草药预处理诱导华通氏胶间充质干细胞增殖并延缓衰老。

Herbal pre-conditioning induces proliferation and delays senescence in Wharton's Jelly Mesenchymal Stem Cells.

机构信息

Department of Biotechnology, Sinhgad College of Engineering, Affiliated to Savitribai Phule Pune University, Pune 411041, India.

School of Regenerative Medicine, Manipal University, Bangalore 560065, India.

出版信息

Biomed Pharmacother. 2017 Sep;93:772-778. doi: 10.1016/j.biopha.2017.06.107. Epub 2017 Jul 15.

DOI:10.1016/j.biopha.2017.06.107

PMID:28724259

Abstract

BACKGROUND

Mesenchymal Stem Cells (MSCs) are multipotent stem cells which are being explored for various clinical applications. Isolation and in-vitro expansion of MSCs remain important in achieving desired cell number for the therapy. However, in-vitro proliferation of MSCs is often associated with senescence and early onset of apoptosis which limits its therapeutic ability and long term clinical use. Tinospora cordifolia and Withania somnifera are used widely in Ayurveda: the traditional Indian system of medicine and are reported to have rejuvenating and anti-aging potential. In the present study, we investigated the effect of Tinospora cordifolia and Withania somnifera on proliferation and senescence of wharton's jelly MSCs (WJMSCs) in-vitro.

METHODS

WJMSCs were treated in culture medium with Tinospora cordifolia leaf and Withania somnifera root extracts to examine their effect on proliferation and senescence properties of WJMSCs. Proliferation of WJMSCs was assayed by cell count, MTT, BrdU incorporation assay, cell cycle analysis and Ki67 mRNA expression. Senescence was demonstrated using β-galactosidase senescence assay and associated mRNA markers.

RESULTS

Culture medium supplemented with Tinospora cordifolia leaf and Withania somnifera root extracts exhibited significant increase in proliferation of WJMSCs as evidenced by cell count and MTT assay. Cell cycle analysis using propidium iodide showed increase in G2/M phase and decrease in apoptotic cells. BrdU incorporation and upregulation of proliferation marker ki67 by RT PCR showed increased DNA synthesis/proliferation in Tinospora cordifolia and Withania somnifera extract treated MSCs. Delayed senescence was confirmed by β-galactosidase senescence assay and down regulation of senescence marker p21.

CONCLUSION

Our results demonstrate for the first time that Tinospora cordifolia and Withania somnifera extracts support proliferation and inhibit senescence in WJMSCs making them suitable candidates as supplements for in-vitro expansion without affecting the cell viability indicating its non-toxic nature.

摘要

背景

间充质干细胞（MSCs）是多能干细胞，正在探索用于各种临床应用。MSCs 的分离和体外扩增对于实现治疗所需的细胞数量仍然很重要。然而，MSCs 的体外增殖常伴有衰老和早期凋亡，这限制了其治疗能力和长期临床应用。三叶鬼针草和睡茄在阿育吠陀中被广泛使用：印度传统医学体系，并被报道具有 rejuvenating 和抗衰老的潜力。在本研究中，我们研究了三叶鬼针草和睡茄对体外 Wharton 胶 MSC（WJMSCs）增殖和衰老的影响。

方法

在含有三叶鬼针草叶和睡茄根提取物的培养基中处理 WJMSCs，以检测其对 WJMSCs 增殖和衰老特性的影响。通过细胞计数、MTT、BrdU 掺入测定、细胞周期分析和 Ki67 mRNA 表达来检测 WJMSCs 的增殖。通过β-半乳糖苷酶衰老测定和相关的 mRNA 标志物来证明衰老。

结果

补充有三叶鬼针草叶和睡茄根提取物的培养基显著增加了 WJMSCs 的增殖，这可通过细胞计数和 MTT 测定来证明。碘化丙啶的细胞周期分析显示 G2/M 期增加，凋亡细胞减少。BrdU 掺入和 RT-PCR 上调增殖标志物 ki67 表明 Tinospora cordifolia 和 Withania somnifera 提取物处理的 MSC 中 DNA 合成/增殖增加。β-半乳糖苷酶衰老测定和衰老标志物 p21 的下调证实了衰老的延迟。