Department of Anesthesiology, Tongzhou Maternal and Child Health Hospital of Beijing, Beijing, China.
Department of General Medicine, Beijing Luhe Hospital, Capital Medical University, Beijing, China.
Histol Histopathol. 2021 Apr;36(4):425-435. doi: 10.14670/HH-18-298. Epub 2021 Jan 7.
Propofol (PRO) was reported to exert a neuroprotective effect by decreasing microRNA-134 (miR-134), a brain-specific miRNA, thus, the role of PRO against cobalt chloride (CoCl₂)-induced injury in rat pheochromocytoma cells (PC12) via mediating miR-134 was explored.
CoCl₂-induced PC12 cells treated with PRO were transfected with or without miR-134 negative control (NC)/ inhibitor/mimic, and the following detections were then performed using cell counting kit-8 (CCK-8), Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) and Hoechst 33258 staining. Autophagy was observed by transmission electron microscope (TEM). Mitochondrial membrane potential (MMP) was detected by Rhodamine-123 (Rh123) staining, and reactive oxygen species (ROS) by dichloro-dihydro-fluorescein diacetate (DCFH-DA) staining. Protein and gene expressions were measured by Western blotting and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), respectively.
PRO reversed the CoCl₂-induced decrease in the PC12 cell viability and increased miR-134 in a dose-dependent manner. CoCl₂ increased LC3II/I ratio and Beclin-1 expression, but decreased p62 expression, which was abolished by PRO. In addition, an increased cell apoptosis rates triggered by CoCl₂ were reduced by PRO with the down-regulations of Bax and Caspase-3 and the up-regulation of Bcl-2. Furthermore, PRO decreased methylenedioxyamphetamine (MDA), nitric oxide (NO) and ROS in CoCl₂-induced PC12 cells accompanying the increase in glutathione peroxidase (GSH-Px) and MMP. The effects of PRO on autophagy, apoptosis and oxidative stress in CoCl₂-induced PC12 cell were reversed by miR-134 mimic.
PRO may mitigate CoCl₂-induced autophagy in PC12 cells with decreased apoptosis and improved oxidative stress via mediating miR-134.
据报道，异丙酚（PRO）通过降低脑特异性 microRNA-134（miR-134）发挥神经保护作用，因此，通过调节 miR-134，探索 PRO 对氯化钴（CoCl₂）诱导的大鼠嗜铬细胞瘤细胞（PC12）损伤的作用。
用 PRO 处理 CoCl₂诱导的 PC12 细胞，转染 miR-134 阴性对照（NC）/抑制剂/模拟物，然后使用细胞计数试剂盒-8（CCK-8）、Annexin V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）和 Hoechst 33258 染色进行以下检测。通过透射电子显微镜（TEM）观察自噬。通过 Rhodamine-123（Rh123）染色检测线粒体膜电位（MMP），通过二氯二氢荧光素二乙酸酯（DCFH-DA）染色检测活性氧（ROS）。通过 Western blot 和定量逆转录聚合酶链反应（qRT-PCR）分别测量蛋白质和基因表达。
PRO 逆转了 CoCl₂诱导的 PC12 细胞活力下降，并呈剂量依赖性增加 miR-134。CoCl₂增加 LC3II/I 比值和 Beclin-1 表达，但降低 p62 表达，PRO 可消除这一作用。此外，PRO 还降低了 CoCl₂诱导的细胞凋亡率，下调 Bax 和 Caspase-3，上调 Bcl-2。此外，PRO 降低了 CoCl₂诱导的 PC12 细胞中甲基乙二醛（MDA）、一氧化氮（NO）和 ROS，同时增加了谷胱甘肽过氧化物酶（GSH-Px）和 MMP。PRO 对 CoCl₂诱导的 PC12 细胞自噬、凋亡和氧化应激的影响被 miR-134 模拟物逆转。
PRO 可能通过调节 miR-134 减轻 CoCl₂诱导的 PC12 细胞自噬，减少细胞凋亡，改善氧化应激。
