中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
32期
6393-6396
,共4页
刘黎军%杨雷%肖建德%王大平
劉黎軍%楊雷%肖建德%王大平
류려군%양뢰%초건덕%왕대평
许旺细胞%人参皂甙%神经生长因子
許旺細胞%人參皂甙%神經生長因子
허왕세포%인삼조대%신경생장인자
背景:人参皂甙可以促智抗衰老,保护大脑皮质运动神经元,具有抗细胞凋亡作用,但作用机制不清.目的:观察人参皂甙Rb1和Rg1对许旺细胞神经生长因子表达的影响.设计、时间及地点:细胞学体外观察,于2004-03/06在深圳市第二人民医院完成.材料:成人新鲜离体神经取自创伤离断无法再植的肢体,由深圳市第二人民医院提供.人参皂甙Rb1、Rg1由长春白求恩医科大学提供.方法:剥去神经外膜,剪成1.0～2.0 mm碎块,采用酶消化法分离许旺细胞,双30 min差速黏附法去除成纤维细胞,获得纯净度达95%以上的许旺细胞,将纯化的许旺细胞按每孔10万个细胞接种在预先涂有多聚赖氨酸的96孔细胞培养板上,设立3组:人参皂甙Rb1组分别加入含量为10,20,40,60,80 μg的人参皂甙Rb1 20 μL;人参皂甙Rg1组分别加入含量为10,20,40,60,80 μg的人参皂甙Rg1 20 μL;对照组加入PBS液20 μL.主要观察指标:流式细胞仪榆测许旺细胞神经生长因子的表达率.结果:与对照组比较,培养48 h后人参皂甙Rb1组、人参皂甙Rg1组许旺细胞神经生长因子表达率均明显升高(P＜0.05),且呈剂量依赖性增加,当人参皂甙Rb1和Rg1质量浓度达60 mg/L时许旺细胞神经生长因子表达率最高.人参皂甙Rb1及Rg1相同剂量组之间比较,许旺细胞神经生长因子表达率差异无显著件意义(P＞0.05).结论:人参皂甙Rb1和Rg1可以通过促进许旺细胞分泌神经生长因子而具有潜在加速周围神经损伤修复的作用.
揹景:人參皂甙可以促智抗衰老,保護大腦皮質運動神經元,具有抗細胞凋亡作用,但作用機製不清.目的:觀察人參皂甙Rb1和Rg1對許旺細胞神經生長因子錶達的影響.設計、時間及地點:細胞學體外觀察,于2004-03/06在深圳市第二人民醫院完成.材料:成人新鮮離體神經取自創傷離斷無法再植的肢體,由深圳市第二人民醫院提供.人參皂甙Rb1、Rg1由長春白求恩醫科大學提供.方法:剝去神經外膜,剪成1.0～2.0 mm碎塊,採用酶消化法分離許旺細胞,雙30 min差速黏附法去除成纖維細胞,穫得純淨度達95%以上的許旺細胞,將純化的許旺細胞按每孔10萬箇細胞接種在預先塗有多聚賴氨痠的96孔細胞培養闆上,設立3組:人參皂甙Rb1組分彆加入含量為10,20,40,60,80 μg的人參皂甙Rb1 20 μL;人參皂甙Rg1組分彆加入含量為10,20,40,60,80 μg的人參皂甙Rg1 20 μL;對照組加入PBS液20 μL.主要觀察指標:流式細胞儀榆測許旺細胞神經生長因子的錶達率.結果:與對照組比較,培養48 h後人參皂甙Rb1組、人參皂甙Rg1組許旺細胞神經生長因子錶達率均明顯升高(P＜0.05),且呈劑量依賴性增加,噹人參皂甙Rb1和Rg1質量濃度達60 mg/L時許旺細胞神經生長因子錶達率最高.人參皂甙Rb1及Rg1相同劑量組之間比較,許旺細胞神經生長因子錶達率差異無顯著件意義(P＞0.05).結論:人參皂甙Rb1和Rg1可以通過促進許旺細胞分泌神經生長因子而具有潛在加速週圍神經損傷脩複的作用.
배경:인삼조대가이촉지항쇠로,보호대뇌피질운동신경원,구유항세포조망작용,단작용궤제불청.목적:관찰인삼조대Rb1화Rg1대허왕세포신경생장인자표체적영향.설계、시간급지점:세포학체외관찰,우2004-03/06재심수시제이인민의원완성.재료:성인신선리체신경취자창상리단무법재식적지체,유심수시제이인민의원제공.인삼조대Rb1、Rg1유장춘백구은의과대학제공.방법:박거신경외막,전성1.0～2.0 mm쇄괴,채용매소화법분리허왕세포,쌍30 min차속점부법거제성섬유세포,획득순정도체95%이상적허왕세포,장순화적허왕세포안매공10만개세포접충재예선도유다취뢰안산적96공세포배양판상,설립3조:인삼조대Rb1조분별가입함량위10,20,40,60,80 μg적인삼조대Rb1 20 μL;인삼조대Rg1조분별가입함량위10,20,40,60,80 μg적인삼조대Rg1 20 μL;대조조가입PBS액20 μL.주요관찰지표:류식세포의유측허왕세포신경생장인자적표체솔.결과:여대조조비교,배양48 h후인삼조대Rb1조、인삼조대Rg1조허왕세포신경생장인자표체솔균명현승고(P＜0.05),차정제량의뢰성증가,당인삼조대Rb1화Rg1질량농도체60 mg/L시허왕세포신경생장인자표체솔최고.인삼조대Rb1급Rg1상동제량조지간비교,허왕세포신경생장인자표체솔차이무현저건의의(P＞0.05).결론:인삼조대Rb1화Rg1가이통과촉진허왕세포분비신경생장인자이구유잠재가속주위신경손상수복적작용.
BACKGROUND:Ginsenoside can promote wisdom,prevent aging,protect cortical motor neurons,resist cell apoptosis,but the mechanisms are unclear.OBJECTIVE:To observe the effect of Ginsenoside Rb1 and Rg1 on nerve growth factor expression in Schwann cells.DESIGN,TIME AND SETTING:The in vitro cytological study was performed at the Second People's Hospital of Shenzhen City from March to June 2004.MATERIALS:Fresh adult ex vivo nerve was obtained from limbs that were dissociated by trauma and could not be reimplanted at the Second People's Hospital of Shenzhen City.Ginsenoside Rb1 and Rg1 was supplied by the Norman Bethune University of Medical Sciences.METHODS:Epineurium was removed and cut into 1.0-2.0 mm blocks.Schwann cells were isolated by enzyme digestion.Following removing fibroblasts by double 30-minute differential attachment,Schwann cells with above 95% purity rate were harvested,and then incubated on a 96-well culture plate coated with polylysine (105 cells/well).Schwann cells in the Ginsenoside Rb1 group were subjected to 20 uL of Ginsenoside Rb1 at 10,20,40,60,80 ug.Schwann cells in the Ginsenoside Rg1 group underwent 20 uL of Ginsenoside Rg1 at 10,20,40,60,80 ug.Schwann cells in the control group were treated with 20 uL of phosphate buffered saline.MAIN OUTCOME MEASURES:Nerve growth factor expression rate was determined in Schwann cells by using flow cytometry.RESULTS:Nerve growth factor expression rate in Schwann cells was significantly increased in the Ginsenoside Rb1 and Ginsenoside Rg1 groups compared with the control group at 48 hours following incubation (P ＜ 0.05),in a dose-dependent fashion.Nerve growth factor expression rate peaked when Ginsenoside Rb1 and Ginsenoside Rg1 were 60 mg/L.No significant difference in nerve growth factor expression rate was detected between the Ginsenoside Rb1 and Ginsenoside Rg1 groups (P ＞0.05).CONCLUSION:Ginsenoside Rb1 and Rg1 has potential of promoting the recovery of damaged peripheral nerve by increasing Schwann cell producing and secreting nerve growth factor.
