中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2006年
34期
174-176,封三
,共4页
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2 d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37.5,75,150 mg/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1 000 U/mL干扰素,酸性肽治疗组则分别加入37:5,75,150 mg/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24,48,72 h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75,150 mg/L治疗组细胞计数和细胞存活率在培养24,48,72 h时均明显增加(P＜0.05,0.01,0.001);酸性肽37.5 mg/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37.5,75,150 mg/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37.5,75,150 mg/L治疗组细胞上清液中神经生长因子的吸光度值在培养24,48,72 h时均明显增加(P＜0.001);除了在培养24 h时间点酸性肽37.5 mg/L治疗组不能增加星形胶质细胞分泌脑源性神经因子外,其他各浓度酸性肽治疗组在培养24,48,72 h时脑源性神经因子的吸光度值均显著提高(P＜0.05,0.001).结论:酸性肽能够不同程度地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.
揹景:神經生長因子和腦源性神經因子對神經細胞的存活和增殖非常重要.阿爾茨海默病患者的神經生長因子和腦源性神經因子水平均較低.目的:探討痠性肽能否增加大鼠星形膠質細胞分泌神經生長因子和腦源性神經因子.設計:隨機對照動物實驗.單位:鄭州大學醫學院生物化學與分子生物學教研室.材料:實驗于2003-09/2005-05在鄭州大學生物活性肽研究所第一實驗室和鄭州大學基礎醫學院細胞培養中心完成.選取齣生後2 d內的SD乳鼠15隻作為實驗對象.方法:①將SD乳鼠在無菌條件下斷頭取齣大腦皮質部分,進行星形膠質細胞的純化培養,採用膠質纖維痠性蛋白免疫組化方法鑒定星形膠質細胞.②將培養的星形膠質細胞隨機分為6組:空白對照組、血清對照組、暘性對照組、痠性肽37.5,75,150 mg/L治療組.空白對照組不施加任何實驗因素,血清對照組加入體積分數為0.2的血清,暘性對照組加入1 000 U/mL榦擾素,痠性肽治療組則分彆加入37:5,75,150 mg/L的痠性肽.③將長滿瓶底的第2代星形膠質細胞消化成單細胞懸液,以5×105mL等量接種于3塊12孔培養闆中.各組均于培養24,48,72 h各時間點測定細胞存活率、細胞上清液中和細胞內神經生長因子及腦源性神經因子含量.主要觀察指標:①不同培養時間各組細胞計數和存活率的檢測結果.②痠性肽對大鼠星形膠質細胞增殖的影響.③不同培養時間各組星形膠質細胞上清液中神經生長因子及腦源性神經因子含量的變化.結果:①與空白對照組比較,痠性肽75,150 mg/L治療組細胞計數和細胞存活率在培養24,48,72 h時均明顯增加(P＜0.05,0.01,0.001);痠性肽37.5 mg/L治療組雖有所增加但不明顯.②與空白對照組比較,痠性肽37.5,75,150 mg/L治療組細胞增殖率均顯著升高(0,17.5%,45.5%,72.5%,P＜0.001).③與空白對照組比較,痠性肽37.5,75,150 mg/L治療組細胞上清液中神經生長因子的吸光度值在培養24,48,72 h時均明顯增加(P＜0.001);除瞭在培養24 h時間點痠性肽37.5 mg/L治療組不能增加星形膠質細胞分泌腦源性神經因子外,其他各濃度痠性肽治療組在培養24,48,72 h時腦源性神經因子的吸光度值均顯著提高(P＜0.05,0.001).結論:痠性肽能夠不同程度地增加大鼠星形膠質細胞分泌神經生長因子和腦源性神經因子.
배경:신경생장인자화뇌원성신경인자대신경세포적존활화증식비상중요.아이자해묵병환자적신경생장인자화뇌원성신경인자수평균교저.목적:탐토산성태능부증가대서성형효질세포분비신경생장인자화뇌원성신경인자.설계:수궤대조동물실험.단위:정주대학의학원생물화학여분자생물학교연실.재료:실험우2003-09/2005-05재정주대학생물활성태연구소제일실험실화정주대학기출의학원세포배양중심완성.선취출생후2 d내적SD유서15지작위실험대상.방법:①장SD유서재무균조건하단두취출대뇌피질부분,진행성형효질세포적순화배양,채용효질섬유산성단백면역조화방법감정성형효질세포.②장배양적성형효질세포수궤분위6조:공백대조조、혈청대조조、양성대조조、산성태37.5,75,150 mg/L치료조.공백대조조불시가임하실험인소,혈청대조조가입체적분수위0.2적혈청,양성대조조가입1 000 U/mL간우소,산성태치료조칙분별가입37:5,75,150 mg/L적산성태.③장장만병저적제2대성형효질세포소화성단세포현액,이5×105mL등량접충우3괴12공배양판중.각조균우배양24,48,72 h각시간점측정세포존활솔、세포상청액중화세포내신경생장인자급뇌원성신경인자함량.주요관찰지표:①불동배양시간각조세포계수화존활솔적검측결과.②산성태대대서성형효질세포증식적영향.③불동배양시간각조성형효질세포상청액중신경생장인자급뇌원성신경인자함량적변화.결과:①여공백대조조비교,산성태75,150 mg/L치료조세포계수화세포존활솔재배양24,48,72 h시균명현증가(P＜0.05,0.01,0.001);산성태37.5 mg/L치료조수유소증가단불명현.②여공백대조조비교,산성태37.5,75,150 mg/L치료조세포증식솔균현저승고(0,17.5%,45.5%,72.5%,P＜0.001).③여공백대조조비교,산성태37.5,75,150 mg/L치료조세포상청액중신경생장인자적흡광도치재배양24,48,72 h시균명현증가(P＜0.001);제료재배양24 h시간점산성태37.5 mg/L치료조불능증가성형효질세포분비뇌원성신경인자외,기타각농도산성태치료조재배양24,48,72 h시뇌원성신경인자적흡광도치균현저제고(P＜0.05,0.001).결론:산성태능구불동정도지증가대서성형효질세포분비신경생장인자화뇌원성신경인자.
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.MATERLALS: The experiment was finished in the First Laboratory of Institute of Biopeptide, Zhengzhou University; Cellular Culture Center,School of Basic Medical Sciences of Zhe ngzhou University from September 2003 to May 2005. Fifteen neonatal SD rats within 2 days after birth were selected.METHODS: ① The 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 2nd generation, which covered the whole bottom of bottle, were digested to single cell suspension, and then inoculated to three 12-well plates equally at 5×105 /mL. The survival rate and the contents of NGF and BDNF in the supernatant 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; ② Effect of acidic peptide on the proliferation of astrocytes in rats; ③ Changes of NGF and BDNF in the supernatant of astrocytes at different culture time-points.RESULTS: ① As compared with the blank control group, the cell numbers and survival rates at 24, 48 and 72 hours were obviously increased in the acidic peptide groups treated with 75 and 150 mg/L (P＜0.05, 0.01,0.001), but not obviously increased in the acidic peptide group treated with 37.5 mg/L. ② As compared with the blank control group, the rates of proliferation in the acidic peptide groups treated with 37.5, 75 and 150 mg/L were all significantly increased (17.5%, 45.5%, 72.5%, P＜0.001). ③ As compared with the blank control group, the absorbance (A) values of NGF in the supernatant at 24, 48 and 72 hours were all markedly increased in the acidic peptide groups treated with 37.5, 75 and 150 mg/L (P＜0.001),and the A values of BDGF in the supernatant at 48 and 72 hours were significantly increased (P＜0.05, 0.01).CONCLUSION: Acidic peptide can increase the secretions of NGF and BDNF of rat astrocytes to different extent.
