What question did this study set out to answer?

The aim is to explore the role of small peptides from Periplaneta americana in mitigating H2O2-induced apoptosis in KGN cells through miRNA regulatory mechanisms.

March 3, 2026Open Access

Analysis of the resistance of small peptides from Periplaneta americana to H2O2-induced apoptosis in KGN Cells based on miRNA-seq

LXLinjie XuDali University RJRong JiangDali University JSJingjing SuFujian Medical University

Key Points

The aim is to explore the role of small peptides from Periplaneta americana in mitigating H2O2-induced apoptosis in KGN cells through miRNA regulatory mechanisms.
Used miRNA sequencing to identify differentially expressed miRNAs related to apoptosis regulation.
Conducted experiments grouping cells into control, H2O2, and H2O2 + SPPA conditions.
Screened for overlapping differentially expressed miRNAs and target genes enriched in KEGG pathways.
Verified findings using fluorescence quantitative PCR.
Identified 75 differentially expressed miRNAs between control and H2O2 groups; 30 between H2O2 and H2O2 + SPPA groups.
Found 8 upregulated and 5 downregulated overlapping miRNAs influenced by SPPA.
Identified key miRNAs playing significant roles in apoptosis resistance, including hsa-miR-103a-3p.
Established a control network detailing SPPA's anti-apoptotic mechanisms.

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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