摘要
目的探讨黄连素体外诱导人脐带间充质干细胞(hUMSCs)向神经细胞分化的作用。方法体外对hUMSCs进行培养,以不同浓度黄连素(分4组:对照组即0 mg/L组、50 mg/L组、100 mg/L组和200 mg/L组)诱导其向神经样细胞分化,显微镜下观察细胞形态改变并记录,并通过细胞免疫化学及免疫荧光技术检测细胞表面神经标志物巢蛋白(Nestin)、神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)的表达。组间比较采用重复测量数据的方差分析。结果经黄连素诱导72 h,细胞呈典型神经细胞样形态,且80%以上细胞表达神经细胞特异性标志物Nestin、NSE、GFAP。不同药物浓度作用下,神经样细胞分化率随时间变化情况为,200 mg/L组阳性细胞率在诱导48 h即可达峰值,高于100 mg/L组和50 mg/L组[(81.4±5.8)%比(50.4±4.7)%比(3.6±2.3)%,F=25.806,P<0.05],差异有统计学意义,但诱导72 h后明显下降,阳性表达细胞出现漂浮凋亡;100 mg/L组阳性细胞率在诱导72 h达到峰值,高于200 mg/L组和50 mg/L组[(87.7±4.6)%比(57.7±7.7)%比(14.3±5.3)%,F=24.815,P<0.05],差异有统计学意义,且高表达可持续维持3 d,诱导120 h后仍高达(85.2±5.0)%;50 mg/L组阳性细胞率一直相对较低,在诱导6 d达到峰值,低于100 mg/L组、高于200 mg/L组[(15.8±6.5)%比(64.5±6.6)%比(8.3±4.0)%,F=22.516,P<0.05],差异有统计学意义。结论黄连素体外可诱导hUMSCs向神经样细胞分化,浓度为100μg/ml时诱导最为明显,且存活时间最长。
Objective To investigate the effects of berberine on the differentiation of human umbilical cord mesenchymal stem cells(hUMSCs)into nerve cells in vitro.Methods HUMSCs were cultured in vitro.The berberine at different concentrations was added to induce hUMSCs to differentiate(divided into four groups:control group,50 mg/L,100 mg/L and 200 mg/L berberine groups).Cell morphology was observed dynamically.The expression of the neural markers including Nestin,neuron specific enolase(NSE)and glial fibrillary acidic protein(GFAP)was detected by immunohistochemistry and immunofluorescence staining.Results At 72 h after the induction with berberine,the induced cells displayed a classical neural shape,and more than 80%of them expressed neural markers,including Nestin,NSE and GFAP.At different induction concentrations,the positive cell rate of neuron-like cells varied at different time points:the rate of positive cells in the 200 mg/L berberine group reached its peak after 48 h of induction,higher than that in the 100 mg/L and 50 mg/L berberine group[(81.4±5.8)%vs.(50.4±4.7)%vs.(3.6±2.3)%,F=25.806,P<0.05],but decreased significantly after 72 h of induction,and the positive expression cells apoptotic.The rate of positive cells in the 100 mg/L berberine group reached its peak after 72 h of induction,higher than that in the 200 mg/L and 50 mg/L berberine group[(87.7±4.6)%vs.(57.7±7.7)%vs.(14.3±5.3)%,F=24.815,P<0.05],and the high expression was sustained for 3 days.After 120 h of induction,the rate of positive cells was still very high[(85.2±5.0)%].However,the rate of positive cells in the 50 mg/L berberine group remained relatively low and reached a peak at 6th day after induction,lower than that in the 100 mg/L berberine group and higher than that in the 50μg/ml berberine group[(15.8±6.5)%vs.(64.5±6.6)%vs.(8.3±4.0)%,F=22.516,P<0.05].Conclusion Baicaline can induce HUMSCs to differentiate into neuro-like cells in vitro.The induction potential of berberine by a concentration of 100 mg/L is the most obvious and the lo
作者
闫冬冬
兰海涛
宋国强
南成睿
石云鹏
李凤菊
赵宗茂
Yan Dongdong;Lan Haitao;Song Guoqiang;Nan Chengrui;Shi Yunpeng;Li Fengju;Zhao Zongmao(Department of Neurosurgery,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Radiology,Hebei Provincial Corps Hospital of Chinese People’s Armed Police Forces,Shijiazhuang 050050,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第1期71-75,共5页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81870984)
国家自然科学基金青年基金项目(81401032)
河北省自然科学基金(H2015206309)
河北省国际科技合作基地建设专项(193977143D)
河北省科技计划项目(132077125D)。
关键词
人脐带间充质干细胞
黄连素
分化
神经样细胞
Human umbilical cord mesenchymal stem cells
Berberine
Differentiation
Neuron-like cells
