【目的】 miRNA在昆虫的生长、发育和代谢等诸多生物学进程中发挥关键调控作用。本研究对ace-miR-3759-y进行表达和序列验证，并通过过表达与敲降ace-miR-3759-y探究其对中华蜜蜂Apis cerana cerana幼虫靶基因表达及重量的影响，以期为ace-miR-3759-y调控幼虫发育的分子机理研究提供依据。【方法】 通过Stem-loop RT-PCR和Sanger测序验证ace-miR-3759-y的表达和序列。通过饲喂模拟物(Mimic)和抑制物(Inhibitor)对中华蜜蜂幼虫血淋巴中的ace-miR-3759-y进行过表达和敲降。利用RT-qPCR检测ace-miR-3759-y的过表达和敲降效果及靶基因相对表达量。利用电子天平对幼虫进行称重。【结果】 ace-miR-3759-y在中华蜜蜂幼虫肠道和血淋巴中真实存在和表达。相较于Mimic-NC组，ace-miR-3759-y在Mimic-ace-miR-3759-y组4日龄和5日龄幼虫血淋巴中显著上调（P<0.05），在6日龄幼虫血淋巴中上调表达但组间差异不显著（P>0.05）。相较于Inhibitor-NC组，Inhibitor-ace-miR-3759-y组4、5和6日龄幼虫血淋巴中均显著下调表达（P<0.05）。过表达ace-miR-3759-y后，CIC在4、5和6日龄幼虫血淋巴中的表达量均显著下调（P<0.05）；CKI在4日龄和5日龄幼虫血淋巴中的表达量显著下调（P<0.05），在6日龄幼虫血淋巴中下调表达但组间差异不显著（P>0.05）。敲降ace-miR-3759-y后，CIC在5日龄幼虫血淋巴中显著上调表达（P<0.05），在4日龄和6日龄幼虫血淋巴中上调表达但组间差异不显著（P>0.05）；CKI在4日龄幼虫血淋巴中显著上调表达（P<0.05），在5日龄和6日龄幼虫血淋巴中上调表达但组间差异不显著（P>0.05）。此外，与Mimic-NC组相比，Mimic-miR-3759-y组的6日龄幼虫的体重下降，但组间差异不显著（P>0.05）；与Inhibitor-NC组相比，Inhibitor-miR-3759-y组的6日龄幼虫体重上升，但组间差异不显著（P>0.05）。【结论】 通过饲喂Mimic和Inhibitor的方法在中华蜜蜂幼虫血淋巴中实现了ace-miR-3759-y的有效过表达及敲降，ace-miR-3759-y负调控CIC和CKI的表达，ace-miR-3759-y对幼虫体重无显著影响。

[Objectives]  To determine the role of ace-miR-3759-y in target gene expression and body weight of Apis cerana cerana larvae in order to provide a foundation for further study of the ace-miR-3759-y-regulated molecular mechanism underlying larval development. [Methods]  The expression and sequence of ace-miR- 3759-y were confirmed by Stem-loop RT-PCR and Sanger sequencing. A. cerana cerana larvae were fed Mimic and Inhibitor to overexpress and knockdown ace-miR-3759-y in larval hemolymph. RT-qPCR was then used to determine the effect of ace-miR-3759-y overexpression and knockdown on the target genes CIC and CKI, and on larval weights which were measured using electronic scales. [Results]  ace-miR-3759-y was both present and expressed in the hemolymph of A. cerana cerana larvae. Treatment with Mimic significantly up-regulated its expression in 4- and 5-day-old larvae relative to the control group. It was also upregulated in 6-day-old larvae, but this was not significant. Conversely, treatment with Inhibitor significantly down-regulated ace-miR-3759-y expression in 4-, 5- and 6-day-old larvae relative to the control. After overexpression of ace-miR-3759-y, the expression of CIC in 4-, 5- and 6-day-old larvae was significantly down-regulated, whereas that of CKI was significantly down-regulated in 4- and 5-day-old larvae. It was also down-regulated in 6-day-old larvae, but not significantly so. Knockdown of ace-miR-3759-y significantly up-regulated the expression of CIC in 5-day-old larvae. Expression was also up-regulated in 4- and 6-day-old larvae but this was not significant. Knockdown of ace-miR-3759-y also up-regulated CKI in 4-, 5- and 6-day-old day-old larvae, although the degree of upregulation in the latter two age classes was not significant. Moreover, treatment with Mimic caused a non-significant decrease in the body weight of 6-day-old larvae relative to the Mimic control group and treatment with Inhibitor caused a non-significant increase in the body weight of 6-day-old larvae relative to the Inhibitor control group. [Conclusion]  Effective overexpression and knockdown of ace-miR-3759-y in A. cerana cerana larvae can be achieved by feeding mimic and inhibitor. ace-miR-3759-y negatively regulates expression of CIC and CKI but has insignificant effects on body weight.