Chlorpromazine and Promethazine (C+P) Reduce Brain Injury after Ischemic Stroke through the PKC-δ/NOX/MnSOD Pathway

Figure 6

C+P suppressed the translocation of PKC-δ and interaction of PKC-δ with p47^phox induced by OGD/R in SHSY5Y cells. (a) Representative bands of cytosolic and membrane PKC-δ identified using Western blot at 6 and 24 hours of reperfusion. Bar graphs show semiquantitative PKC-δ levels in the cytosol and membrane determined using band density analysis. Cytosolic PKC-δ expression decreased but increased in the membrane in the OGD/R group at 6 hours after reoxygenation as compared to the control group. Administration of C+P negated the changes of OGD/R-induced PKC-δ expression. No differences were detected in cytosolic PKC-δ expression in the OGD/R group at 24 hours of reoxygenation, but OGD/R increased membranous PKC-δ expression as compared to the control group. C+P attenuated the raised cellular membrane PKC-δ concentration. , as compared to the OGD group; ^#, ^##, as compared to the OGD/C+P group. All the quantifications were normalized to α-tubulin (cytosol) or Na⁺/K⁺ ATPase (membrane), respectively. Results are shown as of three independent experiments. (b) Representative bands of coimmunoprecipitation of PKC-δ and p47^phox in SHSY5Y cells, identified via the Western blot technique at 6 and 24 hours of reoxygenation. Bar graphs show the ratios of p47^phox/PKC-δ identified using band density analysis. IgG antibody served as the control. In comparison to IgG control, a significant enhancement in p47^phox was detected in the immunoprecipitation complex with the PKC-δ antibody from cell lysates, which suggests a heightened interaction of PKC-δ and p47^phox. Moreover, the interaction of PKC-δ and p47^phox increases after 6 hours of reoxygenation, while C+P treatment reduced the interaction. Although the interaction between the control and the OGD group was not significant at 24 hours of reoxygenation, the interaction did increase, and C+P treatment lowered the interaction between PKC-δ and p47^phox. versus the OGD group; ^#, ^## versus the OGD/C+P group. The data is represented as of three independent experiments.

(a)

(b)