Butorphanol protects PC12 cells against OGD/R-induced inflammation and apoptosis

Abstract

The aim of the present study was to examine the effects of butorphanol on neural injury in an oxygen glucose deprivation/reoxygenation (oGd/r) model using Pc12 cells, and to investigate whether mitochondrial apoptosis was involved in these effects. To establish the oGd/r model, Pc12 cells were cultured under hypoxia and low glucose conditions. Expression levels of inflammatory cytokines were evaluated by detecting the levels of tumor necrosis factor-α, interleukin (IL)-1β, il-6 and monocyte chemoattractant protein-1. oxidative stress was evaluated by measuring the levels of reactive oxygen species, lactate dehydrogenase activity and myeloperoxidase concentration. apoptosis, protein expression and cell viability were determined by flow cytometry, western blotting and by using a cell counting Kit-8, respectively. compared with the control group, cell viability, expression of inflammatory factors and oxidative stress were all decreased in the oGd/r group. all the above changes could be mitigated by treatment with butorphanol. in addition, butorphanol treatment resulted in a significant upregulation of Bax, and downregulation of Bcl-2, activated caspase-3, caspase-9 and poly adP-ribose polymerase, increased the expression of X-linked inhibitor of apoptosis protein and enhanced ATP activity. To conclude, these results suggested that the protective effects of butorphanol are associated with the inhibition of OGD/R-induced inflammation and apoptosis injury, and may be partially associated with the inhibition of mitochondrial apoptosis.

Butorphanol reduces the neuronal inflammatory response and apoptosis via inhibition of p38/JNK/ATF2/p53 signaling

“…5). A previous study has indicated that butorphanol can relieve neuronal inflammation and apoptosis arising from ischemia-hypoxia-reperfusion (14) and the present study is consistent with the previous studies, which all indicated that neuronal inflammation and apoptosis are alleviated following butorphanol treatment. The novelty of the present study was that it explored the protective effects on LPS-induced PC12 cells through p38/JNK/ATF2/p53 signaling.…”

“…Cells were divided into the following groups: Untreated (control), butor phanol, lipopolysaccha r ide (LPS), LPS + butorphanol and LPS + butorphanol + anisomycin. Cells were pretreated with butorphanol (1, 2 and 4 µM; GlpBio Technology) (14) and/or anisomycin (1 µM; GlpBio Technology Inc.) (22) for 6 h and then induced with LPS (5 µg/ml; MilliporeSigma) (23) for 12 h for the establishment of the model.…”

“…PC12 cells were seeded in a 96-well plate at a density of 6x10 3 cells/well. To determine the effect of different doses of butorphanol on cell viability, butorphanol (1, 2 and 4 µM) was added to the plates and cells were incubated for 24 h as in a previous study (14). To determine the effect of LPS and butorphanol on cell viability, cells were pretreated with butorphanol for 6 h and then incubated with LPS for 12 h. Following incubation with 10 µl CCK8 solution (Beyotime Institute of Biotechnology) for another 2 h at 37˚C, the optical density was measured at a wavelength of 450 nm using a microplate reader (Thermo Fisher Scientific, Inc.).…”

“…In addition, butorphanol can reduce inflammatory infiltration injury to alleviate brain damage as a consequence of sepsis (13). Furthermore, butorphanol is considered to relieve neuronal inflammation and apoptosis arising from ischemia-hypoxia-reperfusion (14). It was hypothesized that butorphanol may protect against inflammation and apoptosis in neuronal injury.…”

Exploration of the mechanism of luteolin against ischemic stroke based on network pharmacology, molecular docking and experimental verification

“…The membranes were then blocked with 5% skim milk (Absin, Shanghai, China) for 2 h at room temperature and followed by incubation overnight at 4°C with primary antibodies directed against MMP9(sc-393,859, Santa, USA), MAPK3(sc-271,269, Santa, USA), PTGS2(sc-376,861, Santa, USA), JUN (#9165, CST, USA), IL1B(sc-12,742, Santa, USA) and TNF (#11,948, CST, USA). After washing with TBST for 30 minutes at room temperature, we incubated the immune blots with secondary antibodies(#7074, CST, USA) for 2 h. Finally, proteins were visualized by Yeasen super ECL detection reagent Kit (Yeasen, China) using Syngene Bio Imaging instrument (Synoptics, Cambridge) and were quantified by Image J [ 42 ].…”