共存递质囊泡循环动力学研究细胞模型的建立  

Establishment of cell model used by researching cycle kinetics of vesicles containing co-existence neurotransmitters

作　　者：吕军[1,2] 何功浩[1,3] 周颖[1] 侯征[1] 陈瑛瑛[1] 薛晓燕[1] 罗晓星[1] 

机构地区：[1]第四军医大学药学系药理学教研室,陕西西安710032 [2]西安交通大学医学院第一附属医院药学部,陕西西安710061 [3]成都军区昆明总医院药学部,云南昆明650032

出　　处：《西安交通大学学报（医学版）》2011年第5期557-560,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30800310);中国博士后科学基金资助项目(No.20090451521)~~

摘　　要：目的直接证明PC12细胞能够合成和分泌组胺(histamine,HA),为将其应用于交感神经共存递质囊泡循环动力学研究提供实验依据。方法常规培养PC12细胞,用嵌套RT-PCR(nRT-PCR)和Western blot技术检测其合成HA的关键酶组氨酸脱羧酶(histidine decarboxylase,HDC)的mRNA和蛋白表达,利用免疫荧光技术观测HA、HDC以及HA囊泡在细胞中的定位。结果从基因和蛋白水平均证实了该细胞中表达HDC,并利用形态学方法在细胞中检测到HA、HDC以及HA囊泡的存在。结论 PC12细胞确实含有HA,结合分化型PC12细胞与交感神经元相似的特征,可将其作为交感神经共存递质囊泡循环动力学研究的理想细胞模型。Objective To prove directly that PC12 cells can synthesize and secrete histamine（HA） and to provide experimental evidence for researching vesicle recycle kinetics of sympathetic cotransmitter.Methods PC12 cells were cultured in the conventional way.RT-PCR and Western blot techniques were used to detect histidine decarboxylase（HDC）,the key enzyme,in HA synthesis process.Immunofluorescence technique was used to detect HA,HDC and HA vesicles in the differentiated PC12 cells.Results HDC expression could be detected in PC12 cells at both the mRNA and protein levels.Furthermore,immunofluorescece data showed that detectable levels of HA,HDC and HA vesicles were found in PC12 cells.Conclusion Since PC12 cells have HA expression and the characteristic similar to that of the sympathetic neuron,they can be used as an ideal cell model for researching vesicle recycling kinetics of sympathetic cotransmitter.

关 键 词：组胺 组氨酸脱羧酶 PC12 囊泡 共存递质 嵌套RT-PCR(nRT-PCR) 交感神经元 

分 类 号：R966[医药卫生—药理学]