人S-100β体外表达及其对神经元细胞增殖作用的研究  

Human S-100β expressed in vitro and its role on proliferation of cultured SK-N-SH neuron

作　　者：汪凯[1] 汪渊[2] 马传庚[3] 周青[2] 刘虎[2] 尹世杰[1] 苏启庚[1] 

机构地区：[1]安徽医科大学第一附属医院神经内科,合肥230022 [2]安徽医科大学第一附属医院分子生物学实验室,合肥230022 [3]安徽医科大学第一附属医院药理学教研室,合肥230022

出　　处：《中国药理学通报》2003年第7期784-787,共4页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金资助课题 (No 0 10 4 36 0 2 );优秀青年科技基金资助课题 (No 2 0 0 2 [0 2 ] )

摘　　要：目的　构建人S 1 0 0 β表达载体 ,体外表达人重组S 1 0 0 β ,观察S 1 0 0 β的神经营养作用。方法　应用RT PCR合成人S 1 0 0 βcDNA ,插入质粒 pBluescriptⅡSK(+)中 ,转化入大肠杆菌XL1 Blue中并在体外获得表达。以镜下形态学及酸性磷酸酶测定法观察表达蛋白对体外培养的SK N SH神经元增殖的作用。结果　重组质粒体外表达的人S 1 0 0 β可促进SK N SH神经元细胞附壁增加 ,体积增大 ,数目增多 ,突起增多延长 ;且明显增加培养细胞酸性磷酸酶活性。结论　S 1 0 0 β对体外培养的SK N SH神经元增殖有促进作用。AIM To construct human S 100β recombinant vector, purify and identify the target expressed S 100β protein and conduct preliminary study into the neurotrophic activity of S 100β. METHODS mRNA was obtained from human brain first. Then we got human S 100β cDNA by RT PCR method and inserted cDNA into pBluescript Ⅱ SK(+) vector. The combinative vector was then transformed into ecoli (XL1 blue). The expressed protein was purified by DEAE 52 and phenyl sepharose CL 4B affinity chromatography, and identified with SDS PAGE and Western Blot method. The neurotrophic activity of S 100β in cultured SK N SH neuron was examined with the indicator of morphology and acid phosphatase assay (APA). RESULTS The expressed protein of recombinant vector is verified as S 100β. The results showed that S 100β increased the number of cell and volume of cultured SK N SH neuron and increased the activity of acid phosphatase of neuron obviously. CONCLUSION S 100β promotes the proliferation of SK N SH neuron in vitro .

关 键 词：S-100 聚合酶链式反应 WESTERN-BLOT 细胞培养 神经元 增殖 

分 类 号：R-332[医药卫生] R329.24