水飞蓟素抵抗血清撤除的脐带间充质干细胞凋亡

Silymarin Protects Umbilical Cord-Derived Mesenchymal Stem Cells against Apoptosis Induced by Serum-Deprivation

目的:探讨水飞蓟素抵抗血清引起的人间充质干细胞(MSC)凋亡及其相关机制。方法:培养人脐带MSC体系去血清,加入不同浓度(1-10μg/ml)的水飞蓟素,应用MTT实验检测细胞增殖状态。细胞培养72 h后,Annexin V-PI双标记流式细胞术观察处理组和对照组细胞凋亡情况。收集细胞,以未去血清组为阳性对照,Western blot分析细胞内凋亡蛋白BCL-2和BAX含量。结果:水飞蓟素可以促进脐带MSC的增殖,效应呈浓度依赖性,浓度为5μg/ml时其促增殖作用最为明显。水飞蓟素可抑制由血清撤除引起的MSC凋亡,浓度为5μg/ml时抑制率可达30%左右。血清撤除后,细胞内BAX蛋白含量明显上升,而BCL-2含量无显著变化。结论:水飞蓟素可促进脐带间充质干细胞的增殖,并可能通过线粒体途径抑制MSC凋亡。

Objective: To investigate the protection of silymarin against the human mesenchymal stem cell( MSC)apoptosis induced by serum deprivation and its underlying mechanism. Methods: Human umbilical cord MSCs were cultured in the absence of serum,and the silymain of different concentration( 1- 10 μg /ml) was added into the medium. MTT test was performed to observe the cell proliferation status. After being cultured for 72 hours,the cells were collected,and flow cytometry with Annexin-V-PI double-staining was used to detect the apoptotic cells from the control and silymarin-treated groups. Furthermore,the intracellular contents of BAX and BCL-2 were detected by Western blot for exploring the potential mechanism. Results: The silymarin promoted the proliferation of human UCMSCs in a dose-dependent manner,reaching its maximal at a dose of 5 μg /ml. Moreover,silymarin could inhibit the serum deprivation- induced apoptosis of MSCs and,the inhibitory rate reached up to 30% when it was added at a concentration of 5 μg /ml. The content of intracellular BAX was obviously elevated after serum-deprivation treatment,and this increase could be blunted by the addition of silymarin. Meanwhile,the content of BCL-2 was not obviously changed. Conclusion: The silymarin can stimulate MSC growth and inhibit the apoptosis of MSCs probably by the mitochondria pathway.