PKA Inhibitor H89 (N-[2-p-bromocinnamylamino-ethyl]-5-isoquinolinesulfonamide) Attenuates Synaptic Dysfunction and Neuronal Cell Death following Ischemic Injury

Figure 5

The measurement of cleaved caspase-3 and Bcl2 expression in Neuro2A cells after H/R-induced injury. (a) The level of cleaved caspase-3 was evaluated by immunocytochemistry. This image shows that the expression of cleaved caspase-3 in the H/R group was strongly increased compared to the normal group. Cleaved caspase-3 expression was attenuated in H89 pretreatment treatment group under H/R-induced injury. (b) The level of Bcl2 was evaluated by immunocytochemistry. This image shows that the expression of Bcl2 in the H/R group was increased compared to the normal group. PKA inhibitor H89 pretreatment preserved the expression of Bcl2 in spite of hypoxia reperfusion injury. (c) Western blotting experiments showed that the relative protein expression of cleaved caspase-3 evidently attenuated in the H89 group compared to the hypoxia reperfusion group. (d) Western blotting experiments showed that the relative protein expression of Bcl2 slightly increased in the H89 group compared to the hypoxia reperfusion group. Data were expressed as mean ± S.E.M, and each experiment included 4 repeats per condition. Differences were considered significant . Scale bar: 200 μm, cleaved caspase-3: red, Bcl2: green, 4′,6-diamidino-2-phenylindole (DAPI): blue, normal: the normal control group, H/R: 4 hr hypoxia and 18 hr reperfusion injury group, and H89: 2 hr PKA inhibitor H89 treatment group before 4 hr hypoxia and 18 hr reperfusion injury.

(a) Cleaved caspase-3

(b) Bcl2

(c) Cleaved caspase-3

(d) Bcl2