Flavored Guilu Erxian decoction inhibits the injury of human bone marrow mesenchymal stem cells induced by cisplatin
Flavored Guilu Erxian decoction inhibits the injury of human bone marrow mesenchymal stem cells induced by cisplatin
复制标题
加味桂鹿二仙汤抑制顺铂诱导的人骨髓间充质干细胞损伤
DOI：
10.14715/cmb/2018.64.6.11
复制
发表时间：
2018-01-01
影响因子：
1.6
通讯作者：
Wang, Xilong
中科院分区：
文献类型：
--
作者：
Ke, Bin;Shi, Lin;Wang, Xilong
目的：探讨加味龟鹿二仙汤对顺铂诱导骨髓间充质干细胞（BM-MSCs）毒性反应的治疗作用。取SD大鼠骨髓分离BM-MSCs，流式细胞仪鉴定。将细胞培养在含有5%（TCM-L）、10%（TCM-M）和20%（TCM-H）剂量的加味龟鹿二仙汤（有或没有顺铂）的MEM α培养基中。通过CCK-8和胸苷类似物5-乙炔基-2 '-脱氧尿苷（EdU）染色测定来确定细胞活力。流式细胞仪检测细胞周期和凋亡。Western blot检测p21和c-caspase-3蛋白的表达。Western blot检测PI 3 K-AKT-mTOR通路相关蛋白p-PI 3 K、p-AKT和p-mTOR。CCK-8和EdU染色结果显示，顺铂可抑制BM-MSCs的增殖，且呈剂量和时间依赖性。顺铂可通过增加G 0/GI期细胞阻滞、p21和cleaved-caspase-3的表达诱导细胞凋亡。加入加味龟鹿二仙汤含药血清后，上述现象明显减轻。顺铂可抑制BM-MSCs中PI 3 K-AKT-mTOR通路的激活，而加味龟鹿二仙汤可有效阻断该作用。加味龟鹿二仙汤与顺铂合用可减轻BM-MSCs的损伤。这表明加味龟鹿二仙汤可以增强BM-MSC修复和恢复顺铂诱导的损伤组织正常功能的可能性，这可能为治疗应用提供新的方向。
To examine the exact role of flavored Guilu Erxian decoction, a Traditional Chinese Medicine (TCM) in the treatment of cisplatin-induced side-effects in bone marrow mesenchymal stem cells (BM-MSCs). BM-MSCs were isolated from bone marrow collected from SD rats and identified by flow cytometry. Cells were cultivated in MEM alpha medium containing 5% (TCM-L), 10% (TCM-M) and 20% (TCM-H) dosages of flavored Guilu Erxian decoction with or without cisplatin. Cell viability was determined through CCK-8 and thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) staining assay. Flow cytometry was used to determine cell cycle and apoptosis. The expression of p21 and cleaved-caspase-3 were examined using Western blot assay. The PI3K-AKT-mTOR pathway associated proteins, including p-PI3K, p-AKT and p-mTOR, were also examined by Western blot assay. CCK-8 and EdU staining assay demonstrated that cisplatin could inhibit cell proliferation in BM-MSCs in a dose and time dependent manner. Further, cisplatin could induce apoptosis through increasing G0/GI cell cycle arrest, p21 and cleaved-caspase-3 expression. However, these phenomena would be significantly alleviated when adding the serum containing flavored Guilu Erxian decoction. Furthermore, the PI3K-AKT-mTOR pathway activation could be inhibited by cisplatin in BM-MSCs, while flavored Guilu Erxian decoction treatment successfully abrogated this effect. Combination of flavored Guilu Erxian decoction and cisplatin could reduce the damage to BM-MSCs. This indicates that the flavored Guilu Erxian decoction can enhance the possibility of BM-MSCs repairing and rehabilitating the normal function of injured tissues induced by cisplatin, which could provide a new direction for therapeutic applications.