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. 2021 Feb 28;13(3):311. doi: 10.3390/pharmaceutics13030311

Figure 5.

Figure 5

JP4-039 rescues PX-induced toxicity and permeability increase in the BBB injury model; (A) A chemical formula diagram of JP4-039; (B) Permeability of sodium fluorescein (NaF) was measured across the BBB in vitro model (from luminal to abluminal side) 24-h post PX treatment. PX was added at 900 µM and JP4-039 was added at 25 µM. Ctrl are cells cultured with normal medium without the addition of any treatments. Mean ± s.d. of one representative experiment is shown, out of four independent experiments with n = 12–20, *** p < 0.001 vs. ctrl and # p < 0.05, vs. PX; (C) Using the same model, TEER was also measured. Mean ± s.d. of one representative experiment is shown, out of four independent experiments with n = 12–20, *** p < 0.001 vs. ctrl and ## p < 0.01 vs. PX; (D) Cell death was also assessed in the same model by the release of lactate dehydrogenase (LDH).Mean ± s.d. of one representative experiment is shown, out of three independent experiments with n = 9–18, *** p < 0.001 vs. ctrl and ### p < 0.001 vs. PX; (E) Live cell imaging of cell death was examined by cytotoxicity fluorescent staining 24-h post 400 µM PX treatment on a BLEC monolayer. JP4-039 was added at 624 nM. Mean ± s.d. of one representative experiment is shown, out of four independent experiments with n = 16–24, *** p < 0.001 vs. ctrl and # p < 0.05 vs. PX marks the first time point with a significant change; (F) Apoptosis was measured with a caspase-3 activated fluorescent marker by live cell imaging 24-h post PX treatment on a BLEC monolayer. Mean ± s.d. of one representative experiment is shown, out of four independent experiments with n = 16–24, * p < 0.05 vs. ctrl and ## p < 0.01 vs. PX marks the first time point with a significant change; (G) Live cell imaging of mitochondrial ROS formation was measured with Mitosox staining on a BLEC monolayer treated with PX for 24 h. Mean ± s.d. of one representative experiment is shown, out of five independent experiments with n = 20–30, * p < 0. 05 vs. ctrl and # p < 0.05 vs. PX marks the first time point with a significant change. Ctrl, control; PX, paraoxon; TEER, Trans endothelial electrical resistance; MRF, mitochondrial ROS formation; au, arbitrary units.