Statin rosuvastatin inhibits apoptosis of human coronary artery endothelial cells through upregulation of the JAK2/STAT3 signaling pathway

Abstract

The purpose of the present study was to explore the potential molecular signaling pathway mediated by the statin rosuvastatin in cultured human coronary artery endothelial cells (HCAECs) induced by CoCl2. CoCl2 was used to induce the apoptosis of HCAECs. Myocardial infarction rats were established and received statin or PBS treatment. Reverse transcription‑quantitative PCR, western blotting, ELISA, TUNEL assay and immunohistochemistry were used to analyze the role of statin treatment. The results showed that rosuvastatin treatment decreased apoptosis of HCAECs induced by CoCl2 by increasing anti‑apoptosis Bcl‑xl and Bcl‑2 expression, and decreasing pro‑apoptosis Bax, Bad, caspase‑3 and caspase‑9 expression. The myocardial ischemia rat model demonstrated that rosuvastatin treatment decreased the mitochondrial reactive oxygen species, inflammation, mitochondrial damage, lipid catabolism, heart failure and the myocardial infarction areas, but improved the cardiac function indicators, right and left ventricular ejection fraction and increased expression levels of Janus kinase (JAK) and signal transducer and activator of transcription (STAT)3 in myocardial tissue. In conclusion, the results of the current study revealed that the statin rosuvastatin presents cardioprotective effects by activation of the JAK2/STAT3 signaling pathway.

Keywords: myocardial infarction; statin; rosuvastatin; apoptosis; human coronary artery endothelial cells; Janus kinase 2; signal transducer and activator of transcription 3.

Figure 1.

Statin rosuvastatin increases the viability of HCAECs induced by CoCl₂. (A) CoCl₂ decreased viability of HCAECs in a dose-dependent manner (0, 10, 20 and 30 mM) compared with the non-treated group. (B) CoCl2 decreased viability of HCAECs in a time-dependent manner (24, 48 and 72 h) compared with the non-treated group. (C) Rosuvastatin increased the viability of HCAECs in dose-dependent manner (0, 1.0, 2.0 and 3.0 mg/ml) compared with the non-treated group. (D) Statin increased viability of HCAECs in a time-dependent manner (24, 48 and 72 h) compared with non-treated group. *P<0.05, **P<0.01; ns, not significant; HCAECs, human coronary artery endothelial cells.

Figure 2.

Statin rosuvastatin inhibits apoptosis of HCAECs induced by CoCl₂. (A) Statin inhibited apoptosis of HCAECs induced by CoCl₂ (magnification, ×50). (B and C) Statin treatment increased anti-apoptosis Bcl-xl and Bcl-2 mRNA (B) and protein (C) expression in the HCAECs. (D and E) Statin treatment decreased Bax and Bad mRNA (D) and protein (E) in the HCAECs. (F and G) Statin treatment decreased apoptotic markers caspase-3 and caspase-9 mRNA (F) and protein (G) in the HCAECs. **P<0.01, compared to the Control group. HCAECs, human coronary artery endothelial cells.

Figure 3.

Statin rosuvastatin suppresses the apoptosis of HCAECs through regulation of the JAK2/STAT3 pathway. (A) JAK2 selective inhibitor AG490 decreased and abolished statin-regulated JAK2, p-JAK2, STAT3 and p-STAT3 expression in the HCAECs. (B) JAK2 selective inhibitor AG490 decreased and abolished statin-inhibited apoptosis of HCAECs (magnification, ×50). (C) Effects of JAK2 selective inhibitor AG490 on statin-regulated Bcl-xl and Bcl-2 protein expression in the HCAECs. (D) Effects of JAK2 selective inhibitor AG490 on statin-regulated Bax and Bad protein expression in the HCAECs. (E) Effects of JAK2 selective inhibitor AG490 on statin-regulated Bcl-xl and Bcl-2 gene expression in the HCAECs. (F) Effects of JAK2 selective inhibitor AG490 on statin-regulated Bax and Bad gene expression in HCAECs. **P<0.01; ns, not significant; HCAEC, human coronary artery endothelial cells; JAK, Janus kinase; p-, phosphorylated; STAT, signal transducer and activator of transcription.

Figure 4.

Effects of statin rosuvastatin on inflammatory markers and cellular parameters in a myocardial infarction rat model. Effects of statin on (A) blood hsCRP concentration, (B) number of WBCs, (C) LDL-c, (D) neutrophils, (E) MPV, (F) LVEF and (G) LVFS in myocardial infarction rat model. *P<0.05, **P<0.01. hsCRP, hypersensitive C-reactive protein; WBC, leucocytes; LDL-c, low-density lipoprotein cholesterol; MPV, mean platelet volume; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening.

Figure 5.

Expression levels of inflammatory factors, mitochondrial damage and cardiac hypertrophic failure in a myocardial infarction rat model. (A) Effects of statin rosuvastatin on the protein levels of MMP-9, TNF-α, NF-κB and IL-1β in heart tissue. (B) Effects of statin on anti-inflammatory cytokine IL-10 in heart tissue. (C) Effects of statin on cytosolic Cytochrome c expression in heart tissue. (D) Effects of statin on PGC-1α, BNP and α-MHC expression in heart tissue. **P<0.01. MMP, matrix metalloproteinase; TNF, tumor necrosis factor; NF-κΒ, nuclear factor κΒ; IL, interleukin; PGC, peroxisome proliferator-activated receptor γ coactivator; BNP, brain natriuretic peptide; α-MHC, α myosin heavy chain.

Figure 6.

In vivo efficacy of statin rosuvastatin on myocardial infarction. (A) Statin treatment markedly decreased the mitochondrial ROS in a myocardial infarction rat model. (B) Statin treatment decreased myocardial infarction area in the experimental rats. (magnification, ×50). (C) Statin treatment decreased thrombogenesis in the experimental rats after the 60-day treatment. (D) Statin treatment decreased Bax and Bad production in the experimental rats after the 60-day treatment. (E) Statin improved the cardiac function indicators LVPWd and LVEDD. (F) Statin increased expression level of JAK and STAT3 in myocardial tissue. Scale bar, 50 µm. *P<0.05, **P<0.01. ROS, reactive oxygen species; LVPWd, left ventricular end-diastolic posterior wall thickness; LVEDD, left ventricular end-diastolic diameter; JAK, Janus kinase; STAT, signal transducer and activator of transcription.