CAJ | 학술논문

背景:目前,人脐血干细胞在脊髓损伤治疗中的研究较多,但人脐血干细胞的体外分化由许多因素所限制.蒙药广枣提取物对神经细胞有保护作用,但其作用机制不清楚.目的:观察蒙药广枣3种粗提取物促进人脐血干细胞体外分化的影响.方法:获取临床健康产妇的新鲜脐带血,制备人脐血干细胞悬液,将纯化的人脐血干细胞接种于40个培养皿中进行传代,根据蒙药广枣3种粗提取物分为:样品1组:乙醇提取,醋酸乙酯萃取,生药质量浓度为8.25 g/mL;样品2组:乙醇提取,NKA树脂分离,体积分数为10%乙醇淋洗浓缩,生药质量浓度为1.72 g/mL;样品3组:乙醇提取,NKA树脂分离,体积分数为70%乙醇淋洗浓缩,生药质量浓度为2.41 g/mL;对照组:用80%的DMEM,含体积分数为20%的小牛血清培养.观察不同质量浓度蒙药广枣提取物对人脐血干细胞增殖的影响;24,72 h后流式细胞仪测定S期的比例.结果与结论:样品3组人脐血干细胞的增殖作用不明显,10 d时样品1组和样品2组明显高于样品3组和对照组(P < 0.01).样品1组和样品2组的人脐血干细胞在24,72 h后处于S期的明显增多.提示生药质量浓度8.25,1.72 g/mL蒙药广枣粗提取物能促进人脐血干细胞的体外增殖.
배경:목전,인제혈간세포재척수손상치료중적연구교다,단인제혈간세포적체외분화유허다인소소한제.몽약엄조제취물대신경세포유보호작용,단기작용궤제불청초.목적:관찰몽약엄조3충조제취물촉진인제혈간세포체외분화적영향.방법:획취림상건강산부적신선제대혈,제비인제혈간세포현액,장순화적인제혈간세포접충우40개배양명중진행전대,근거몽약엄조3충조제취물분위:양품1조:을순제취,작산을지췌취,생약질량농도위8.25 g/mL;양품2조:을순제취,NKA수지분리,체적분수위10%을순림세농축,생약질량농도위1.72 g/mL;양품3조:을순제취,NKA수지분리,체적분수위70%을순림세농축,생약질량농도위2.41 g/mL;대조조:용80%적DMEM,함체적분수위20%적소우혈청배양.관찰불동질량농도몽약엄조제취물대인제혈간세포증식적영향;24,72 h후류식세포의측정S기적비례.결과여결론:양품3조인제혈간세포적증식작용불명현,10 d시양품1조화양품2조명현고우양품3조화대조조(P < 0.01).양품1조화양품2조적인제혈간세포재24,72 h후처우S기적명현증다.제시생약질량농도8.25,1.72 g/mL몽약엄조조제취물능촉진인제혈간세포적체외증식.
BACKGROUND: Human umbilical cord blood stem cells have been widely used in the study of spinal cord injury, but in vitro differentiation of human umbilical cord blood stem cells (hUCBSCs) has been limited by various factors. Mongalian medicine axillary choerospondias fruit extract has protective effects on neural cells, but the action mechanisms are unclear. OBJECTIVE: To observe promoting effects of 3 kinds of Mongalian medicine axillary choerospondias fruit extracts on in vitro differentiation of hUCBSCs. METHODS: Fresh umbilical cord blood was obtained from healthy puerperants to prepare hUCBSC suspension. The purified hUCBSCs were incubated in 40 petri dishes. The Mongalian medicine axillary choerospondias fruit extracts were divided into: sample 1 group: ethanol extraction, ethyl acetate extraction, crude drug mass concentration was 8.25 g/mL; sample 2 group: ethanol extraction, NKA resin isolation, 10% ethanol eluting concentration, crude drug mass concentration was 1.72 g/mL; sample 3 group: ethanol extraction, NKA resin isolation, 70% ethanol eluting concentration, crude drug mass concentration was 2.41 g/mL; control group: incubation of 80% DMEM containing 20% calf serum. Effects of various mass concentrations of Mongalian medicine axillary choerospondias fruit extract on hUCBSCs proliferation were observed. Proportion in S phase was measured using flow cytometry at 24 and 72 hours. RESULTS AND CONCLUSION: The proliferation of hUCBSCs was not significant in the sample 3 group. At day 10, the proliferation was significantly greater in the sample 1 and 2 groups compared with the sample 3 and control groups (P < 0.01). The number of hUCBSCs was significantly increased at 24 and 72 hours in S phase in the sample 1 and 2 groups. Mongalian medicine axillary choerospondias fruit extract (crude drug mass concentration 8.25, 1.72 g/mL) could promote in vitro proliferation of hUCBSCs.