Activation of p47phox as a Mechanism of Bupivacaine-Induced Burst Production of Reactive Oxygen Species and Neural Toxicity

Figure 3

Membrane translocation of p47phox was increased after bupivacaine exposure, which was inhibited by NOX inhibition. (a, b) The protein level of total p47phox after bupivacaine incubation at different time points was measured by Western blot. The level of p47phox membrane translocation was detected by the ratio of p47phox expressed in the cytosol and membrane. (c, d) SH-SY5Y cells were treated with 1.5 mM bupivacaine for 3 h and incubated with DH10 medium until 24 h. SH-SY5Y cells were pretreated with VAS2870 (10 μM) 30 min prior to bupivacaine treatment. The level of p47phox translocation in SH-SY5Y cells with different treatments. (e, f) The level of p47phox translocation in animal models. (g) The level of p47phox membrane translocation was detected by immunofluorescent staining. Pan-cadherin is a widely used membrane marker and was stained green, targeted protein p47phox was stained red, and nucleus was stained with DAPI (blue). Data represent mean ± SD of at least 3 independent experiments or for 6 rats each group. Scale bar: 10 μm. versus Con group. ^# versus Bup group. ^& versus VAS group.

(a)

(b)

(c)

(d)

(e)

(f)

(g)