摘要
目的探讨miR-276-3p在白纹伊蚊幼虫几丁质代谢中的作用,为实现利用几丁质代谢相关miRNA防控白纹伊蚊提供新的靶标。方法合成miR-276-3p特异性模拟物agomir和抑制剂antagomir的壳聚糖纳米颗粒,通过饲喂法过表达或抑制白纹伊蚊幼虫体内miR-276-3p的表达。利用反转录实时荧光定量PCR(RT-qPCR)检测幼蚊体内miR-276-3p的相对表达量,利用生化分析法测定幼蚊及蛹的几丁质含量变化,并统计幼蚊存活率、化蛹率及蛹的羽化率。表达量差异比较使用two-way方差分析,几丁质含量两组间比较使用t检验,存活率差异分析采用乘积极限法的log-rank检验和Mantel-Cox检验。结果miR-276-3p agomir壳聚糖纳米颗粒可有效过表达白纹伊蚊幼虫体内miR-276-3p的表达量,第1和第3天后表达量较对照分别上调73.96%(F=932.401,P<0.001)、16.95%(F=114.551,P=0.047);miR-276-3p antagomir壳聚糖纳米颗粒可有效抑制miR-276-3p的表达,第1和第5天后表达量较对照分别下调13.42%(F=293.632,P<0.001)和1.49%(F=69.542,P=0.014);过表达miR-276-3p导致幼虫几丁质含量上升了25.34%(t=85.838,P=0.019),幼虫存活率下降,第8天较对照组差异有统计学意义(χ^(2)=77.490,P=0.020),化蛹率为70.00%,低于对照组的90.00%;抑制miR-276-3p表达导致白纹伊蚊幼虫与蛹的几丁质含量分别下降了40.06%(t=71.811,P=0.028)和46.82%(t=92.147,P=0.011)。结论miR-276-3p对于白纹伊蚊幼虫的几丁质含量、存活率及化蛹率存在显著影响,相关研究结果为利用miRNAs调控白纹伊蚊幼虫的几丁质代谢,进而实现蚊虫防控提供了基础资料。
Objective To investigate the role of miR-276-3p in the chitin metabolism of Aedes albopictus larvae,so as to provide a new target for the control of Ae.albopictus using chitin metabolism-related miRNAs.Methods miR-276-3p-specific mimic agomir and inhibitor antagomir chitosan nanoparticles were synthesized to overexpress and suppress the expression of miR-276-3p in Ae.albopictus larvae through feeding,respectively.The relative expression of miR-276-3p in larvae was measured by reverse transcription quantitative real-time PCR.The chitin levels of larvae and pupae were determined using biochemical assay.The survival rate and pupation rate of larvae and the eclosion rate of pupae were calculated.Two-way ANOVA was used for mirR-276-3p expression differences analysis,T-test was used for chitin content comparison between different groups,and the log-rank test and Mantel-Cox test were used for survival rate difference analysis.Results miR-276-3p agomir chitosan nanoparticles effectively overexpressed miR-276-3p in Ae.albopictus larvae,the expression on the 1st and 3rd days increased by 73.96%(F=932.401,P<0.001)and 16.95%(F=114.551,P=0.047)respectively,while miR-276-3p antagomir chitosan nanoparticles significantly suppressed the expression of miR-276-3p in Ae.albopictus larvae,the expression on the 1st and 5th days decreased by 13.42%(F=293.632,P<0.001)and 1.49%(F=69.542,P=0.014)respectively.The overexpression of miR-276-3p increased the chitin content by 25.34%(t=85.838,P=0.019).The survival rate decreased significantly,and the difference between the experimental group and the control group on the 8th day was statistically significant(χ^(2)=77.490,P=0.020),and the pupation rate was 70.00%,which was 90.00%and lower than that of the control group.The suppression of miR-276-3p reduced chitin levels in the larvae by 40.06%(t=71.811,P=0.028)and pupae by 46.82%(t=92.147,P=0.011)of Ae.albopictus.Conclusions miR-276-3p has significant effects on the chitin content,survival rate,and pupation rate of Ae.albopictus larvae.This st
作者
傅菁文
刘文娟
张忠
张瑞玲
FU Jing-wen;LIU Wen-juan;ZHANG Zhong;ZHANG Rui-ling(School of Clinical and Basic Medical Sciences,Shandong First Medical University&Shandong Academy of Medical Sciences,Ji’nan,Shandong 250000,China;School of Laboratory Animal&Shandong Laboratory Animal Center,Shandong First Medical University&Shandong Academy of Medical Sciences,Ji’nan,Shandong 250000,China)
出处
《中国媒介生物学及控制杂志》
CAS
北大核心
2023年第5期589-595,共7页
Chinese Journal of Vector Biology and Control
基金
国家自然科学基金(81871686)。
