427"},"PeriodicalIF":1.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147369162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"C-Jun NH2-terminal kinase inhibitor bentamapimod prevents RANKL-mediated osteoclastogenesis and ovariectomy-induced osteoporosis.","authors":"Junming Huang, Mingchao Lin, Kui Deng, Song Zhou","doi":"10.1007/s11626-025-01147-7","DOIUrl":"10.1007/s11626-025-01147-7","url":null,"abstract":"<p><p>Over the past five decades, numerous treatment approaches have been developed to target either the inhibition of bone resorption or the promotion of bone formation. However, therapeutic strategies for osteoporosis still face limitations and safety concerns. Bentamapimod, initially designed for neurological disorders, is a novel JNK inhibitor that has demonstrated anti-tumor and immunomodulatory properties. In this study, we aim to investigate the protective role of bentamapimod against bone loss in mice suffering from estrogen deficiency. In vivo, we investigated that bentamapimod alleviated bone loss in mice suffering from estrogen deficiency. According to the micro-CT and histomorphometry assays, bentamapimod inhibits bone resorption as well as bone formation, but the effect of bentamapimod on bone resorption is better than that of bone formation, which ultimately results in ameliorating bone loss caused by ovariectomy. Moreover, we confirmed that bentamapimod can attenuate receptor activator of nuclear factor-kB ligand (RANKL)-induced osteoclast differentiation via inactivating the JNK, while anisomycin (a JNK agonist) partially confronts this effect. Our study demonstrated that bentamapimod would be able to be used as a promising new drug strategy for osteoporosis via inhibiting osteoclast differentiation both in vitro and in vivo.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"476-488"},"PeriodicalIF":1.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146213255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Network pharmacology analysis and experimental validation of modified Taohong Siwu decoction against atherosclerosis.","authors":"Changle Shao, Wanting Meng, Ke Ning, Mengying Huang, Han Li, Hao Cai, Yanwu Xu, Tao Zuo, Hai-Dong Guo","doi":"10.1007/s11626-026-01157-z","DOIUrl":"10.1007/s11626-026-01157-z","url":null,"abstract":"<p><p>The modified Taohong Siwu decoction (MTHSWD), a traditional prescription, demonstrates notable efficacy in treating cardiovascular diseases. However, its potential mechanisms in addressing atherosclerosis (AS) require further exploration. Investigating the underlying mechanisms of MTHSWD in treating AS is imperative. AS mouse models received MTHSWD treatment, with vascular pathology and fibrosis assessed through histopathological staining. Network pharmacology identified bioactive components and potential targets, followed by molecular docking verification of key compound-target interactions. Biochemical validation included Western blot analysis. MTHSWD significantly ameliorated vascular structural abnormalities and fibrosis in AS models. Network pharmacology analysis revealed 133 active components targeting 295 proteins, intersecting with 3309 AS-related genes to form 191 overlapping targets. Twenty-two components (e.g., quercetin) demonstrated multi-target activity (≥ 15 targets). PPI network topology identified 32 hub targets including AKT1, MAPK1/8/14. Functional analysis showed that apoptosis, MAPK signaling, and lipid metabolism pathways were significantly enriched. The pathway-target network highlighted AKT1 and MAPK family members as central regulators. Molecular docking confirmed strong interactions (binding energy < -9.0 kcal/mol) between AKT1-stigmasterol/anhydroicaritin, MAPK1-emodin, and MAPK8/14-β-carotene. Experimental validation demonstrated MTHSWD's regulation of AKT1, MAPK1/14, cleaved-caspase-3, and Bcl-2 expression, enhancing endothelial viability while suppressing apoptosis. This integrated approach reveals that MTHSWD exerts anti-AS effects through multi-target modulation of PI3K/AKT and MAPK signaling pathways, promoting endothelial survival and attenuating apoptotic processes, ultimately alleviating AS-related vascular pathology.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"511-524"},"PeriodicalIF":1.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147456898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ginsenoside F1 ameliorates nonalcoholic fatty liver disease by activating the AMPK/PGC-1α pathway and autophagy.","authors":"Wenjiao Xie, Yun Lv, Zhixian Zhou, Guihui Wang, Xu Zhang, Daiping Peng, Haiyan Yang, Juan Liao, Xin Sun, Wei Lin, Zhe Ding","doi":"10.1007/s11626-026-01177-9","DOIUrl":"https://doi.org/10.1007/s11626-026-01177-9","url":null,"abstract":"<p><p>Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver diseases globally, and currently, there is a lack of specific effective drugs. Ginsenoside F1 (Gf1) is the main active ingredient of the traditional Cahinese medicine Sansheng Siwei Wan, but its role and mechanism in NAFLD remain unclear. This study aims to explore the ameliorative effect of Gf1 on NAFLD and clarify whether it exerts its effect by regulating the AMPK/PGC-1α signaling pathway and autophagy. An NAFLD model was established by feeding SD rats with a high-fat diet (HFD) for 8 weeks, followed by intragastric administration of different doses (25, 50, 100 mg/kg/d) of Gf1 for 4 weeks. In vitro, a lipotoxicity model was established by treating AML-12 mouse hepatocytes with a mixture of oleic acid/palmitic acid (OA/PA), and Gf1 (20, 40 μM) was applied for intervention. The efficacy of Gf1 was evaluated by detecting serum biochemical indicators, liver histopathology, inflammatory factors, cell proliferation, apoptosis, etc. Western Blot was used to detect the expression of key proteins in the AMPK/PGC-1α pathway and autophagy marker proteins in the liver and cells. Mechanistic verification was carried out by using the AMPK inhibitor BML-275 and the PGC-1α inhibitor SR-18292. Gf1 intervention significantly reduced the liver weight, liver index, serum levels of ALT, AST, TG, TC, and the expression of inflammatory factors TNF-α, IL-6, IFN-γ in NAFLD rats, and improved liver fat accumulation and glycogen storage. In AML-12 cells, Gf1 promoted cell proliferation inhibited by FFA, inhibited cell apoptosis and the release of inflammatory factors. Mechanistic studies showed that Gf1 significantly upregulated the protein expression of p-AMPK and PGC-1α in liver tissues and cells, and simultaneously activated autophagy (increased LC3-II/I and Beclin-1, decreased p62). The promoting effects of Gf1 on cell proliferation, apoptosis, inflammation, and autophagy could be reversed by the AMPK or PGC-1α inhibitor. Gf1 can effectively improve liver steatosis, inflammatory response, and cell damage in NAFLD models in vivo and in vitro. Its protective effect may be related to the activation of the AMPK/PGC-1α signaling pathway and the subsequent enhancement of autophagy. This study reveals for the first time the potential value of Gf1 in the treatment of NAFLD and its molecular mechanism, providing new experimental evidence for the development of NAFLD drugs targeting the AMPK/PGC-1α/autophagy axis.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147591725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Elevated intravesical pressure promotes circulating tumor cell release in an orthotopic mouse model of bladder cancer.","authors":"Ling Yi, Qing Chen, Min Zeng, Zongxin Xiao, Hua Chen, Leming Song, Xianxin Zhu","doi":"10.1007/s11626-026-01176-w","DOIUrl":"https://doi.org/10.1007/s11626-026-01176-w","url":null,"abstract":"<p><p>Elevated intravesical pressure during transurethral resection of bladder tumor (TURBT) is hypothesized to facilitate the release of circulating tumor cells (CTCs), potentially increasing the risk of hematogenous metastasis. However, the causal relationship between intravesical pressure and CTC dissemination remains unclear. This study aimed to investigate the effect of different intravesical pressure levels on the release of CTCs in an orthotopic mouse model of bladder cancer. An orthotopic bladder cancer model was established in female C57BL/6 mice by implanting MB49-luc cells. Tumor-bearing mice were randomly assigned to three groups (n = 6 per group) subjected to different intravesical pressures for 20 min: 0 cmH₂O, 40 cmH₂O, and 80 cmH₂O. Peripheral blood was collected 24 h post-intervention for CTC detection. CTCs were isolated and identified as CD45⁻/EpCAM⁺ cells using flow cytometry. The release of CTCs into the peripheral blood was significantly promoted by elevated intravesical pressure. Mice in the 40 cmH₂O and 80 cmH₂O groups showed a significantly higher proportion of CTCs compared to the 0 cmH₂O control group. Furthermore, the 80 cmH₂O group demonstrated a significantly greater increase in CTCs than the 40 cmH₂O group, indicating a pressure-dependent effect. Increased intravesical pressure directly promotes the release of CTCs in bladder cancer in a pressure-dependent manner. These findings provide experimental evidence supporting the \"pressure-driven CTC dissemination\" hypothesis and highlight the potential clinical importance of monitoring and controlling intravesical pressure during surgical procedures to minimize the risk of tumor cell spread.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147581208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"miR-3614-5p prevents endothelial differentiation and angiogenesis in human exfoliated deciduous teeth by targeting lysophosphatidic acid receptor 2.","authors":"Yiming Chen, Li Zhang, Jie Zhang, Xuezhen Peng, Ying Chen, Jing Xie","doi":"10.1007/s11626-026-01171-1","DOIUrl":"https://doi.org/10.1007/s11626-026-01171-1","url":null,"abstract":"<p><p>The contribution of microRNAs (miRNAs) to the endothelial differentiation of stem cells from human exfoliated deciduous teeth (SHED) remains largely unclear. This study sought to uncover novel miRNAs implicated in this process. SHED were obtained from deciduous teeth and verified via flow cytometric profiling of CD34, CD90, and CD105. Cells were induced toward endothelial lineage and subjected to miRNA sequencing at 0, 7, and 14 d. Differentially expressed miRNAs were validated by quantitative Polymerase Chain Reaction (qPCR). The influence of miR-3614-5p on endothelial differentiation was examined using qPCR, Western blotting, immunofluorescence, and angiogenesis assays. Direct targeting of lysophosphatidic acid receptor 2 (LPAR2) was tested through luciferase reporter assays, and rescue experiments were performed by LPAR2 overexpression. Results showed that SHED expressed CD34 (19.5%), CD90 (99.1%), and CD105 (92.5%), confirming effective isolation. Sequencing identified 9 consistently downregulated and 11 consistently upregulated miRNAs at both days 7 and 14, with miR-3614-5p denoting stable upregulation and qPCR confirmation. Functionally, miR-3614-5p exerted opposing effects: its overexpression suppressed von Willebrand factor and CD31 expression and impaired tube formation, whereas its inhibition enhanced marker expression and angiogenesis. Mechanistically, LPAR2 was validated as a direct target of miR-3614-5p. Importantly, LPAR2 overexpression reversed the inhibitory effects of miR-3614-5p on endothelial differentiation and angiogenesis. Together, these findings indicated that miR-3614-5p negatively regulates SHED endothelial differentiation and angiogenesis by directly targeting LPAR2, highlighting its potential role as a molecular regulator in this process.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147581254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanism of LPS-activated gingival fibroblasts promote the migration of gastric cancer cells via CXCL12/CXCR4-Snail2-EMT axis.","authors":"Tongbin Liu, Qi Tang, Min Liu, Fangping Ren, Rubing Liu","doi":"10.1007/s11626-026-01168-w","DOIUrl":"https://doi.org/10.1007/s11626-026-01168-w","url":null,"abstract":"<p><p>The objective of this article is to explore the influence and process by which periodontitis affects the metastasis of gastric cancer cells. Human gingival fibroblasts (HGFs) underwent stimulation with lipopolysaccharide (LPS), and ELISA was used to detect inflammatory factor expression. Using cell co-culture technology, the effect of HGFs on the migration of gastric cancer cells was observed. CCK8, qPCR, Western blot, RNAi, Cell scratch and transwell were used to analyze the relationship between CXCL12/CXCR4, Snail2 and EMT, and their effects on cancer cell migration. The results showed that CXCL12 levels rise in periodontitis and gastric cancer. HGFs can promote the migration of gastric cancer cells. CXCL12 also boosts Snail2 expression in gastric cancer cells. AMD3100 and si-CXCR4 can significantly inhibit the expression of Snail2. Moreover, CXCL12 can promote EMT, while si-Snail2 and si-CXCR4 significantly reduced the upregulation trend of N-cadherin and Vimentin expression, and reverses the downregulation trend of E-cadherin expression. CXCL12 can promote the migration of gastric cancer cells, while the migration rates of gastric cancer cells are notably reduced after AMD3100, si-CXCR4 or si-Snail2 intervention. In summary, the CXCL12/CXCR4-Salil2-EMT signaling pathway is involved in the migration of gastric cancer cells promoted by periodontitis. This will uncover new insights into the mechanisms that might play a role in the development of gastric cancer and strategies for targeted therapy.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147520872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Decorin inhibits the migration and invasion of the LPS + high glucose-induced primary trophoblast cell through ADAMTS12.","authors":"Qiuling Chen, Taoyan Hu, Hui Luo","doi":"10.1007/s11626-026-01166-y","DOIUrl":"https://doi.org/10.1007/s11626-026-01166-y","url":null,"abstract":"<p><p>Decorin was identified as the most relevant biomarker in type 2 diabetes or obesity-related diseases, but its function in gestational obesity or gestational diabetes mellitus (GDM) remains unknown. RT-qPCR or western blot were employed to validate the expression of Decorin and ADAMTS12 in the umbilical cord blood or placental tissues of the aforementioned groups. Subsequently, an LPS + high glucose-induced primary trophoblast cell model was established to verify the expression of DCN and ADAMTS12 in trophoblasts. Based on this model, silencing (si) of Decorin, or Decorin recombinant protein (DCN-r) combined with si-ADAMTS12 intervention, was performed to investigate its effects on trophoblast cell proliferation, migration, invasion, and other characteristics. Compared to the normal pregnancy group, the levels of DCN and ADAMTS12 showed an increasing trend in the gestational obesity group and the GDM group. Notably, DCN and ADAMTS12 were markedly higher in the GDM with obesity group compared to either the gestational obesity or GDM-alone groups. LPS + high glucose inhibited the migration and invasion of primary trophoblast cells while promoting increased levels of DCN and ADAMTS12. Under LPS + high-glucose intervention, DCN-r further suppressed trophoblast cell migration and invasion while enhancing ADAMTS12 expression, whereas si-DCN reversed these effects. Additionally, si-ADAMTS12 attenuated the LPS + high glucose-induced decline in trophoblast cell migration and invasion and blocked the promoting effects of DCN recombinant protein. Silencing ADAMTS12 can inhibit the promotive effect of DCN on the migration and invasion capabilities of LPS + high glucose-induced primary trophoblast cells. Clinical trial number: not applicable.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147511747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative pathophysiological modeling: the advantage of \"two-hit\" over \"one-hit\" in acute liver failure studies.","authors":"Yishu Yan, Jingping Huang, Mingzhu Chen, Liyin Li, Mengdie Lu, Jing Yang","doi":"10.1007/s11626-026-01169-9","DOIUrl":"https://doi.org/10.1007/s11626-026-01169-9","url":null,"abstract":"<p><p>Acute liver failure (ALF), a life-threatening condition marked by rapid hepatocyte death and systemic inflammation, poses significant clinical challenges due to its high mortality. The crosstalk between necrotic hepatocytes and infiltrating immune cells is hypothesized to drive disease progression. To investigate this interplay, we developed a sequential \"two-hit\" murine model using concanavalin A (Con A) challenges and compared its pathophysiological outcomes with the conventional single-dose \"one-hit\" approach. The results demonstrated that the \"two-hit\" model induced more severe hepatic coagulation dysfunction, extensive hepatocellular necrosis, destruction of liver lobular architecture, and inflammatory responses. Furthermore, serum levels of alanine transaminase (ALT) and aspartate transaminase (AST) were markedly elevated in the \"two-hit\" group. Inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were also significantly elevated. Moreover, substantial recruitment of macrophages was observed in the \"two-hit\" model, indicating that these cells are key determinants in the interaction with dying hepatocytes for the progression of ALF. Complementary ex vivo experiments revealed that Raw264.7 cells subjected to a \"two-hit\" stimulation with Con A and hepatocyte debris produced a robust inflammatory response through the classical NF-κB signaling pathway.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147490968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}