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目的探讨黄芪注射液对多柔比星(ADR)诱导的脐血间充质干细胞(UCB-MSCs)凋亡的影响。方法收集2006年10月南华大学附属第一医院产科健康分娩的胎儿脐血,采用密度梯度与贴壁法相结合的方法采集并分离和培养UCB-MSCs,连续5次传代纯化后,经流式细胞术检测其表面标志以鉴定纯度,加入ADR(10μg/L)诱导UCB-MSCs凋亡,同时分别加入低、中、高剂量黄芪注射液(4、40、400μg/L)与UCB-MSCs共培养24 h。实验共分6组,即空白对照组,ADR组,ADR加低、中、高剂量黄芪组(4、40、400μg/L),黄芪对照组(400μg/L)。采用TUNEL法和流式细胞仪检测细胞凋亡情况。结果TUNEL法检测凋亡率黄芪对照组[(6.75±1.49)%]与空白对照组[(5.61±0.53)%]比较,无显著性差异(P>0.05);ADR组凋亡率[(39.42±2.21)%]显著高于空白对照组(P<0.01),经不同剂量黄芪注射液干预后,中、高剂量黄芪组凋亡率[(24.07±1.52)%,(22.85±2.21)%]比较无显著性差异(P>0.05),但较ADR组明显降低(Pa<0.05);低剂量黄芪组[(30.22±1.19)%]与ADR组比较变化不明显(P>0.05)。流式细胞仪检测凋亡率黄芪对照组[(5.65±0.49)%]与空白对照组[(3.71±0.17)%]比较,无显著性差异(P>0.05);ADR组凋亡率[(40.42±2.28)%]明显高于空白对照组(P<0.01);中、高剂量黄芪组凋亡率[(23.07±1.12)%,(21.85±1.21)%]均显著低于ADR组(Pa<0.05)。结论黄芪具有明显抑制ADR诱导UCB-MSCs凋亡的作用,且无剂量依赖性。
Objective To investigate the effect of astragalus injection on the apoptosis of umbilical cord blood mesenchymal stem cells (UCB-MSCs) induced by doxorubicin (ADR). METHODS: Fetal umbilical cord blood was collected from Department of Obstetrics and Health at the First Affiliated Hospital of Nanhua University in October 2006. The UCB-MSCs were collected and isolated and cultured using the method of density gradient and adherence. After passaged for 5 consecutive passages, they were purified by flow cytometry. The surface markers were used to identify the purity. AUC (10 μg/L) was added to induce apoptosis of UCB-MSCs. Simultaneously, low, middle and high doses of Huangqi injection (4, 40, 400 μg/L) were co-cultured with UCB-MSCs. 24 h. The experiment was divided into 6 groups: blank control group, ADR group, ADR plus low, medium, and high dose Huangqi group (4, 40, 400 μg/L) and Huangqi control group (400 μg/L). Apoptosis was detected by TUNEL and flow cytometry. Results Apoptosis rate was detected by TUNEL method in jaundice control group [(6.75±1.49)%] and blank control group [(5.61±0.53)%], there was no significant difference (P>0.05); ADR group apoptosis rate [(39.42 ±2.21)%] was significantly higher than that of the blank control group (P<0.01). After intervention with different doses of Huangqi injection, the apoptotic rate of medium and high dose Huangqi group [(24.07±1.52)%, (22.85±2.21)%] There was no significant difference (P>0.05), but it was significantly lower than that in the ADR group (Pa<0.05). There was no significant difference between the low-dose Astragalus group [(30.22±1.19)%] and the ADR group (P>0.05). Flow cytometry showed that the apoptosis rate of Astragalus membranaceus control group [(5.65±0.49)%] and blank control group [(3.71±0.17)%] had no significant difference (P>0.05); ADR group apoptosis rate [( 40.42±2.28)%] was significantly higher than the blank control group (P<0.01); the apoptotic rate in the medium and high dose Astragalus group was significantly lower than that in the ADR group ((23.07±1.12)%, (21.85±1.21)%). <0.05). Conclusion Astragalus membranaceus can obviously inhibit the apoptosis of UCB-MSCs induced by ADR in a dose-dependent manner.