Cardioprotection of tilianin ameliorates myocardial ischemia-reperfusion injury: Role of the apoptotic signaling pathway

Figures

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  Fig 1. (A): Chemical structure of tilianin. Molecular weight: 446.4. Molecular formula: C22H22O10. (B): HPLC chromatogram of tilianin control. (C): HPLC chromatogram of tilianin sample.

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  Fig 2. Effect of pre-administration with tilianin on ST-segment elevation after MIRI in rats. Rats were subjected to 45 min of ischemia followed 4 h of reperfusion was measured. (Effect of tilianin on ST-segment elevation. Results are expressed as mean ± SD. The traces of ECG in various groups, a: Sham; b: MIRI; c: L-tilianin (2.5 mg/kg); d: M-tilianin (5 mg/kg) and e: H-tilianin (10 mg/kg). ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10).

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  Fig 3. Images of TTC and Evans Blue stained heart sections, and the quantitative analysis. (a: Sham; b: MIRI; c: L-tilianin; d: M-tilianin; e: H-tilianin, #P<0.05, ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10).

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  Fig 4. Echocardiographic analysis. (a: Sham; b: MIRI; c: L-tilianin; d: M-tilianin; e: H-tilianin, #P<0.05, ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10).

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  Fig 5. Detection apoptotic myocytes by TUNEL assay in each group. Representative images from Sham group, MIRI group and various administrations are shown at 400× magnifications. (a: Sham; b: MIRI; c: L-tilianin; d: M-tilianin and e: H-tilianin, #P<0.05, ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10).

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  Fig 6. (A): Bcl-2, Bax and GAPDH expression levels in the myocardial tissue of each group. (1: Sham, 2: MIRI, 3: L-tilianin, 4: M-tilianin and 5: H-tilianin) (#P<0.05, ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10). (B): Cleaved-caspase-3, caspase-3 and β-actin expression levels in the myocardial tissue of each group. (1: Sham, 2: MIRI, 3: L-tilianin, 4: M-tilianin and 5: H-tilianin) (#P<0.05, ##P<0.01 vs. the Sham group; **P<0.01 vs. the MIRI group, n = 10). (C): XIAP, HtrA2/Omi, Smac/Diablo and β-actin expression levels in the myocardial tissue of each group. (1: Sham, 2: MIRI, 3: L-tilianin, 4: M-tilianin and 5: H-tilianin) (#P<0.05, ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10). (D): Caspase-7, Caspase-9 and β-actin expression levels in the myocardial tissue of each group. (1: Sham, 2: MIRI, 3: L-tilianin, 4: M-tilianin and 5: H-tilianin) (#P<0.05, ##P<0.01 vs. the Sham group;*P<0.05, **P<0.01 vs. the MIRI group, n = 10).

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  Fig 7. (A): p-Akt, Akt and GAPDH expression levels in in the myocardial tissue of each group. (1: Sham, 2: MIRI, 3: L-tilianin, 4: M-tilianin, 5: H-tilianin) (##P<0.01, **P<0.01, n = 10). (B): p-PI3K, PI3K and GAPDH expression levels in in the myocardial tissue of each group. (1: Sham, 2: MIRI, 3: L-tilianin, 4: M-tilianin, 5: H-tilianin) (##P<0.01 vs. the Sham group; **P<0.01 vs. the MIRI group, n = 10).

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  Fig 8. (A): p-Akt, XIAP, Akt and GAPDH expression levels in the myocardial tissue of each group. (a: MIRI+H-tilianin and b: MIRI+H-tilianin+LY294002) (**P<0.01 vs. the MIRI+H-tilianin+LY294002 group, n = 10). (B): Echocardiographic analysis (a: MIRI+H-tilianin and b: MIRI+H-tilianin+LY294002). (**P<0.01 vs. the MIRI+H-tilianin+LY294002 group, n = 10).

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  Fig 9. Putative mechanism of tilianin improved myocardial apoptosis. (Tilianin pretreatment significantly activated PI3K/Akt pathway and inhibited release of Smac/Diablo and HrtA2/Omi from mitochondria, XIAP degradation and caspase activation).