Abstract
Apoptosis of ovarian granular cells is closely related with weakening fertility of women. Hence, resisting apoptosis of human ovarian granular cells (KGN cells) is of important significance. According to studies, detection of oxidation indicators such as ROS, NO, MDA and SOD, DAPI fluorescence staining experiment, mRNA and Western Blot test of Cleaved-caspase-3, Caspase-3, Bax and Bcl-2 demonstrate that small peptides from Periplaneta americana (SPPA) can improve H2O2-induced apoptosis of KGN cells. While SPPA has been reported to possess antioxidant potential, its specific role and molecular mechanisms within ovarian granulosa cells-particularly regarding its involvement in the miRNA regulatory network have not yet been thoroughly investigated. Therefore, high-throughput sequencing of miRNAs was employed to identify differentially expressed miRNAs (DEMs) that are associated with the regulatory mechanisms through which SPPA inhibits H2O2-induced apoptosis in KGN cells. Experiments were divided into three groups, namely, the control group, H2O2 group and H2O2 + SPPA group. A total of 75 DEMs were screened by comparing the control group and the H2O2 group, and 30 DEMs were screened by comparing the H2O2 group and H2O2 + SPPA group. It is important to note that 8 overlapping DEMs were identified upregulating in H2O2 exposure, but downregulating in SPPA repair. Another 5 overlapping DEMs were identified downregulating in H2O2 exposure, but upregulating in SPPA repair. 3534 target genes of the aforementioned 13 DEMs have significant enrichment in multiple KEGG pathways. Among them, hsa-miR-103a-3p, NovelmiRNA-214 and NovelmiRNA-383 play significant roles in SPPA resistance process of cell apoptosis. 5 DEMs were selected for florescence quantitative PCR (qPCR) verification and the expression level was consistent with sequencing results. Finally, a control network of SPPA resistance against the H2O2-induced KGN cell apoptosis was built based on the target genes screened by the miRNA-seq technology. This study provides a direction and some references to screen and suggest miRNAs that may be involved in the anti-apoptotic process of SPPA.
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Data availability
The miRNA-seq raw data is deposited on the Sequence Read Archive (SRA) database at https://www.ncbi.nlm.nih.gov/bioproject, BioProject: PRJNA837735.
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Acknowledgements
We would like to thank Suzhou Jinweizhi Biotechnology Co., Ltd for miRNA-seq data analysis. Additionally, thanks to figdraw (https://www.figdraw.com/static/index.html#/) for providing the drawing platform and everyone who was involved in this study.
Funding
This work was supported by National Natural Science Foundation of China (NO. 32260865); Yunnan Province Young and Middle-aged Academic and Technical Leaders Reserve Talents Project (NO. 202305AC160037), Wang Junjun Expert Workstation Project of Yunnan Province (NO. 202305AF150083); Basic Research Program of Yunnan Provincial Department of Science and Technology (NO. 202101BA070001-202).
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Linjie Xu: conceptualization, data curation, methodology, project administration, resources, software, supervision, validation, writing-original draft. Rong Jiang: data curation, formal analysis, investigation, methodology, resources, software, validation, writing-original draft. Jingjing Su: data curation, investigation, resources, visualization, writing-review & editing. Ruixian Sun: formal analysis, methodology, resources, supervision, writing-original draft. Lilian Yang: formal analysis, project administration, resources. Yunping Tang: formal analysis, resources. Shiyan Sui: conceptualization, funding acquisition, project administration, resources, supervision, validation, writing-review & editing. All authors have read and agreed to the published version of the manuscript.
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Xu, L., Jiang, R., Su, J. et al. Analysis of the resistance of small peptides from Periplaneta americana to H2O2-induced apoptosis in KGN Cells based on miRNA-seq. Sci Rep (2026). https://doi.org/10.1038/s41598-026-41839-y
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DOI: https://doi.org/10.1038/s41598-026-41839-y
