Fatima Abiha, Malick Tuba Shakil, Khan Irfan, Ishaque Aisha, Salim Asmat
Stem Cell Laboratory, Dr. Panjwani Center for Molecular Medicine and Drug Research, ICCBS, University of Karachi, Karachi 75270, Sindh, Pakistan.
World J Stem Cells. 2021 Oct 26;13(10):1580-1594. doi: 10.4252/wjsc.v13.i10.1580.
End-stage liver disease is a global health complication with high prevalence and limited treatment options. Cell-based therapies using mesenchymal stem cells (MSCs) emerged as an alternative approach to support hepatic regeneration. preconditioning strategies have been employed to strengthen the regenerative and differentiation potential of MSCs towards hepatic lineage. Chemical compounds of the triterpene class; glycyrrhizic acid (GA) and 18β-glycyrrhetinic acid (GT) possess diverse therapeutic properties including hepato-protection and anti-fibrosis characteristics. They are capable of modulating several signaling pathways that are crucial in hepatic regeneration. Preconditioning with hepato-protective triterpenes may stimulate MSC fate transition towards hepatocytes.
To explore the effect of GA and GT on hepatic differentiation of human umbilical cord-MSCs (hUC-MSCs).
hUC-MSCs were isolated and characterized phenotypically by flow cytometry and immunocytochemistry for the expression of MSC-associated surface molecules. Isolated cells were treated with GA, GT, and their combination for 24 h and then analyzed at three time points; day 7, 14, and 21. qRT-PCR was performed for the expression of hepatic genes. Expression of hepatic proteins was analyzed by immunocytochemistry at day 21. Periodic acid Schiff staining was performed to determine the functional ability of treated cells.
The fusiform-shaped morphology of MSCs in the treatment groups in comparison with the untreated control, eventually progressed towards the polygonal morphology of hepatocytes with the passage of time. The temporal transcriptional profile of preconditioned MSCs displayed significant expression of hepatic genes with increasing time of differentiation. Preconditioned cells showed positive expression of hepatocyte-specific proteins. The results were further corroborated by positive periodic acid Schiff staining, indicating the presence of glycogen in their cytoplasm. Moreover, bi-nucleated cells, which is the typical feature of hepatocytes, were also seen in the preconditioned cells.
Preconditioning with glycyrrhizic acid, 18β-glycyrrhetinic acid and their combination, successfully differentiates hUC-MSCs into hepatic-like cells. These MSCs may serve as a better therapeutic option for degenerative liver diseases in future.
终末期肝病是一种全球范围内普遍存在且治疗选择有限的健康并发症。使用间充质干细胞（MSCs）的细胞疗法成为支持肝脏再生的一种替代方法。预处理策略已被用于增强MSCs向肝系细胞的再生和分化潜能。三萜类化合物甘草酸（GA）和18β-甘草次酸（GT）具有多种治疗特性，包括肝脏保护和抗纤维化特性。它们能够调节肝脏再生中至关重要的几种信号通路。用具有肝脏保护作用的三萜类化合物进行预处理可能会刺激MSCs向肝细胞的命运转变。
探讨GA和GT对人脐带间充质干细胞（hUC-MSCs）肝向分化的影响。
分离hUC-MSCs，并通过流式细胞术和免疫细胞化学对其进行表型鉴定，以检测MSCs相关表面分子的表达。将分离的细胞用GA、GT及其组合处理24小时，然后在第7天、14天和21天这三个时间点进行分析。通过qRT-PCR检测肝脏基因的表达。在第21天通过免疫细胞化学分析肝脏蛋白的表达。进行过碘酸希夫染色以确定处理后细胞的功能能力。
与未处理的对照组相比，处理组中MSCs的梭形形态最终随着时间的推移发展为肝细胞的多边形形态。预处理后的MSCs的时间转录谱显示，随着分化时间的增加，肝脏基因有显著表达。预处理后的细胞显示出肝细胞特异性蛋白的阳性表达。过碘酸希夫染色阳性进一步证实了该结果，表明其细胞质中存在糖原。此外，在预处理后的细胞中还观察到了双核细胞，这是肝细胞的典型特征。
用甘草酸、18β-甘草次酸及其组合进行预处理可成功地将hUC-MSCs分化为肝样细胞。这些MSCs未来可能成为退行性肝病更好的治疗选择。
