Ke Bin, Shi Lin, Xu Zhaozhong, Wu Gang, Gong Yuxin, Zhu Lixia, Wang Yanjing, Ouyang Huailiang, Wang Xilong
Department of Traditional Chinese Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Department of Traditional Chinese Medicine, Zhujiang Hospital of Southern Medical University, Guangzhou, China.
Cell Mol Biol (Noisy-le-grand). 2018 May 15;64(6):58-64.
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/G1 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.
为研究中药加味龟鹿二仙汤在治疗顺铂诱导的骨髓间充质干细胞(BM-MSCs)副作用中的确切作用。从SD大鼠采集的骨髓中分离出BM-MSCs,并通过流式细胞术进行鉴定。细胞在含有5%(中药低剂量组,TCM-L)、10%(中药中剂量组,TCM-M)和20%(中药高剂量组,TCM-H)加味龟鹿二仙汤的MEMα培养基中培养,同时设置加或不加顺铂的条件。通过CCK-8和胸腺嘧啶类似物5-乙炔基-2'-脱氧尿苷(EdU)染色试验测定细胞活力。采用流式细胞术测定细胞周期和凋亡情况。使用蛋白质免疫印迹法检测p21和裂解型半胱天冬酶-3的表达。还通过蛋白质免疫印迹法检测PI3K-AKT-mTOR信号通路相关蛋白,包括p-PI3K、p-AKT和p-mTOR。CCK-8和EdU染色试验表明,顺铂可呈剂量和时间依赖性地抑制BM-MSCs的细胞增殖。此外,顺铂可通过增加G0/G1期细胞周期阻滞、p21和裂解型半胱天冬酶-3的表达诱导细胞凋亡。然而,添加含加味龟鹿二仙汤的血清后,这些现象将得到显著缓解。此外,顺铂可抑制BM-MSCs中PI3K-AKT-mTOR信号通路的激活,而加味龟鹿二仙汤治疗成功消除了这种作用。加味龟鹿二仙汤与顺铂联合使用可减少对BM-MSCs的损伤。这表明加味龟鹿二仙汤可提高BM-MSCs修复和恢复顺铂诱导的受损组织正常功能的可能性,这可为治疗应用提供新的方向。