Ling Li, Shu Han, Huang Yubin, Hou Jiying, Hua Yuanyuan
Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Department of Histology and Embryology, Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqing 400010, China.
Stem Cells Int. 2023 Feb 15;2023:7074703. doi: 10.1155/2023/7074703. eCollection 2023.
Ginsenoside Rg1 (Rg1) is purified from ginseng with various pharmacological effects, which might facilitate the biological behavior of human amnion-derived mesenchymal stem/stromal cells (hAD-MSCs). This study is aimed at investigating the effects of Rg1 on the biological behavior, such as viability, proliferation, apoptosis, senescence, migration, and paracrine, of hAD-MSCs. hAD-MSCs were isolated from human amnions. The effects of Rg1 on the viability, proliferation, apoptosis, senescence, migration, and paracrine of hAD-MSCs were detected by CCK-8, EdU, flow cytometry, SA--Gal staining, wound healing, and ELISA assays, respectively. The protein expression levels were detected by western blot. Cell cycle distribution was evaluated using flow cytometry. We found that Rg1 promoted hAD-MSC cycle progression from G0/G1 to S and G2/M phases and significantly increased hAD-MSC proliferation rate. Rg1 activated PI3K/AKT signaling pathway and significantly upregulated the expressions of cyclin D, cyclin E, CDK4, and CDK2 in hAD-MSCs. Inhibition of PI3K/AKT signaling significantly downregulated the expressions of cyclin D, cyclin E, CDK4, and CDK2, prevented cell cycle progression, and reduced hAD-MSC proliferation induced by Rg1. hAD-MSC senescence rate was significantly increased by D-galactose, while the elevated hAD-MSC senescence rate induced by D-galactose was significantly decreased by Rg1 treatment. D-galactose significantly induced the expressions of senescence markers, p16, p14, p21, and p53 in hAD-MSCs, while Rg1 significantly reduced the expressions of those markers induced by D-galactose in hAD-MSCs. Rg1 significantly promoted the secretion of IGF-I in hAD-MSCs. Rg1 reduced the hAD-MSC apoptosis rate. However, the difference was not significant. Rg1 had no influence on hAD-MSC migration. Altogether, our results demonstrate that Rg1 can promote the viability, proliferation, and paracrine and relieve the senescence of hAD-MSCs. PI3K/AKT signaling pathway is involved in the promotive effect of Rg1 on hAD-MSC proliferation. The protective effect of Rg1 on hAD-MSC senescence may be achieved via the downregulation of p16 and p53/p21 pathway.
人参皂苷Rg1（Rg1）是从人参中提纯的，具有多种药理作用，可能会促进人羊膜间充质干/基质细胞（hAD-MSCs）的生物学行为。本研究旨在探讨Rg1对hAD-MSCs生物学行为的影响，如活力、增殖、凋亡、衰老、迁移和旁分泌。hAD-MSCs从人羊膜中分离得到。分别通过CCK-8、EdU、流式细胞术、SA-β-Gal染色、伤口愈合和ELISA检测Rg1对hAD-MSCs活力、增殖、凋亡、衰老、迁移和旁分泌的影响。通过蛋白质印迹法检测蛋白表达水平。使用流式细胞术评估细胞周期分布。我们发现Rg1促进hAD-MSC细胞周期从G0/G1期进展到S期和G2/M期，并显著提高hAD-MSCs的增殖率。Rg1激活PI3K/AKT信号通路，并显著上调hAD-MSCs中细胞周期蛋白D、细胞周期蛋白E、细胞周期蛋白依赖性激酶4（CDK4）和细胞周期蛋白依赖性激酶2（CDK2）的表达。抑制PI3K/AKT信号通路显著下调细胞周期蛋白D、细胞周期蛋白E、CDK4和CDK2的表达，阻止细胞周期进展，并降低Rg1诱导的hAD-MSCs增殖。D-半乳糖显著提高hAD-MSCs的衰老率，而Rg1处理显著降低D-半乳糖诱导的hAD-MSCs衰老率升高。D-半乳糖显著诱导hAD-MSCs中衰老标志物p16、p14、p21和p53的表达，而Rg1显著降低D-半乳糖在hAD-MSCs中诱导的这些标志物的表达。Rg1显著促进hAD-MSCs中胰岛素样生长因子-I（IGF-I）的分泌。Rg1降低hAD-MSCs的凋亡率。然而，差异不显著。Rg1对hAD-MSCs迁移没有影响。总之，我们的结果表明Rg1可以促进hAD-MSCs的活力、增殖和旁分泌，并缓解其衰老。PI3K/AKT信号通路参与Rg1对hAD-MSCs增殖的促进作用。Rg1对hAD-MSCs衰老保护作用可能是通过下调p16和p53/p21通路实现的。
