Effect of Pilose Antler Peptide on Doxorubicin-induced H9c2 Cells Injury via TGF-β/Smad/ERK Signaling Pathway
Yan Xu1, Jia Zhou1, Guangfu Lv2, Yuexin Liu1, Xintong Zhao1, Xin Li1, Doudan Ye2, Xiaobo Qu1,* and Xiaowei Huang1*
Effects of different concentrations and time points of DOX and PAP on H9c2 cell viability using the CCK-8 assay. (A) DOX treatment concentrations were 0.08, 0.16, 0.32, 0.63, 1.25, 2.5, 5, 10 and 20 μM, and DOX treatment time points were 6, 12, 24 and 48 h, respectively. (B) PAP treatment concentrations were 4.0, 8.0, 16.0, 32.0, 62.5, 125, 250, 500 and 1000 μg/ml, and PAP treatment time points were 24, 48 and 72 h, respectively.
Effect of PAP on H9c2 cells from DOX-induced cell death. PAP treatment concentrations were 4.0, 8.0, 16.0, 32.0, 62.5, 125, 250, 500, 1000 μg/ml for 72 h and DOX treatment with 5 μM for 24 h.
Effects of PAP on DOX-induced LDH leakage and CK-MB level in H9c2 cells. (A) LDH leakage and (B) CK-MB level were detected in the cellular supernatant.
Effects of PAP on DOX-induced Apoptosis and Cell Cycle in H9c2 cells. (A) Apoptosis and (B) Cell Cycle were detected by flow cytometry. (C) Cell Cycle progression and (D) apoptotic rate of H9c2 cells were presented in panel.
Effects of PAP on DOX-induced Bax and Bcl-2 expression in H9c2 cells. Bax and Bcl-2 expression were detected by Immunofluorescence. (A) Fluorescence images of cells showed Bcl-2 immunofluorescence (green), Bax immunofluorescence (red) and DAPI staining (blue). (B) Bar image represents Bax / Bcl-2 expression level.
Effects of PAP on DOX-induced protein level in the TGF-β/SMAD/ERK pathway. The protein levels of (A) TGF-β1, (B) SMAD2, p-SMAD2, (C) SMAD3, p-SMAD3, (D) ERK and p-ERK were detected by western blot. Bar images represent (E) TGF-β2 (F) p-SMAD2/SMAD2, (G) p-SMAD3/SMAD3 and (H) p-ERK/ERK levels, respectively.