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乙酰胆碱酯酶基因治疗猫贲门失弛缓症模型的实验研究
引用本文:徐恩斌,张忠兵,张雷,宋森涛,林勇,宁守斌,谢渭芬. 乙酰胆碱酯酶基因治疗猫贲门失弛缓症模型的实验研究[J]. 中华消化杂志, 2004, 24(6): 349-352
作者姓名:徐恩斌  张忠兵  张雷  宋森涛  林勇  宁守斌  谢渭芬
作者单位:200003,上海,第二军医大学附属长征医院消化科
摘    要:目的 研究乙酰胆碱酯酶基因转导对贲门失弛缓症模型的治疗作用。方法 将 36只家猫均分为 4组 ,第 1组为正常对照组 ;其他 3组采用胃镜下食管下端括约肌 (LES)处环形注射氯苄烷铵(BAC)的方法建立猫贲门失弛缓症模型 ,在注射BAC 8周后胃镜下LES处分别注射生理盐水 (模型对照组 )、空载病毒AdGFP(AdGFP组 )和表达乙酰胆碱酯酶 (AChE)的腺病毒AdAChET(AdAChET 组 )。1 0d后通过RT PCR和免疫组化方法检测AChE在LES中的表达 ,并观察食管压力 (LESP)的变化。结果 RT PCR证实AdAChET 组可扩增出针对目的基因的AChET 片段 ,免疫组化显示AdAChET 组在平滑肌组织中有AChE阳性表达 ,AdGFP组和模型对照组无AChE阳性表达。AdAChET 组平滑肌组织的AChE活力较模型对照组和AdGFP组高 ,差异有显著性 (P <0 .0 1 )。食管测压显示治疗后AdAChET组LESP下降较AdGFP组和模型对照组明显 ,差异有显著性 (P <0 .0 1 )。结论 编码AChET 基因腺病毒AdAChET 能松弛LES ,降低LESP ,说明AChE减少在贲门失弛缓症发生中起重要作用

关 键 词:ET  对照组  贲门失弛缓症  治疗  乙酰胆碱酯酶  LES  阳性表达  GFP  目的基因  酶基因
修稿时间:2003-09-29

Treatment of experimental cat achalasia by acetylcholinesterase gene delivery
XU En bin,ZHANG Zhong bing,ZHANG Lei,et al.. Treatment of experimental cat achalasia by acetylcholinesterase gene delivery[J]. Chinese Journal of Digestion, 2004, 24(6): 349-352
Authors:XU En bin  ZHANG Zhong bing  ZHANG Lei  et al.
Affiliation:XU En bin,ZHANG Zhong bing,ZHANG Lei,et al. Department of Gastroenterology,Changzheng Hospital,Second Military Medical University,Shanghai 200003,China
Abstract:Objective To investigate the therapeutic effect transferring acetylcholinesterase (AChE) gene into experimental cat achalasia. Methods Thirty six cats were divided into 4 groups.Group 1 was served as healthy control.The cats in other three groups were injected with benzaikoniumchloride(BAC) into the lower esophageal sphincter(LES) under endoscopy to establish achalasia model. Then the cats were injected with normal saline (group 2 as experiment control),adenoviral vectors containing green fluorescent protein (AdGFP, group 3) or adenoviruses containing full length AChE T cDNA (group 4) by the same way 8 weeks later. RT PCR and immunohistochemistry was performed to detect the expression of the AChE 10 days later.Manometry was carried out to check the pressure of LES. Results RT PCR amplification results indicated that AChE T fragment corresponding to target gene was presented in group 4. The expression of AChE T mRNA was positive and AChE protein could be detected by immunohistochemistry in the LES of cats treated with AdAChE T. In addition AChE activity increased significantly in group 4 than that in group 3 and group 2( P <0.01). Manometrically,the LES of AdAChE T treated cats showed lower pressures than that in the other( P <0.01). Conclusions The adenovirus encoding AChE T gene delivery in vivo is able to reduce the pressure of LES. The decrease of AChE may be associated with the development of achalasia.
Keywords:Acetylcholinesterase  Achalasia  Gene therapy
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