Fu Rong, Kong Caihua, Wang Qin, Liu Kena, Si Huaxin, Sun Ruixian, Tang Yunping, Sui Shiyan
College of Public Health, Dali University, Dali, 671000, China.
College of Agronomy and Biological Sciences, Dali University, Dali, 671003, China.
Reprod Sci. 2023 Feb;30(2):473-486. doi: 10.1007/s43032-022-01072-0. Epub 2022 Sep 9.
This study examined the protective effects of small peptides from Periplaneta americana against HO-induced mitochondrial injury in human ovarian granulosa cells. The ATP level and mitochondrial membrane potential as well as the quantity and ultrastructure of mitochondria in cells were detected. Mitochondrial DNA copy number and expression levels of Bcl2L13, LC3B, and p62 were tested. Targeted silencing of Bcl2L13 expression in KGN cells. The expression levels of Bcl2L13 and LC3B as well as interaction were evaluated. The ATP level, mtDNA-CN, and MMP of the HO group were significantly lower than those of the normal control group (P < 0.05), accompanied by a reduction in mitochondrial mass and mitochondrial fluorescence intensity (P < 0.05). However, the ATP level, mtDNA, and MMP in KGN cells were increased after SPPA treatment (P < 0.05). Scanning electron microscopy shows that SPPA ameliorates HO-induced structural damage to mitochondria. Moreover, the expression levels of Bcl2L13 and p62 in the HO group were downregulated significantly compared with those of the normal control group (P < 0.05), while LC3B was upregulated (P < 0.05). After SPPA treatment, the expression levels of Bcl2L13 and p62 were upregulated (P < 0.05), while LC3B was downregulated (P < 0.05). The Co-IP results indicated that Bcl2L13 and LC3B interacted, and this interaction was weakened after cell treatment with HO, and dissociation between Bcl2L13 and LC3B declined after SPPA treatment. SPPA inhibits KGN cell apoptosis induced by oxidative stress via inhibition of mitochondrial injury Bcl2L13-mediated mitochondrial autophagy might participate in the regulation process.
本研究检测了美洲大蠊小肽对过氧化氢（HO）诱导的人卵巢颗粒细胞线粒体损伤的保护作用。检测了细胞中的ATP水平、线粒体膜电位以及线粒体的数量和超微结构。检测了线粒体DNA拷贝数以及Bcl2L13、LC3B和p62的表达水平。对KGN细胞中Bcl2L13表达进行靶向沉默。评估了Bcl2L13和LC3B的表达水平以及相互作用。HO组的ATP水平、线粒体DNA拷贝数（mtDNA-CN）和线粒体膜电位（MMP）显著低于正常对照组（P＜0.05），同时线粒体质量和线粒体荧光强度降低（P＜0.05）。然而，经美洲大蠊小肽（SPPA）处理后，KGN细胞中的ATP水平、线粒体DNA和MMP升高（P＜0.05）。扫描电子显微镜显示，SPPA改善了HO诱导的线粒体结构损伤。此外，与正常对照组相比，HO组中Bcl2L13和p62的表达水平显著下调（P＜0.05），而LC3B上调（P＜0.05）。经SPPA处理后，Bcl2L13和p62的表达水平上调（P＜0.05），而LC3B下调（P＜co-IP结果表明，Bcl2L13与LC3B相互作用，HO处理细胞后这种相互作用减弱，SPPA处理后Bcl2L13与LC3B之间的解离减少。SPPA通过抑制线粒体损伤抑制氧化应激诱导的KGN细胞凋亡，Bcl2L13介导的线粒体自噬可能参与调节过程。
