Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan.
Department of Biochemistry, Faculty of Pharmacy, Minia University, Minia 61519, Egypt.
Molecules. 2020 Apr 26;25(9):2020. doi: 10.3390/molecules25092020.
Mesenchymal stem cells (MSCs) have considerable therapeutic abilities in various disorders, including hepatic fibrosis. They may be affected with different culture conditions. This study investigated, on molecular basics, the effect of pretreatment with eugenol on the characteristics of adipose tissue-derived MSCs (ASCs) in vitro and the implication of eugenol preconditioning on the in vivo therapeutic abilities of ASCs against CCl-induced hepatic fibrosis in rats. The effect of eugenol on ASCs was assessed using viability, scratch migration and sphere formation assays. Expressions of genes and proteins were estimated by immunofluorescence or qRT-PCR. For the in vivo investigations, rats were divided into four groups: the normal control group, fibrotic (CCl) group, CCl+ASCs group and CCl + eugenol-preconditioned ASCs (CCl+E-ASCs) group. Eugenol affected the viability of ASCs in a concentration- and time-dependent manner. Eugenol improved their self-renewal, proliferation and migration abilities and significantly increased their expression of , reduced expression 1 (), octamer-binding transcription factor 4 () and genes. Furthermore, E-ASCs showed more of a homing ability than ASCs and improved the serum levels of ALT, AST, albumin, total bilirubin and hyaluronic acid more efficient than ASCs in treating CCl-induced hepatic fibrosis, which was confirmed with histopathology. More interestingly, compared to the CCl+ASCs group, CCl+E-ASCs group showed a lower expression of inducible nitric oxide synthase (), monocyte chemoattractant protein-1 (), cluster of differentiation 163 () and tumor necrosis factor-α () genes and higher expression of matrix metalloproteinase ()-9 and genes. This study, for the first time, revealed that eugenol significantly improved the self-renewal, migration and proliferation characteristics of ASCs, in vitro. In addition, we demonstrated that eugenol-preconditioning significantly enhanced the therapeutic abilities of the injected ASCs against CCl-induced hepatic fibrosis.
间充质干细胞 (MSCs) 在多种疾病中具有相当大的治疗能力，包括肝纤维化。它们可能会受到不同培养条件的影响。本研究从分子基础上探讨了丁香酚预处理对脂肪组织来源的间充质干细胞 (ASCs) 体外特性的影响，以及丁香酚预处理对 ASCs 治疗 CCl 诱导的大鼠肝纤维化的体内治疗能力的影响。使用细胞活力、划痕迁移和球体形成试验评估丁香酚对 ASCs 的影响。通过免疫荧光或 qRT-PCR 评估基因和蛋白质的表达。对于体内研究，将大鼠分为四组：正常对照组、纤维化组 (CCl 组)、CCl+ASCs 组和 CCl+丁香酚预处理 ASCs (CCl+E-ASCs) 组。丁香酚以浓度和时间依赖的方式影响 ASCs 的活力。丁香酚改善了它们的自我更新、增殖和迁移能力，并显著增加了它们的 、下调了 1 (), 八聚体结合转录因子 4 () 和 基因的表达。此外，E-ASCs 比 ASCs 具有更强的归巢能力，并在治疗 CCl 诱导的肝纤维化方面比 ASCs 更有效地提高了血清 ALT、AST、白蛋白、总胆红素和透明质酸水平，这在组织病理学上得到了证实。更有趣的是，与 CCl+ASCs 组相比，CCl+E-ASCs 组的诱导型一氧化氮合酶 ()、单核细胞趋化蛋白-1 (), 分化簇 163 () 和肿瘤坏死因子-α () 基因的表达降低，基质金属蛋白酶 ()-9 和 基因的表达升高。本研究首次揭示，丁香酚显著改善了 ASCs 的自我更新、迁移和增殖特性，体外。此外，我们证明了丁香酚预处理显著增强了注射 ASCs 对 CCl 诱导的肝纤维化的治疗能力。
