Amer Johnny, Grifat Rami, Doron Sarit, Abu-Tair Lina, Mruwat Rufayda, El-Khatib Areej, Safadi Rifaat
The Liver and Gastroenterology Units, Division of Medicine, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
Liver Int. 2014 Sep;34(8):1232-40. doi: 10.1111/liv.12371. Epub 2013 Dec 10.
BACKGROUND & AIMS: Immune cells interact with hepatic-stellate-cells (HSCs) in the development of liver fibrosis. Little is known about the influence of pregnancy on the development and progression of hepatic-fibrosis. In this study, we explored the influence of pregnancy on progression of hepatic fibrosis.
Female mice (C57Blc) were induced by 4 injections of peritoneal carbon-tetrachloride (CCl4) within 10 days, starting at day 10 of documented pregnancy. At end of experiment, serum samples were obtained for ALT and estradiol determination. Harvested livers were histological evaluated for liver injury and for protein αSMA expressions. Isolated intra-hepatic lymphocytes were assessed by flow cytometry. Isolated lymphocytes and serum samples were in- vitro co-cultured for 48 h with primary isolated naïve HSCs. Washed cells were analyzed for adherence (anti-αSMA+/anti-CD45 + ) and proliferations (CSFE).
CCl4-model for liver injury was well tolerated when induced in pregnancy similar to non-pregnant state. Hepatic-fibrosis (Masson Trichrome Stain, Sirius red stain and αSMA expressions) and necro-inflammation (H&E stain and serum ALT levels) significantly increased in pregnancy. Increased liver injury was accompanied with pro-fibrotic lymphocyte profile; CD8 subsets increased and NK cells decreased. HSCs activation significantly increased when in-vitro cultured with lymphocytes from pregnant as compared to non-pregnant fibrotic ones. Pro-fibrotic profile was also explained by decreased NK activity (CD107a marker) and of their phagocytosis. Serum estradiol levels although elevated in fibrosis conditions of pregnancy was not associated with the pHSCs activations.
Liver fibrosis in our murine model was severe in pregnant model; via pro-fibrotic lymphocyte and serum alterations.
在肝纤维化发展过程中，免疫细胞与肝星状细胞（HSCs）相互作用。关于妊娠对肝纤维化发展和进程的影响知之甚少。在本研究中，我们探讨了妊娠对肝纤维化进程的影响。
从记录的妊娠第10天开始，在10天内对雌性小鼠（C57Blc）进行4次腹腔注射四氯化碳（CCl4）诱导。实验结束时，采集血清样本测定谷丙转氨酶（ALT）和雌二醇。对收获的肝脏进行组织学评估，以检测肝损伤和α平滑肌肌动蛋白（αSMA）蛋白表达。通过流式细胞术评估分离的肝内淋巴细胞。将分离的淋巴细胞和血清样本与原代分离的未活化HSCs进行48小时的体外共培养。对洗涤后的细胞进行黏附（抗αSMA + /抗CD45 +）和增殖（羧基荧光素二乙酸琥珀酰亚胺酯，CSFE）分析。
与非妊娠状态相似，在妊娠期间诱导的CCl4肝损伤模型耐受性良好。妊娠期间肝纤维化（Masson三色染色、天狼星红染色和αSMA表达）和坏死性炎症（苏木精-伊红染色和血清ALT水平）显著增加。肝损伤增加伴随着促纤维化淋巴细胞谱；CD8亚群增加，自然杀伤（NK）细胞减少。与来自非妊娠纤维化小鼠的淋巴细胞相比，与来自妊娠小鼠的淋巴细胞进行体外培养时，HSCs活化显著增加。促纤维化谱也可通过NK活性（CD107a标志物）及其吞噬作用的降低来解释。尽管妊娠纤维化状态下血清雌二醇水平升高，但与pHSCs活化无关。
在我们的小鼠模型中，妊娠模型中的肝纤维化严重；通过促纤维化淋巴细胞和血清改变。
