Mortezaee Keywan, Pasbakhsh Parichehr, Ragerdi Kashani Iraj, Sabbaghziarani Fatemeh, Omidi Ameneh, Zendedel Adib, Ghasemi Soudabeh, Dehpour Ahmad Reza
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Institute of Neuroanatomy, School of Medicine, RWTH Aachen University, 52074 Aachen, Germany.
Iran Biomed J. 2016 Sep;20(4):207-16. doi: 10.7508/ibj.2016.04.004. Epub 2016 Apr 30.
Bone marrow-derived mesenchymal stem cells (BMMSCs) transplantation has been considered as a promising milestone in liver fibrosis treatment. However, low amounts of homing are a major obstacle. We aimed to investigate the role of melatonin pretreatment in BMMSC homing into experimental liver fibrosis.
BMMSCs were obtained, grown, propagated and preconditioned with 5 µM melatonin and analyzed for multipotency and immunophenotypic features at passage three. The cells were labelled with CM-Dil and infused into the rats received the i.p. injection of carbon tetrachloride (CCl4) for five weeks to induce liver fibrosis. Animals were divided into two groups: One group received BMMSCs, whereas the other group received melatonin-pretreated BMMSCs (MT-BMMSCs). After cell injection at 72 h, animals were sacrificed, and the liver tissues were assessed for further evaluations: fibrosis using Masson's trichrome and hematoxylin and eosin staining and homing using fluorescent microscopy and flow cytometry.
BMMSCs and MT-BMMSCs expressed a high level of CD44 but low levels of CD11b, CD45 and CD34 (for all P≤0.05) and were able to differentiate into adipocytes and Schwann cells. CCl4 induction resulted in extensive collagen deposition, tissue disruption and fatty accumulation with no obvious difference between the two groups. There was a significant increase in homing of MT-BMMSCs in both florescent microscopy (P≤0.001) and flow cytometry (P≤0.01) assays, as compared with non-treated BMMSCs.
This study indicates the improved homing potential of BMMSCs in pretreatment with melatonin. Therefore, this strategy may represent an applied approach for improving the stem cell therapy of liver fibrosis.
骨髓间充质干细胞(BMMSCs)移植被认为是肝纤维化治疗中一个有前景的里程碑。然而,归巢数量少是一个主要障碍。我们旨在研究褪黑素预处理在BMMSCs归巢至实验性肝纤维化中的作用。
获取BMMSCs,进行培养、传代并用5 μM褪黑素预处理,在第三代分析其多能性和免疫表型特征。细胞用CM-Dil标记,注入腹腔注射四氯化碳(CCl4)5周以诱导肝纤维化的大鼠体内。动物分为两组:一组接受BMMSCs,另一组接受褪黑素预处理的BMMSCs(MT-BMMSCs)。细胞注射72小时后,处死动物,评估肝脏组织以进行进一步评估:使用Masson三色染色和苏木精-伊红染色评估纤维化,使用荧光显微镜和流式细胞术评估归巢情况。
BMMSCs和MT-BMMSCs均高表达CD44,但低表达CD11b、CD45和CD34(所有P≤0.05),并且能够分化为脂肪细胞和雪旺细胞。CCl4诱导导致广泛的胶原沉积、组织破坏和脂肪堆积,两组之间无明显差异。与未处理的BMMSCs相比,在荧光显微镜(P≤0.001)和流式细胞术(P≤0.01)分析中,MT-BMMSCs的归巢均显著增加。
本研究表明褪黑素预处理可提高BMMSCs的归巢潜力。因此,该策略可能代表一种改善肝纤维化干细胞治疗的应用方法。