酸性肽对大鼠星形胶质细胞分泌神经生长因子和脑源性神经因子水平的影响(英文)  

Influence of acidic peptide on the levels of nerve growth factor and brain-derived neurotrophic factor secreted by rat astrocytes

作　　者：安玉会[1] 赵昕[1] 马红霞[1] 毛红利[1] 谢晓燕[1] 寇现娟[1] 

机构地区：[1]郑州大学医学院生物化学与分子生物学教研室,河南省郑州市450052

出　　处：《中国临床康复》2006年第34期174-176,F0003,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：背景:神经生长因子和脑源性神经因子对神经细胞的存活和增殖非常重要。阿尔茨海默病患者的神经生长因子和脑源性神经因子水平均较低。目的:探讨酸性肽能否增加大鼠星形胶质细胞分泌神经生长因子和脑源性神经因子。设计:随机对照动物实验。单位:郑州大学医学院生物化学与分子生物学教研室。材料:实验于2003-09/2005-05在郑州大学生物活性肽研究所第一实验室和郑州大学基础医学院细胞培养中心完成。选取出生后2d内的SD乳鼠15只作为实验对象。方法:①将SD乳鼠在无菌条件下断头取出大脑皮质部分,进行星形胶质细胞的纯化培养,采用胶质纤维酸性蛋白免疫组化方法鉴定星形胶质细胞。②将培养的星形胶质细胞随机分为6组:空白对照组、血清对照组、阳性对照组、酸性肽37.5,75,150mg/L治疗组。空白对照组不施加任何实验因素,血清对照组加入体积分数为0.2的血清,阳性对照组加入1000U/mL干扰素,酸性肽治疗组则分别加入37.5,75,150mg/L的酸性肽。③将长满瓶底的第2代星形胶质细胞消化成单细胞悬液,以5×105mL等量接种于3块12孔培养板中。各组均于培养24,48,72h各时间点测定细胞存活率、细胞上清液中和细胞内神经生长因子及脑源性神经因子含量。主要观察指标:①不同培养时间各组细胞计数和存活率的检测结果。②酸性肽对大鼠星形胶质细胞增殖的影响。③不同培养时间各组星形胶质细胞上清液中神经生长因子及脑源性神经因子含量的变化。结果:①与空白对照组比较,酸性肽75,150mg/L治疗组细胞计数和细胞存活率在培养24,48,72h时均明显增加(P<0.05,0.01,0.001);酸性肽37.5mg/L治疗组虽有所增加但不明显。②与空白对照组比较,酸性肽37.5,75,150mg/L治疗组细胞增殖率均显著升高(0,17.5%,45.5%,72.5%,P<0.001)。③与空白对照组比较,酸性肽37.5,75,150mg/L治疗组细胞上清液�BACKGROUND： Nerve growth factor （NGF） and brain-derived neurotrophic factor（BDNF） are very important to the survival and proliferation of nerve cells. In the patients with Alzheimer disease （AD）, the levels of NGF and BDNF are low. OBJECTIVE： To investigate whether acidic peptide can stimulate rat astrocytes to secrete NGF and BDNF. DESIGN： A randomized control animal experiment. MATERIALS： The experiment was finished in the First Laboratory of Institute of Biopeptide, Zhengzhou University; Cellular Culture Center, School of Basic Medical Sciences of Zhengzhou University from September 2003 to May 2005. Fifteen neonatal SD rats within 2 days after birth were selected. METHODS： ①The cerebral cortex of the neonatal SD rats was removed under sterile condition, the astrocytes were isolated and cultured, and then identified with the glial fibriliary acidic protein immunohistochemical staining. ② The cultured astrocytes were randomly divided into six group： blank control group, serum control group, positive control group and acidic peptide treated groups. No treatment was given in the blank control group, serum of 0.2 in volume fraction and 1 000 U/mL interferon were added in the serum control group and positive control group, 37.5, 75 and 150 mg/L acidic peptides were added in the acidic peptide treated groups respectively. ③ The astrocytes of the 2^nd generation, which covered the whole bottom of bottle, were digested to single cell suspension, and then inoculated to three 12-well plates equally at 5×10^5/mL. The survival rate and the contents of NGF and BDNF in the superuatant of each group were determined at 24, 48 and 72 hours respectively. MAIN OUTCOME MEASURES： ① Cell numbers and survival rates at different culture time-points; ②Effect of acidic peptide on the proliferation of astrocytes in rats; ③ Changes of NGF and BDNF in the superuatant of astrocytes at different culture time-points. RESULTS： ① As compared with the blank control group, the cell numbers and survival

关 键 词：神经生长因子 脑源性神经营养因子 肽类 星形细胞 

分 类 号：R749.3[医药卫生—神经病学与精神病学]