Kartasheva-Ebertz Daria, Gaston Jesintha, Lair-Mehiri Loriane, Massault Pierre-Philippe, Scatton Olivier, Vaillant Jean-Christophe, Morozov Vladimir Alexei, Pol Stanislas, Lagaye Sylvie
Institut Pasteur, Immunobiologie des Cellules Dendritiques, INSERM U1223, Paris 75015, France.
Service de Chirurgie digestive, Hépato-biliaire et Endocrinienne, AP-HP, Groupe Hospitalier Cochin, Paris 75014, France.
World J Hepatol. 2021 Feb 27;13(2):187-217. doi: 10.4254/wjh.v13.i2.187.
Liver fibrosis can result in end-stage liver failure and death.
To examine human liver fibrogenesis and anti-fibrotic therapies, we evaluated the three dimensional liver slice (LS) model.
Fibrotic liver samples (F0 to F4 fibrosis stage according to the METAVIR score) were collected from patients after liver resection. Human liver slices (HLS) were cultivated for up to 21 days. Hepatitis C virus (HCV) infection, alcohol (ethanol stimulation) and steatosis (palmitate stimulation) were examined in fibrotic (F2 to F4) liver slices infected (or not) with HCV. F0-F1 HLS were used as controls. At day 0, either ursodeoxycholic acid (choleretic and hepatoprotective properties) and/or α-tocopherol (antioxidant properties) were added to standard of care on HLS and fibrotic liver slices, infected (or not) with HCV. Expression of the biomarkers of fibrosis and the triglyceride production were checked by quantitative reverse transcription polymerase chain reaction and/or enzyme-linked immunosorbent assay.
The cultures were viable for 21 days allowing to study fibrosis inducers and to estimate the effect of anti-fibrotic drugs. Expression of the biomarkers of fibrosis and the progression to steatosis (estimated by triglycerides production) was increased with the addition of HCV and /or ethanol or palmitate. From day 15 of the follow-up studies, a significant decrease of both transforming growth factor β-1 and Procol1A1 expression and triglycerides production was observed when a combined anti-fibrotic treatment was applied on HCV infected F2-F4 LS cultures.
These results show that the human three dimensional model effectively reflects the processes in damaged human liver (viral, alcoholic, nonalcoholic steatohepatitis liver diseases) and provides the proof of concept that the LS examined model permits a rapid evaluation of new anti-fibrotic therapies when used alone or in combination.
肝纤维化可导致终末期肝衰竭和死亡。
为研究人类肝纤维化形成及抗纤维化治疗，我们评估了三维肝切片（LS）模型。
肝切除术后从患者处收集纤维化肝样本（根据METAVIR评分分为F0至F4纤维化阶段）。将人类肝切片（HLS）培养长达21天。在感染（或未感染）丙型肝炎病毒（HCV）的纤维化（F2至F4）肝切片中检测HCV感染、酒精（乙醇刺激）和脂肪变性（棕榈酸刺激）情况。F0 - F1 HLS用作对照。在第0天，将熊去氧胆酸（具有利胆和肝保护特性）和/或α - 生育酚（具有抗氧化特性）添加到HLS和感染（或未感染）HCV的纤维化肝切片的标准护理中。通过定量逆转录聚合酶链反应和/或酶联免疫吸附测定检查纤维化生物标志物的表达和甘油三酯生成情况。
培养物在21天内保持活力，从而能够研究纤维化诱导因素并评估抗纤维化药物的效果。添加HCV和/或乙醇或棕榈酸后，纤维化生物标志物的表达以及向脂肪变性的进展（通过甘油三酯生成评估）增加。在随访研究的第15天起，当对感染HCV的F2 - F4 LS培养物应用联合抗纤维化治疗时，观察到转化生长因子β - 1和Procol1A1表达以及甘油三酯生成均显著降低。
这些结果表明，人类三维模型有效地反映了受损人类肝脏（病毒性、酒精性、非酒精性脂肪性肝炎肝病）中的过程，并提供了概念验证，即所检测的LS模型在单独使用或联合使用时允许快速评估新的抗纤维化疗法。
