Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
Bioengineered. 2022 Jan;13(1):217-226. doi: 10.1080/21655979.2021.2012908.
Octreotide is a synthetic octapeptide of natural somatostatin. We aimed to investigate the influence of Octreotide on lipopolysaccharide (LPS)-stimulated human pulmonary epithelial cell damage. After stimulated by LPS, BEAS-2B cells were treated with various concentrations of Octreotide. CCK-8 assay and LDH kits were to evaluate cell cytotoxicity. ELISA kits were to analyze the levels of inflammatory factors. TUNEL staining was to measure cell apoptosis. Western blot assay was used to assess the expression of apoptosis-related proteins, autophagy-related proteins and AKT/mTOR signaling-related proteins. Then, 3-methyladenine (3-MA) was adopted for treating BEAS-2B cells to determine its effects on inflammation and apoptosis. Afterward, adding AKT agonist (SC79) or mTOR antagonist (rapamycin) to explore the impact of Octreotide on autophagy. Results revealed that Octreotide notably enhanced cell viability and reduced LDH activity. The levels of inflammatory factors were significantly decreased following Octreotide treatment. Additionally, Octreotide attenuated the apoptotic capacity of LPS-induced BEAS-2B cells, led to the up-regulation of Bcl-2 protein level while cut down the protein levels of Bax and cleaved caspase3. Remarkably, the expression of autophagy-related protein LC3II/I and Beclin1 was elevated after Octreotide administration. Importantly, the suppressive effects of Octreotide on the inflammation and apoptosis of LPS-induced BEAS-2B cells was abrogated by 3-MA. Further experiments suggested that Octreotide downregulated p-AKT and mTOR expression in LPS-stimulated BEAS-2B cells. SC79 addition inhibited autophagy, evidenced by downregulated LC3II/I and Beclin1 expression while rapamycin presented the opposite effects. To conclude, Octreotide activates autophagy to alleviate LPS-induced pulmonary epithelial cell injury by inhibiting the AKT/mTOR signaling.
奥曲肽是天然生长抑素的八肽合成物。本研究旨在探讨奥曲肽对脂多糖(LPS)刺激的人肺上皮细胞损伤的影响。用不同浓度的奥曲肽处理 LPS 刺激后的 BEAS-2B 细胞。CCK-8 检测试剂盒和 LDH 试剂盒评估细胞毒性。ELISA 试剂盒分析炎症因子水平。TUNEL 染色法检测细胞凋亡。Western blot 检测凋亡相关蛋白、自噬相关蛋白和 AKT/mTOR 信号通路相关蛋白的表达。然后,用 3-甲基腺嘌呤(3-MA)处理 BEAS-2B 细胞,观察其对炎症和凋亡的影响。接着,加入 AKT 激动剂(SC79)或 mTOR 拮抗剂(雷帕霉素),探讨奥曲肽对自噬的影响。结果表明,奥曲肽显著提高细胞活力,降低 LDH 活性。奥曲肽处理后炎症因子水平显著降低。此外,奥曲肽减轻 LPS 诱导的 BEAS-2B 细胞凋亡能力,上调 Bcl-2 蛋白水平,降低 Bax 和 cleaved caspase3 蛋白水平。奥曲肽给药后自噬相关蛋白 LC3II/I 和 Beclin1 的表达增加。重要的是,3-MA 可阻断奥曲肽对 LPS 诱导的 BEAS-2B 细胞炎症和凋亡的抑制作用。进一步实验表明,奥曲肽下调 LPS 刺激的 BEAS-2B 细胞中 p-AKT 和 mTOR 的表达。SC79 抑制自噬,表现为 LC3II/I 和 Beclin1 表达下调,而雷帕霉素则呈现相反的效果。综上所述,奥曲肽通过抑制 AKT/mTOR 信号通路激活自噬,减轻 LPS 诱导的肺上皮细胞损伤。