Quelhas Patrícia, Oliveira Rui, Kieling Carlos, Vieira Sandra, Dos Santos Jorge
Faculty of Health Sciences, Health Science Investigation Center of University of Beira Interior (CICS-UBI), 6200-506 Covilhã, Portugal.
Coimbra Institute for Clinical and Biomedical Research (iCBR), Area of Environment, Genetics and Oncobiology (CIMAGO), Faculty of Medicine, University of Coimbra, 3000-548 Coimbra, Portugal.
Biomedicines. 2025 Jan 1;13(1):87. doi: 10.3390/biomedicines13010087.
Biliary atresia (BA) is a progressive hepatobiliary disease in infants, leading to liver failure and the need for transplantation. While its etiopathogenesis remains unclear, recent studies suggest primary cilia (PC) disruption plays a role. This study investigates correlations between PC and cytoplasmic tubulin (TUBA4A) alterations with hypoxia in patients with the isolated form of BA, focusing on native liver survival. Using qualitative and quantitative digital image analysis of immunofluorescence-stained liver samples, we assessed PC and TUBA4A features correlating these findings with HIF-1α nuclear positivity, clinical-laboratory data, and early native liver survival. Liver samples from fourteen BA patients and six controls with another liver disease were analyzed by digital image analysis, with data evaluated using Spearman's correlation and independent -tests. HIF-1α positivity in cholangiocytes was observed in 42.8% of BA patients. While the PC ratio per biliary structure (cilia ratio status, CRs) was similar between BA patients and controls, PC length was decreased in BA patients. Cytoplasmic TUBA4A levels were elevated in BA patients. CRs positively correlated with lower cytoplasmic TUBA4A expression and was higher in patients without HIF-1α nuclear positivity. Reduced cilia length correlated with higher bilirubin levels at portoenterostomy. Predictors of early poor prognosis (death or need for transplantation until 1 year of life) included HIF-1α positivity, elevated direct bilirubin levels, decreased cilia length, PC bending, and increased TUBA4A expression. Reduced PC length, PC bending, and increased intensity of cytoplasmic TUBA4A expression occur in the isolated BA clinical type and negatively impact the early prognosis after post-portoenterostomy. These findings suggest the existence of a disruption in the tubulin transport between cytoplasm and PC. The detrimental effect of HIF-1alpha pathway activation over early native liver survival was confirmed, although independently from PC or cytoplasmic tubulin features.
胆道闭锁（BA）是一种发生于婴儿的进行性肝胆疾病，可导致肝衰竭并需要进行移植。虽然其病因发病机制尚不清楚，但最近的研究表明原发性纤毛（PC）破坏起了一定作用。本研究调查孤立型BA患者中PC和细胞质微管蛋白（TUBA4A）改变与缺氧之间的相关性，重点关注自体肝存活率。通过对免疫荧光染色的肝脏样本进行定性和定量数字图像分析，我们评估了PC和TUBA4A特征，并将这些发现与缺氧诱导因子-1α（HIF-1α）核阳性、临床实验室数据以及早期自体肝存活率相关联。通过数字图像分析对14例BA患者和6例患有其他肝脏疾病的对照者的肝脏样本进行分析，并使用Spearman相关性分析和独立检验对数据进行评估。在42.8%的BA患者中观察到胆管细胞中HIF-1α呈阳性。虽然BA患者和对照者之间每个胆管结构的PC比率（纤毛比率状态，CRs）相似，但BA患者的PC长度缩短。BA患者的细胞质TUBA4A水平升高。CRs与较低的细胞质TUBA4A表达呈正相关，且在无HIF-1α核阳性的患者中更高。纤毛长度缩短与门肠吻合术时较高的胆红素水平相关。早期预后不良（死亡或在1岁前需要移植）的预测因素包括HIF-1α阳性、直接胆红素水平升高、纤毛长度缩短、PC弯曲以及TUBA4A表达增加。孤立型BA临床类型中出现PC长度缩短、PC弯曲以及细胞质TUBA4A表达强度增加，对门肠吻合术后的早期预后产生负面影响。这些发现表明细胞质与PC之间的微管蛋白转运存在破坏。尽管独立于PC或细胞质微管蛋白特征，但证实了HIF-1α通路激活对早期自体肝存活率有不利影响。
