Eicosanoids represent a category of oxygenated C20 polyunsaturated fatty acids that play a crucial role in regulating various physiological functions in insects. These compounds are synthesized from phospholipids through the enzymatic activity of phospholipaseA2 (PLA2). In this study, we investigated four PLA2 genes encoded in the western flower thrips, Frankliniella occidentalis, with a focus on understanding their specific functions through a variety of experimental approaches. Among the four PLA2s identified, Fo-PLA2A and Fo-PLA2B fall under the category of Ca2+-independent cellular PLA2 (iPLA2), while Fo-PLA2C and Fo-PLA2D are classified a secretory PLA2 (sPLA2). Enzyme activity assays showed that all developmental stages of the thrips exhibited significant activity against both types of PLA2 substrates (arachidonic acid (AA) and non-AA), albeit with variations in enzyme kinetics across different developmental stages. Further examination revealed that all four PLA2 genes were expressed in every developmental stage. Fo-PLA2B and Fo-PLA2C displayed higher expression levels in larvae, while Fo-PLA2A and Fo-PLA2D were predominantly expressed in female and male adults, respectively. These findings support the hypothesis that PLA2s likely serve specific functions during distinct developmental stages. Our results indicated that PLA2s play pivotal roles in mediating various physiological processes such as immunity, development, and reproduction. So, the differential expression of certain PLA2s in various stages suggests that each PLA2 may have a specific role during different stages. Additionally we conducted flouorescens in situ hybridization (FISH) analysis to localize PLA2 transcripts in different tissues, shedding light on their specific functions within these tissues. In summary, the four identified PLA2s are associated with distinct catalytic activities and exhibit differential expressions across developmental stages and tissues, collectively contributing to various physiological processes.

Phospholipase A2 (PLA2) hydrolyzes fatty acids from phospholipids at sn-2 position. Two immune-associated PLA2s are known in Spodoptera exigua. Both are calcium-independent cellular PLA2 (iPLA2A and iPLA2B). These iPLA2s are inducible to immune challenge using different epitopes such as virus, Gram+, Gram- bacteria, and fungi. However, their expression profiles are different between these two iPLA2s. Especially, immune challenge with Gram+ bacteria and fungi significantly up-regulated these iPLA2 expression, suggesting their expression under Toll signal pathway. RNA interferences of Toll or IMD signal components are being investigated.

Phospholipase A2 (PLA2) hydrolyzes phospholipids at sn-2 position to release free fatty acids. PLA2 consists of a superfamily mainly categorized as secretory PLA2 (sPLA2), cellular Ca 2+ -dependent PLA2 (cPLA2), and cellular Ca 2+ -independent PLA2 (iPLA2). We are the first to report iPLA2 in insect. Here an objective of our study is to purify a recombinant iPLA2 protein (SeiPLA2) of Spodoptera exigua using bacterial expression system. An open reading frame of SeiPLA2 was cloned into an expression vector and then transformed into Escherichia coli BL21. Over-expression with IPTG yielded recombinant SeiPLA2 (rSeiPLA2), which was then purified by an affinity chromatography using Ni-NTA column. The purified rSeiPLA2 gave significant PLA2 activity using a pyrene substrate. Its activity was inhibited by an iPLA2 specific inhibitor (BEL), but not by sPLA2 inhibitor (BPB) or cPLA2 inhibitor (MAFP

Reactive oxygen species (ROS) induces oxidation against various biomolecules including fatty acids. Calcium-independent cellular phospholipase A2 (iPLA2) has been known to function maintenance of fatty acids in phospholipids in mammals. An insect iPLA2 (Pi-iPLA2) was predicted from transciptomes of the Indianmeal moth, Plodia interpunctella. It was expressed in all developmental stages from egg to adult. RNA interference (RNAi) against Pi-iPLA2 was performed with specific double-stranded RNA (dsRNA) with more than 70% efficiency in reduction of its expression. Under this RNAi condition, P. interpuctella exhibited developmental retardation with significant accumulation of lipid peroxidation measured by the amount of melondialdehyde

Nitric oxide (NO) is an immune mediator in several insects. In addition, eicosanoids mediate various immune responses in response to microbial challenges. This study focuses on cross-talk between two immune mediators. Nitric oxide synthase (SeNOS) of Spodoptera exigua was predicted in its cDNA sequence by interrogation to the transcriptomes and validated by RT-PCR against developmental stage and larval tissues. RNA interference (RNAi) of SeNOS suppressed nodule formation of S. exigua larvae against heat-killed Xenorhabdus hominickii. RNAi of SeNOS also suppressed NO levels in fat body and hemocytes along with suppression of PLA2 activity. NOS-specific inhibitor, L-NAME, also inhibited PLA2 activity, but its enantiomer, D-NAME, did not. However, PLA2 of dexamethasone or oxindole did not suppress NO levels in the immune tissues. In addition, X hominickii did not inhibit up-regulation of NO levels. These results suggest that NO signal activates PLA2 to produce eicosanoids to mediate immune responses

Phospholipase A2 (PLA2) catalyzes the committed step for eicosanoid biosynthesis. Two cellular PLA2 genes (iPLA2 and cPLA2) have been identified in Spodoptera exigua. PLA2 activity varied among developmental stages and different tissues. There was a positive correlation between PLA2 activities and their expression levels. Interestingly, there was significant inhibition by sPLA2 inhibitor, suggesting a presence of sPLA2 in S. exigua. An entomopathogenic bacterium, Xenorhabdus hominickii, infection suppressed expression of PLA2 genes and its extract inhibited enzyme activities. Oxindole of a bacterial metabolite of X. hominikii inhibited PLA2 activity and expression.

The use of bee venom (Apis mellifera L., BV) occasionally causes side effects such as inflammation and allergic reactions in the recipients. Several case reports also suggested the treatment of BV has some limitations in its clinical uses, due to the occurrence of dermal necrosis and anaphylatic reactions. It is generally understood that bee venom allergy is mainly the result of its allergic component, phospholipase A2 (PLA2). The present study was aimed to generate PLA2-free bee venom (PBV) and evaluate its efficacy as skin care and cosmetic preparation, comparing with original bee venom (BV). Our results showed that both BV and PBV exhibited significant protective effects in UVB-irradiated human keratinocyte (HaCaT) and human dermal fibroblast (HDF) cells and they also induced type I collagen synthesis in UVB-irradiated HDF cells except BV at 3 μg/ml. Furthermore, BV and PBV showed the inhibition of UVB-stimulated matrix metalloproteinase-1 (MMP-1), a major collagen degrading enzyme in skin. However, BV, unlike PBV, exhibited strong cytotoxicities in skin cells (both HaCaT and HDF) at its working concentrations of anti-wrinkle effect. The underlying cell signaling mechanisms of anti-wrinkle effects of BV and PBV were demonstrated by the activation of ERK1/2, and p38. Conclusively, PBV appears to be the bee venom of choice with less cytotoxicity and higher efficacy on UVB-irradiated skin cells in comparison with original bee venom (BV). Therefore, PBV can better be used as a cosmetic ingredient exhibiting excellent anti-wrinkle effect against photoaging than original BV.