气体信号分子硫化氢调控家蚕生长发育的机理研究

Hydrogen sulfide (H2S) is the 3rd gaseous signaling molecule that was found after NO and CO.H2S displayed Multiple important functions just called ‘Do It All’. Exposure to H2S could prolong the lifespan, improve heat resistance, drought tolerance and reproductive capacity of Drosophila melanogaster and Caenorhabditis elegans. There are no reports about the regulation of H2S on the growth and development of silkworm to date. Our previous research demonstrated that the appropriate concentration of H2S treatment can effectively prolong the 5th instar larvae and pupal stage, induce the increase of the systolic pressure in the midgut, increase the amount of the whole cocoon, cocoon shell and eggs. In present study, H2S will be used to regulate the growth and development of silkworm; and then, the reasonable concentration of H2S treatment of silkworm will be determined; the corresponding biochemical, metabolic changes and the key signaling pathways of H2S regulating the development time of silkworm also will be identified. The implementation of the project will be the first time to analyze the regulations of H2S on the growth of silkworm, which is significant important to enrich the theory and application of gas signal molecules in regulating life, will provide a theoretical basis for further development H2S for sericulture growth regulator, and will play important roles to achieve high yield, high quality and stable production of sericulture.

硫化氢（H2S）是继NO 与CO 之后，在动植物体中被发现的第3种功能性气体信号分子。H2S在动植物体内，被证明具有“Do It All（全能）”的功能。H2S在果蝇和线虫中具有延长寿命、提高耐热性、耐旱性和生殖能力的功能。H2S调控家蚕生长发育至今尚没有报道。前期我们已经初步探明合适浓度的H2S处理可以有效延长5龄幼虫的龄期和蛹期、诱导中肠收缩压的增强、增加全茧量、茧层量和产卵量。本项目开展H2S调控家蚕的生长发育研究，确定H2S合理的作用浓度，明确H2S调控家蚕的生化和代谢过程，鉴定H2S调控家蚕发育时间的关键信号通路。项目的实施，率先开展H2S气体信号分子对家蚕生长的调控，对于丰富气体信号分子调控生命体的理论和应用具有一定的意义；对于把H2S开发为养蚕的生长调节剂提供理论基础，实现养蚕业的高产、高质和稳产具有重要的支撑作用。

硫化氢（H2S）作为一种新型气体信号分子，在维持体内稳态和调节生理过程中起重要作用。本项目结果表明硫化氢处理家蚕呈现典型的“双面性”，50 μM以上的浓度H2S对家蚕有显著毒害，低浓度H2S能增加家蚕的平均体重、全茧量、茧层量和茧层率，但对家蚕的发育时间和生殖力无显著影响；熏蒸法处理家蚕的H2S的最优浓度为7.5 μM。H2S对不同品系的家蚕体重和茧质促进作用顺序为：地方及多化性＞欧系＞中系＞日系。利用LC-MS的代谢组学技术分析表明7.5 μM H2S处理后，家蚕5龄3天的血淋巴共鉴定出45个差异代谢物，其中25个上调，20个下调；差异代谢物中（6Z, 9Z, 12Z）-十八碳三烯酸、苹果酸、磷酸胆碱等含量升高，棕榈酸、尿酸盐等含量下降；差异代谢物主要影响嘧啶代谢、嘌呤代谢、脂肪酸代谢、线粒体中的脂肪酸延伸等代谢途径。转录组学、miRNA组学和定量PCR结果表明H2S处理后家蚕脂肪体内差异表达基因主要富集在内吞作用、糖酵解/糖异生、TCA循环、NF-κB信号通路、VEGF信号通路、TNF信号通路等转录信号通路。进一步采用RNAi的方法，合成相关基因的siRNA，敲除内吞作用通路的Vps4基因和糖酵解/糖异生通路通路的PGM基因，结合H2S处理，结果说明H2S可以通过调控内吞作用通路、糖酵解/糖异生通路实现对家蚕生长发育的调控。本项目的结果对于丰富气体信号分子调控生命体的理论和应用具有重要的科学意义，为把H2S开发为养蚕的生长调节剂提供了理论基础，对实现养蚕业的高产、高质和稳产具有重要的支撑作用。..