β-Caryophyllene as a Potential Protective Agent Against Myocardial Injury: The Role of Toll-Like Receptors

Figures

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  Figure 1. β-Caryophyllene (BCP) structure.

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  Figure 2. Representative pictures of the TTC-stained rat-heart slices showing the effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on myocardial infarction size in ISO-induced myocardial infarction. All values were expressed as mean ± SD (n = 6). ISO: isoproterenol; BCP: β-Caryophyllene; TTC: triphenyl tetrazolium chloride. # indicates statistically significant from normal group, * indicates statistically significant from isoproterenol group, @ indicates statistically significant from β-caryophyllene (100 mg/kg) group (p < 0.05) using one way ANOVA followed by Tukey’s test as a post-hoc analysis.

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  Figure 3. Representative traces of the electrocardiographic (ECG) showing the effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on ECG components in ISO-induced myocardial infarction. All values were expressed as mean ± SD (n = 6). ISO: isoproterenol and BCP: β-Caryophyllene. # indicates statistically significant from normal group, * indicates statistically significant from isoproterenol group, @ indicates statistically significant from β-caryophyllene (100 mg/kg) group (p < 0.05) using one way ANOVA followed by Tukey’s test as a post-hoc analysis.

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  Figure 4. Upper section: effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on Blood Pressure (BP) indices in ISO-induced myocardial infarction: (A) SAP; (B) DAP; (C) MAP; (D) HR. Lower section: shows representative computerized traces of the β-caryophyllene effects on BP. All values were expressed as mean ± SD (n = 6). ISO: isoproterenol; BCP: β-Caryophyllene, SAP: systolic arterial pressure; DAP: diastolic arterial pressure; MAP: mean arterial pressure; and HR: heart rate. # indicates statistically significant from normal group, * indicates statistically significant from ISO group, @ indicates statistically significant from BCP (100 mg/kg) group (p < 0.05) using one-way ANOVA followed by Tukey’s test as a post hoc analysis.

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  Figure 5. Effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on cardiac marker enzymes in ISO-induced myocardial infarction: (A) CPK, (B) CK-MB, (C) LDH, and (D) cTnT. All values were expressed as mean ± SD (n = 6). ISO: isoproterenol; BCP: β-Caryophyllene; CPK: Creatine Phosphokinase; CK-MB: Creatine Kinase-Myocardial Bound; LDH: Lactate dehydrogenase and cTnT: Cardiac Troponin T. # indicates statistically significant from normal group, * indicates statistically significant from isoproterenol group, @ indicates statistically significant from β-caryophyllene (100 mg/kg) group (p < 0.05) using one-way ANOVA followed by Tukey’s test as a post-hoc analysis.

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  Figure 6. Effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on the TLR pathway in ISO-induced myocardial infarction; (A) TLR2 mRNA, (B) TLR4 mRNA, (C) MyD88 mRNA, and (D) TRIF mRNA expression levels. All values were expressed as mean ± SD. ISO: isoproterenol; BCP: β-Caryophyllene and TLR: Toll-like Receptors. # indicates statistically significant from normal group, * indicates statistically significant from isoproterenol group, @ indicates statistically significant from β-caryophyllene (100 mg/kg) group (p < 0.05) using one-way ANOVA followed by Tukey’s test as a post-hoc analysis.

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  Figure 7. Effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on TLR pathway protein content in ISO-induced myocardial infarction: (A) TLR2, (B) TLR4, (C) MyD88, and (D) TRIF protein content. Upper section: representative immunoblots of TLR4, TLR2, MyD88, and TRIF protein content in the normal control heart tissues and in the hearts after isoproterenol-induced myocardial infarction in the absence or presence of pre-treatment with the graded doses of β-caryophyllene. Lower section: Densitometric analysis of TLR4, TLR2, MyD88, and TRIF. Values are mean ± S.E.M (n = 6) for the ratio of TLR4, TLR2, MyD88, and TRIF to β-actin. ISO: isoproterenol and BCP: β-Caryophyllene; # indicates statistically significant from normal group, * indicates statistically significant from isoproterenol group, @ indicates statistically significant from β-caryophyllene (100 mg/kg) group (p < 0.05) using one-way ANOVA followed by Tukey’s test as a post-hoc analysis.

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  Figure 8. Effects of β-caryophyllene treatment (100, 200 mg/kg) for 21 days on HSP-60 and inflammatory markers in ISO-induced myocardial infarction: (A) HSP-60, (B) TNF-α, (C) IL-1β, and (D) NF- κB. All values were expressed as mean ± SD (n = 6). ISO: isoproterenol; BCP: β-Caryophyllene; HSP-60: Heat shock protein 60; TNF-α: Tumor necrosis factor–alpha; IL-Iβ: Interleukin-1β and NFκB: nuclear factor-κB. # indicates statistically significant from normal group, * indicates statistically significant from isoproterenol group, @ indicates statistically significant from the β-caryophyllene (100 mg/kg) group (p < 0.05) using one-way ANOVA followed by Tukey’s test as a post-hoc analysis.

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  Figure 9. Suggested roll of β-caryophyllene on Toll-like receptor signaling. HSP-60: Heat shock protein 60; TLR2: Toll-like receptor 2; MYD88: Myeloid differentiation primary response gene 88; TRIF: TIR-domain-containing adapter-inducing interferon-β; and NFκB: Nuclear Factor-κB.