九香虫肽缓解槟榔碱诱导的胃粘膜上皮细胞损伤

The Role of Polypeptides from Aspongopus chinensis Dallas in Ameliorating Arecoline-induced GES-1 Cell Injury

摘要:

九香虫（Aspongopus chinensis Dallas）属于传统药食两用资源昆虫，具有多种生物学功效。该研究旨在探讨九香虫肽对槟榔碱诱导的人胃粘膜上皮细胞（GES-1）损伤的保护作用。采用九香虫肽干预槟榔碱诱导的GES-1细胞，测定抗氧化和线粒体功能等指标。结果显示，250、1 000 μg/mL的九香虫肽处理后，细胞活力分别提高了4.94%和10.89%，SOD活力分别增加了22.33%、38.29%，GSH含量增加了23.65%、40.10%；LDH活力分别下降1.51%、8.99%，ROS水平降低25.62%、48.75%。此外，九香虫肽通过回调线粒体膜电位和ATP酶活力，减轻线粒体损伤。采用转录组学技术进一步分析，发现九香虫肽主要通过调节程序性坏死通路来缓解GES-1损伤。最后利用RT-qPCR和Western-blot验证得到九香虫肽显著下调该通路p62、Hsp90、cPLA2和RIPK1、RIPK3、MLKL的表达水平（P<0.01）。综上，九香虫肽通过下调程序性坏死通路关键基因和蛋白表达，进而改善细胞氧化损伤，维持线粒体功能，缓解细胞坏死。该研究为缓解槟榔碱引发的胃黏膜损伤提供新的思路，并为九香虫肽产品开发奠定理论依据。

Abstract:

Aspongopus chinensis Dallas (ACD) is classified as a traditional medicinal and edible insect, and exhibits diverse biological effects. Polypeptides from ACD (hereinafter referred to as ACD polypeptides) were used to treat arecoline-induced GES-1 cells to clarify their protective effects against arecoline-induced GES-1 cell injury. After treatment, antioxidant and mitochondrial function indicators were evaluated. Treatment with 250 and 1 000 μg/mL of ACD polypeptides increased respective cell viability scores by 4.94% and 10.89%, superoxide dismutase activities by 22.33% and 38.29%, and glutathione content by 23.65% and 40.10%, while reducing respective lactate dehydrogenase activities by 1.51% and 8.99% and reactive oxygen species (ROS) levels by 25.62% and 48.75%. Additionally, ACD polypeptides alleviated mitochondrial damage by modulating mitochondrial membrane potential and ATPase activity. Transcriptomic analyses revealed that ACD polypeptides mitigate GES-1 cell injury by regulating necroptosis. Finally, RT-qPCR and western blot were employed to confirm that ACD polypeptides downregulate p62, Hsp90, cPLA2, RIPK1, RIPK3, and MLKL expression in this pathway (P<0.01). In summary, ACD polypeptides mitigate cellular oxidative damage, preserve mitochondrial function, and alleviate cell necrosis by downregulating key genes and protein expression in the necroptosis pathway. This study provides new insights into mitigating arecoline-induced damage to the gastric mucosa and establishes a theoretical foundation for developing ACD polypeptide products.