Liu Chuanlan, Zhu Xiaomu, Aga Erbu, Tse Wai Ming, Tse Kathy Wai Gaun, Liu Yanyong, Ye Bengui
Medical College of Tibet University, Lasa 850002, China; Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry, Sichuan Engineering Laboratory for Plant-Sourced Drug, and Sichuan Research Center for Drug Precision Industrial Technology, West China School of Pharmacy, Sichuan University, Chengdu 610041, China.
Department of Pharmacy, West China Hospital of Sichuan University, Chengdu 610041, Sichuan, China.
Cell Stress Chaperones. 2024 Dec;29(6):697-708. doi: 10.1016/j.cstres.2024.10.001. Epub 2024 Oct 4.
Ebeiedinone and peimisine are the major active ingredients of Fritillariae Cirrhosae Bulbus. In this study, we looked at how these two forms of isosteroidal alkaloids protect human bronchial epithelial BEAS-2B cells from oxidative stress and apoptosis caused by cigarette smoke extract (CSE). First, the cytotoxicity was determined using the CCK8 assay, and an oxidative stress model was established. Then the antioxidative stress activity and mechanism were investigated by ELISA, flow cytometry, and Western blotting. By the CCK-8 assay, exposure to CSE (20%, 40%, and 100%) reduced the viability of BEAB-2S cells. The flow cytometry findings indicated that CSE-induced production of ROS (0.5% to maximum) and treatments with 10 μM ebeiedinone and 20 μM peimisine attenuated the production of ROS. The western blot assay results indicate that ebeiedinone and peimisine reduce CSE-induced oxidative stress, DNA damage, apoptosis, and autophagy dysregulation by inhibiting ROS, upregulating SOD and GSH/GSSG, and downregulating MDA, 4-HNE, and 8-OHdG through the NRF2/KEAP1 and JNK/MAPK-dependent pathways, thereby delaying the pathological progression of COPD caused by CS.Our data suggest that CSE causes oxidative stress, DNA damage, and apoptosis in BEAS-2B cells, as well as the progression of COPD. Ebeiedinone and peimisine fight CS-induced COPD by suppressing autophagy deregulation and apoptosis.
浙贝乙素和浙贝母碱是川贝母的主要活性成分。在本研究中，我们观察了这两种异甾体生物碱如何保护人支气管上皮BEAS-2B细胞免受香烟烟雾提取物（CSE）引起的氧化应激和细胞凋亡。首先，使用CCK8测定法确定细胞毒性，并建立氧化应激模型。然后通过ELISA、流式细胞术和蛋白质印迹法研究抗氧化应激活性及其机制。通过CCK-8测定法，暴露于CSE（20%、40%和100%）会降低BEAB-2S细胞的活力。流式细胞术结果表明，CSE诱导ROS产生（0.5%至最大值），而用10μM浙贝乙素和20μM浙贝母碱处理可减弱ROS的产生。蛋白质印迹分析结果表明，浙贝乙素和浙贝母碱通过抑制ROS、上调SOD和GSH/GSSG以及通过NRF2/KEAP1和JNK/MAPK依赖性途径下调MDA、4-HNE和8-OHdG，从而减少CSE诱导的氧化应激、DNA损伤、细胞凋亡和自噬失调，进而延缓由香烟烟雾引起的慢性阻塞性肺疾病（COPD）的病理进展。我们的数据表明，CSE会导致BEAS-2B细胞发生氧化应激、DNA损伤和细胞凋亡，以及COPD的进展。浙贝乙素和浙贝母碱通过抑制自噬失调和细胞凋亡来对抗香烟烟雾诱导的COPD。
