家蝇β-葡萄糖苷酶在Ⅲ龄幼虫消化系统以外器官的表达  

Expression of musca domestica β-glucosidase in the organs besides digestive system of Ⅲ instar larvae

作　　者：胡蓉[1] 张姝[2] 黄悦[1] 苏敏[1] 李红[1] 国果[2] 付萍[2] 吴建伟[2] 

机构地区：[1]贵州医科大学基础医学院组织学与胚胎学教研室,贵州贵阳550004 [2]贵州医科大学基础医学院人体寄生虫学教研室,贵州贵阳550004

出　　处：《局解手术学杂志》2016年第7期473-477,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：国家科技支撑计划(2011BAC06B12);国家自然科学基金(30960343;81360254)

摘　　要：目的探讨家蝇Ⅲ龄幼虫消化系统以外的器官是否产生β-葡萄糖苷酶。方法解剖显微镜下分离马氏管、气管、脂肪体和体壁等器官组织,反转录PCR检测β-葡萄糖苷酶基因是否在这些组织中表达;用原位杂交鉴定β-葡萄糖苷酶mRNA的组织分布;用纤维素酶抗体免疫组化染色检测纤维素酶的组织定位;用实时荧光定量PCR检测β-葡萄糖苷酶在各器官组织的表达差异。结果反转录PCR显示,Ⅲ龄幼虫马氏管、气管和体壁组织有β-葡萄糖苷酶基因的扩增产物;原位杂交显示,马氏管、气管和体壁上皮细胞内有β-葡萄糖苷酶mRNA分布;免疫组化结果与原位杂交定位基本一致;实时荧光定量PCR显示,β-葡萄糖苷酶mRNA表达量,马氏管>体壁>前肠>气管,差异有统计学意义(P<0.05)。结论家蝇Ⅲ龄幼虫马氏管、体壁和气管皆能分泌β-葡萄糖苷酶。结合家蝇消化系统多器官也具有分泌β-葡萄糖苷酶的特点,以家蝇β-葡萄糖苷酶为靶点的生物防治措施将有望成为减少家蝇传播人类疾病的新方法。Objective To study whether the organs besides digestive system of musca domestica Ⅲ instar larvae have the capability of produceing musca β-glucosidase. Methods Tissues of malpighian tubules,trachea,epiploon and body wall of musca domestica Ⅲ instar larvae were dissected under anatomic microscope,and the expression of β-glucosidase gene in these dissected tissues were detected by reverse transcription PCR. And the tissue localization of β-glucosidase mRNA was further identified by in situ hybridization. Moreover,anti-cellulase was used to determinate the tissue distribution with immunohistochemical staining. The relative mRNA expression levels of musca domesticaβ-glucosidase gene in these organs were tested by real-time quantitative PCR. Results The reverse transcription PCR showed that the amplification products of β-glucosidase gene were observed in tissues of malpighian tubules,trachea and body wall. β-glucosidase mRNA was shown in the epithelium cells of malpighian tubules,trachea and body wall by in situ hybridization,and it was almost the same in the results of immunohistochemical staining. The real-time quantitative PCR showed that the relative expression quantity of β-glucosidase gene in malpighian tubules and body wall were higher than that in foregut,while it was lower in itrachea than that in foregut. And it was of statistical difference in gene expression level of β-glucosidase among these organs（ P 0. 05）. Conclusion Malpighian tubules,trachea and body wall of musca domestica Ⅲ instar larvae have the function of secreting β-glucosidase. Combining with the characteristics of secreting β-glucosidase in most organs of digestive system,it may provide a new biological method for the prevention and treatment of human diseases transmitted by musca domestica with the use of taget gene β-glucosidase.

关 键 词：Β-葡萄糖苷酶 家蝇 表达 Ⅲ龄幼虫 

分 类 号：R384.21[医药卫生—医学寄生虫学]