Necrostatin-1 protects C2C12 myotubes from CoCl2-induced hypoxia.

摘要: Necrostatin-1 (Nec-1) is a selective and potent allosteric inhibitor of necroptosis by specifically inhibiting the activity receptor‑interacting protein (RIP) 1 kinase. The aim present study was to determine effect Nec‑1 on an anoxia model comprising mouse skeletal C2C12 myotubes. In study, hypoxic mimetic reagent, cobalt chloride (CoCl2), used induce hypoxia in cytotoxic effects CoCl2‑induced were determined Cell Counting kit‑8 assay flow cytometry. Transmission electron microscopy (TEM) characterize morphological characteristics dead cells at ultrastructural level. To clarify signaling pathways CoCl2‑mediated cell death, expression levels RIP1, RIP3, extracellular signal‑regulated kinase (ERK)1/2, hypoxia‑inducible factor (HIF)‑1α B lymphoma‑2 adenovirus E1B 19‑kDa interacting 3 (BNIP3) investigated western blotting. Oxidative stress using 2',7'‑dichlorofluorescin diacetate measure intracellular reactive oxygen species (ROS) fluorescent dye JC‑1 mitochondrial membrane potential (Δψm). results showed that ratios apoptotic necrotic increased following CoCl2 treatment, typical necroptotic able observe TEM, whereas exhibited protective against oxidative stress. Treatment with significantly decreased p‑ERK1/2, HIF‑1α, BNIP3 ROS induced CoCl2, promoted differentiation. reversed decrease potential. Together, these findings suggested protected myotubes under conditions CoCl2-induced hypoxia.