| Objective:Crocetin is one of the major active constituents of saffron extract, and it is a carotenoid compound that prevents reactive oxygen species and has anti-inflammation and anti-apoptosis characteristics. This study aims to investigate whether crocetin repairs myocardial damage in vivo after ischemia reperfusion (I/R) and the mechanisms underlying its cardioprotective effects.Methods:Male wistar rats were randomly allocated into three groups:Sham, I/R, CRO. Crocetin (50 mg/kg/day, i.g.) or sodium carboxymethylcellulose (CMC-Na) were intragastrically administered to Wistar rats for 7 d before operation. Myocardial ischemia reperfusion injury (MIRI) was induced by occluding the left anterior descending (LAD) coronary artery for 45 min and subsequent reperfusion for 3 h. After the operation,we used 2,3,5-triphenyl four azole nitrogen chloride (TTC) staining to measured the myocardial infarction size; hematoxylin-eosin (HE) staining to evaluation the myocardial Histopathological change; Xanthine oxidase method to detect serum total superoxide dismutase (T-SOD) activity and thiobarbituric acid method to detect serum malondialdehyde (MDA) content; Enzyme-linked immunosorbent assay (ELISA) staining to detective the serum concentration of Creatine Kinase Isoenzyme-MB (CK-MB), Tumor Necrosis Factor-a (TNF-a) and Interleukin-10 (IL-10); Immunohistochemistry to detect Bcl-2 or Bax protein of intracellular content and Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) to detect the mRNA expression of Bcl-2 and Bax; terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining to measure the myocardial apoptosis index.Results:(1) After reperfusion, compared with Sham group, ischemic myocardial fiber disarray, fracture, widened interstitial edema, myocardial fibers seen a lot of inflammatory cell infiltration, and the drug group, myocardial fiber disarray, visible necrosis foci, but clearer than ischemia group.(2) Compared to Sham group, I/R group and CRO group serum CK-MB, MDA, TNF-a levels increased significantly (P<0.01), and T-SOD levels decreased significantly (P<0.01), the levels of IL-10 increased significantly in CRO group (P<0.01); Compared to I/R group, serum CK-MB, TNF-a, MDA levels and myocardial apoptosis index in CRO group was significantly lower (P<0.01), whereas serum IL-10 and T-SOD levels were significantly increased (P<0.01).(3) Compared to Sham group, the area of myocardial ischemia infarction, apoptosis index, Bax protein expression are decreased (P<0.01), and Bax mRNA expression is also decreased (P<0.05); while Bcl-2 protein and mRNA expression are increased significantly (P<0.01).Conclusion:(1) Crocetin pretreatment have protective effects on myocardial ischemia reperfusion injury in rats, the mechanism of it may be through inhibit the production of reactive oxygen species and inflammatory reactive, reducing myocardial apoptosis.(2) Crocetin pretreatment can inhibit the myocardial apoptosis of ischemia reperfusion injury rats, the mechanism of it may be up-regulated apoptotic inhibitory protein Bcl-2 expression and down-regulated pro-apoptotic protein Bax expression, eventually up regulated the ratio of Bcl-2/Bax. |
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