Wrońska Anna Katarzyna, Kaczmarek Agata, Kazek Michalina, Boguś Mieczysława Irena
Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Warsaw, Poland.
BIOMIBO, Warsaw, Poland.
Front Physiol. 2022 Jan 6;12:774086. doi: 10.3389/fphys.2021.774086. eCollection 2021.
Apoptosis and autophagy, the mechanisms of programmed cell death, play critical roles in physiological and pathological processes in both vertebrates and invertebrates. Apoptosis is also known to play an important role in the immune response, particularly in the context of entomopathogenic infection. Of the factors influencing the apoptotic process during infection, two of the lesser known groups are caspases and eicosanoids. The aim of this study was to determine whether infection by the entomopathogenic soil fungus is associated with apoptosis and changes in caspase activity in the hemocytes of larvae, and to confirm whether fungal infection may affect eicosanoid levels in the host. Larvae were exposed for 24 h to fully grown and sporulating fungus. Hemolymph was collected either immediately after termination of exposure (F24 group) or 24 h later (F48 group). Apoptosis/necrosis tests were performed in hemocytes using fluorescence microscopy and flow cytometry, while ELISA tests were used to measure eicosanoid levels. Apoptosis and necrosis occurred to the same degree in F24, but necrosis predominated in F48. Fungal infection resulted in caspase activation, increased PGE1, PGE2, PGA1, PGF2α, and 8-iso-PGF2α levels and decreased TXB2 levels, but had no effect on TXA2 or 11-dehydro-TXB2 concentrations. In addition, infected larvae demonstrated significantly increased PLA2 activity, known to be involved in eicosanoid biosynthesis. Our findings indicate that fungal infection simultaneously induces apoptosis in insects and stimulates general caspase activity, and this may be correlated with changes in the concentrations of eicosanoids.
细胞凋亡和自噬作为程序性细胞死亡机制,在脊椎动物和无脊椎动物的生理及病理过程中都发挥着关键作用。细胞凋亡在免疫反应中也起着重要作用,尤其是在昆虫病原感染的情况下。在感染过程中影响细胞凋亡进程的因素中,半胱天冬酶和类花生酸是两个鲜为人知的类别。本研究的目的是确定昆虫病原土壤真菌的感染是否与幼虫血细胞中的细胞凋亡及半胱天冬酶活性变化相关,并确认真菌感染是否会影响宿主体内类花生酸水平。将幼虫暴露于完全生长并产孢的真菌中24小时。在暴露结束后立即(F24组)或24小时后(F48组)收集血淋巴。使用荧光显微镜和流式细胞术对血细胞进行细胞凋亡/坏死检测,同时使用酶联免疫吸附测定法测量类花生酸水平。F24组中细胞凋亡和坏死程度相同,但F48组中坏死占主导。真菌感染导致半胱天冬酶激活,PGE1、PGE2、PGA1、PGF2α和8-异-PGF2α水平升高,TXB2水平降低,但对TXA2或11-脱氢-TXB2浓度无影响。此外,受感染的幼虫表现出显著增加的PLA2活性,已知该酶参与类花生酸生物合成。我们的研究结果表明,真菌感染同时诱导昆虫细胞凋亡并刺激总体半胱天冬酶活性,这可能与类花生酸浓度的变化相关。