银杏叶提取物对大鼠胰岛细胞获取和冻存的影响  

Extract of ginkgo biloba leaves influences the harvesting and cryopreservation of rat islet cells

作　　者：宋焕瑾[1] 薛武军[1] 丁晨光[1] 李杨[1] 田晓辉[1] 丁小明[1] 冯新顺[1] 田普训[1] 项和立[1] 靳占奎[1] 

机构地区：[1]西安交通大学医学院第一附属医院肾移植科,陕西省西安市710061

出　　处：《中国组织工程研究与临床康复》2008年第18期3437-3440,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目["GPx1-IRRS-KLK1转基因小鼠移植肾缺血损伤的保护机制研究"(30471640);"重建胰岛微环境的试验研究"(30571799)]~~

摘　　要：背景:能否获取到足够的具有生物活性的胰岛细胞是影响胰岛细胞移植成功与否的主要原因之一,细胞冻存是收集足够胰岛的可行手段,但冻存复苏过程可引起细胞的凋亡。目的:观察银杏叶提取物对大鼠胰岛获取及冻存后功能的影响。设计、时间及地点:随机对照动物实验,于2007-10/12在西安交通大学医学院环境与疾病相关基因教育部重点实验室完成。材料:选用成年SD大鼠40只用于分离提取胰岛细胞。方法:按随机数字表法将40只大鼠分为2组(n=20),银杏叶提取物组取胰腺前经尾静脉注射银杏叶提取物25mg/kg,获取胰岛后冻存,复苏后分别在含银杏叶提取物的培养基中培养3,7,14,21d(n=5)。对照组尾静脉注射等量盐水,冻存、培养时间同银杏叶提取物组。主要观察指标:电镜下观察胰岛细胞形态;应用原位末端标记法检测细胞凋亡情况;以免疫组化法检测Bcl-2表达。结果:①银杏叶提取物组胰岛细胞复苏损失率明显低于对照组(P<0.05)。②与对照组相比,银杏叶提取物组胰岛细胞复苏培养3d时内分泌颗粒及酶原颗粒增多明显。③银杏叶提取物组复苏培养21d后胰岛细胞仍可保持45%左右的存活率,对照组胰岛细胞存活率﹤35%,两组比较差异具有显著性意义(P<0.05)。④复苏培养7,14d,银杏叶提取物组胰岛细胞凋亡率低于对照组(P<0.05),Bcl-2表达高于对照组(P<0.05)。结论:银杏叶提取物能够减少胰岛冻存复苏的损失,阻止胰岛细胞的凋亡,从而保护了胰岛细胞的活性。BACKGROUND： A successful islet cell transplantation attributes to the harvesting of enough bio-active islet ceils, and cryopreservation is a feasible way to collect islet ceils, but cell apoptosis may occur in the procedure of cryopreservation and resuscitation. OBJECTIVE： To explore the influence of the extract of ginkgo biloba leaves （EGB） on rat islet ceils harvesting and function after cryopreservation. DESIGN, TIME AND SETTING： A randomized control animal study was performed in the Key Laboratory of Environment and Genes Related to Diseases （Xi＇an Jiaotong University Medical College）, Ministry of Education from October to December in 2007. MATERIALS： Forty male SD rats were used in this study to isolate islet cells. METHODS： Forty rats were divided into 2 groups at random （n=20）. In EGB group, rats were injected with 25 mg/kg EGB via tail vein, and the islet harvesting was performed. Next, the islet cells were cultured in EGB-containing medium for 3, 7, 14 and 21 days （n=5） after cryopreservation and resuscitation. In control group, an intravenous injection of equal volume of saline was given, followed by the same time of cryopreservation and resuscitation in EGB group. MAIN OUTCOME MEASURES： Cell morphology under electron microscopy; Detection of apoptosis by terminal dUTP nick end labeling （TUNEL）; Immunohistochemical detection of Bcl-2 expression. RESULTS： （1）The loss rate of ceils after resuscitation in the EGB group was obviously lower than that in the control group （P 〈 0.05）.（2）Compared with the control group, the endocrine particles and zymogen granules increased more significantly in the EGB group after islet ceils were cultured for 3 days.（3）After 21 days of resuscitation culture, 45% of islet ceils in EGB group survived, while the survival rate of ceils in the control group was less than 35%, there were significant differences between the two groups （P 〈 0.05）.（4）After 7 and 14 days of resuscitation culture, the rate of apoptosis in t

关 键 词：胰岛/细胞学 银杏素 植物提取物 低温保藏 细胞移植 

分 类 号：R617[医药卫生—外科学]