Šisl Dino, Flegar Darja, Filipović Maša, Turčić Petra, Planinić Pavao, Šućur Alan, Kovačić Nataša, Grčević Danka, Kelava Tomislav
Laboratory for Molecular Immunology, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, 10000 Zagreb, Croatia.
Department of Physiology, School of Medicine, University of Zagreb, 10000 Zagreb, Croatia.
Biomedicines. 2022 May 23;10(5):1209. doi: 10.3390/biomedicines10051209.
The available treatments for cholestatic liver fibrosis are limited, and the disease often progresses to liver cirrhosis. Tamoxifen is a selective modulator of estrogen receptors, commonly used in breast cancer therapy. A recent in vitro study showed that tamoxifen deactivates hepatic stellate cells, suggesting its potential as an antifibrotic therapeutic, but its effects in vivo remain poorly investigated. In the present study, we show that tamoxifen protects against the cholestatic fibrosis induced by a diet supplemented with 0.025% 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). Mice fed with a DDC-supplemented diet for four weeks and treated with tamoxifen developed a significantly milder degree of liver fibrosis than vehicle-treated mice, as evidenced by a lower percentage of Sirius red-stained area (60.4% decrease in stained area in male and 42% decrease in female mice, p < 0.001 and p < 0.01, respectively) and by lower hydroxyproline content. The finding was further confirmed by qPCR analysis, which showed a lower expression of genes for Col1a1, Acta2, Sox9, Pdgf, and Krt19, indicating the inhibitory effect on hepatic stellate cells, collagen production, and biliary duct proliferation. The degree of protection was similar in male and female mice. Tamoxifen per se, injected into standard-diet-fed mice, increased the expression of genes for Il6 (p < 0.01 and p < 0.001 in male and female mice, respectively) and Tgfβ (p < 0.01 for both sexes), and had no adverse effects. We showed that tamoxifen sex-independently protects against cholestatic DDC-induced liver fibrosis. The increased expression of Il6 and Tgfβ seems to be a plausible protective mechanism that should be the primary focus of further research.
目前针对胆汁淤积性肝纤维化的治疗方法有限,且该疾病常进展为肝硬化。他莫昔芬是一种雌激素受体的选择性调节剂,常用于乳腺癌治疗。最近的一项体外研究表明,他莫昔芬可使肝星状细胞失活,提示其具有抗纤维化治疗的潜力,但对其体内作用的研究仍较少。在本研究中,我们发现他莫昔芬可预防由添加0.025% 3,5 - 二乙氧基羰基 - 1,4 - 二氢可力丁(DDC)的饮食诱导的胆汁淤积性纤维化。喂食含DDC饮食四周并接受他莫昔芬治疗的小鼠,其肝纤维化程度明显轻于接受载体治疗的小鼠,这通过较低的天狼星红染色面积百分比(雄性小鼠染色面积减少60.4%,雌性小鼠减少42%,p分别<0.001和p<0.01)以及较低的羟脯氨酸含量得以证明。qPCR分析进一步证实了这一发现,该分析显示Col1a1、Acta2、Sox9、Pdgf和Krt19基因的表达较低,表明对肝星状细胞、胶原蛋白生成和胆管增殖具有抑制作用。雄性和雌性小鼠的保护程度相似。将他莫昔芬注射到喂食标准饮食的小鼠体内,本身会增加Il6基因(雄性和雌性小鼠分别为p<0.01和p<0.001)和Tgfβ基因(两性均为p<0.01)的表达,且无不良影响。我们表明,他莫昔芬可不分性别地预防胆汁淤积性DDC诱导的肝纤维化。Il6和Tgfβ表达的增加似乎是一种合理的保护机制,应成为进一步研究的主要重点。