Angiotensin II upregulates cyclophilin A by enhancing ROS production in rat cardiomyocytes

Figures

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  Figure 1.. Ang II upregulates CypA in rat cardiomyocytes. (A) Western blotting and (B) semi-quantification analysis demonstrated that CypA expression was stimulated by Ang II in cultured H9C2 cells. (C) Ang II increased CypA content in the supernatant of cultured H9C2 cells. n=3/group. *P<0.05. Ang II, angiotensin II; CypA, cyclophilin A.

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  Figure 2.. Ang II upregulates CypA via ROS. (A) Representative images and (B) analysis demonstrating that Ang II increased ROS production in cultured H9C2 cells. n=3/group. Green fluorescence represents ROS; scale bar, 50 µm. (C) Western blotting and (D) semi-quantification analysis demonstrated that Ang II-induced CypA expression was inhibited by GEE, a ROS inhibitor. *P<0.05. Ang II, angiotensin II; CypA, cyclophilin A; GEE, glutathione ethyl ester; ROS, reactive oxygen species.

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  Figure 3.. Elevated ROS induce the expression of CypA. (A) Representative images and (B) analysis demonstrating that butyrate promoted ROS production in cultured H9C2 cells. n=3/group; scale bar, 50 µm. (C) Western blotting and (D) semi-quantification analysis of CypA expression in H9C2 cells following treatment with butyrate. (E) CypA content analysis in the supernatant of H9C2 cells following treatment with butyrate. *P<0.05. CypA, cyclophilin A; ROS, reactive oxygen species.

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  Figure 4.. Ang II enhances ROS production via AT2R. (A) Representative images and (B) analysis of ROS production in Ang II-induced H9C2 cells pretreated with Valsartan (angiotensin receptor 1 antagonist) and PD123319 (AT2R antagonist). Scale bar, 50 µm. PD123319 significantly inhibited Ang II-induced ROS production. n=3/group. (C) Western blotting and (D) semi-quantification analysis of CypA expression in Ang II-induced H9C2 cells pretreated with Valsartan and PD123319. PD123319 significantly suppressed Ang II-induced CypA expression in H9C2 cells. n=3/group. *P<0.05, ***P<0.01. Ang II, angiotensin II; AT2R, angiotensin receptor 2; CypA, cyclophilin A; ROS, reactive oxygen species.

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  Figure 5.. Proposed mechanism of how Ang II upregulates CypA. Ang II increased ROS levels via AT2R in cardiomyocytes; increased ROS, in turn, promoted CypA expression and secretion. Ang II, angiotensin II; AT2R, angiotensin type 2 receptor; CypA, cyclophilin A; ROS, reactive oxygen species.