日本医蛭摄食前后不同组织中水蛭素基因(hirudin)的时空表达模式  

Spatiotemporal pattern of hirudin gene in Hirudo nipponia before and after feeding

作　　者：石萍[1,2,3] 游华建 邓小书 陈仕江 鲁增辉[1,2,3] SHI Pig;YOU Hua jan;DENG Xao-shu;CHEN Sijang;LU Zeng-hui(Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China;Chongqing College of Traditional Chinese Medicine,Chongqing 402760,China;Chongqing Sub-center of National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Science,Chongqing 400065,China)

机构地区：[1]重庆市中药研究院,重庆400065 [2]重庆中医药学院,重庆402760 [3]中国中医科学院中药资源中心重庆分中心,重庆400065

出　　处：《西南农业学报》2023年第7期1400-1405,共6页Southwest China Journal of Agricultural Sciences

基　　金：重庆市基本科研业务费项目(cstc2019jxjl-jbky10009);重庆市中药研究院基本科研业务费项目(jbky20210031)。

摘　　要：【目的】探究日本医蛭在不同摄食阶段、各个组织(唾液腺、肠、嗉囊、皮肤及精/卵巢)中水蛭素基因(hirudin)的时空表达模式,为水蛭药材的科学合理利用提供理论依据。【方法】基于水蛭唾液腺转录组数据筛选结果及基因克隆获得的水蛭素基因序列信息,利用实时荧光定量PCR方法,对水蛭关键活性成分的编码基因hirudin在不同摄食阶段、不同组织中的表达量进行检测分析,研究该基因的时空表达模式。【结果】水蛭素基因(hirudin)在日本医蛭的唾液腺、肠、嗉囊、皮肤及精/卵巢组织中均有表达,且在唾液腺中的表达量显著高于其他组织;Hirudin基因在不同摄食阶段的唾液腺组织中表达结果显示,摄食后第2天表达量显著升高,之后随着摄食时间的推延而呈下降趋势;Hirudin基因在嗉囊、肠中的表达量呈先升高后下降趋势,分别于摄食后第1、第2天达到最高,之后会随着停食时间延长呈下降趋势,尤其是在摄食后第7、第8天的表达量呈显著性降低;Hirudin基因在皮肤组织中也有表达,并随着停食时间的延长呈先升后降趋势,最高峰出现在摄食后第2天;在精/卵巢组织中,摄食期间hirudin基因的表达量最高,与其他摄食阶段的表达呈显著性差异。【结论】水蛭素基因(hirudin)在日本医蛭的唾液腺、肠、嗉囊、皮肤和精/卵巢组织中均有不同程度的表达,在唾液腺中的表达量显著高于其他组织。摄食行为及食物刺激可能影响水蛭素基因(hirudin)的表达,在不同摄食阶段的表达量存在显著差异。研究结果可为水蛭药材的科学采收加工、提取部位的科学选取、合理入药提供理论依据和技术支撑,对未来水蛭药材资源的可持续利用具有重要的参考价值。【Objective】In order to provide theoretical basis for the scientifc and rational utilization of leeches,the expression patterns of the hirudin gene in different tissues(salivary glands,intestines,crop,skin and genital glands)before and after feeding were analyzed.【Method】Based on the transcriptome data of the salivary of Hirudo nipponia and the cloning result of hirudin gene,the real-time fluorescent quantitative PCR method was used to detect and analyze the expression of hirudin,the coding gene for the key active ingredient of leech,in different feeding stages and different tissues,and studied the spatio-temporal expression pattern of the gene.【Result】Hirudin gene was expressed in all tissues of H.nipponia,and its expression level in salivary glands was significantly higher than those in other tissues.The expression of hirudin gene in salivary glands at different feeding stages showed a significant increase in expression on the second day after feeding,followed by a downward trend with the extension of feeding time.The expression of hirudin in crop and intestines increased at first and then decreased,and peaked on the first and second day after feeding,and then showed a downward trend with the extension of feeding time,especially on the 7~(th)and 8~(th)day after feeding.The expression of hirudin in skin increased at first and then decreased,and peaked on the second day after feeding.In genital glands tissue,the expression of hirudin was the highest during feeding,which showed significant difference from that during other feeding stages.【Conclusion】Hirudin gene is expressed in salivary glands,intestines,crops,skin and genital glands tissues to varying degrees,and its expression in salivary glands is significantly higher than that in other tissues.Both feeding behavior and food stimulation may affect the expression of hirudin gene,and there are significant differences in the expression of hirudin gene at different feeding stages.The results of the study can provide theoretical basis and technical su

关 键 词：日本医蛭 水蛭素基因(hirudin) 摄食 时空表达模式 

分 类 号：Q786[生物学—分子生物学]