The influence of oral copper-methionine on matrix metalloproteinase-2 gene expression and activation in right-sided heart failure induced by cold temperature: A broiler chicken perspective.

This study was designed to investigate the expression, activation and activity of matrix metalloproteinase-2 (MMP-2) in the heart of broiler chickens reared in cold conditions and fed with copper-methionine supplement at different levels. The chickens (n=480) were randomly allotted to six treatments and four replicates. Treatments included two rearing temperatures (i.e. normal and cold temperatures) each combined with three levels of supplemental copper-methionine (i.e. 0, 100 and 200mg/kg). On d 38 and 45 of age, four broilers from each treatment were sacrificed and their hearts were stored at -80°C. Right-sided heart failure, as evident from abdominal and pericardial fluid accumulation, was observed in broilers under cold stress and not receiving supplemental copper. This clinical observation was confirmed at molecular level through increased MMP-2 expression, activation and activity in this group. Birds reared under normal temperature, however, were not involved in right-sided heart failure nor benefitted from copper-methionine supplementation. In contrast, gelatin zymography and real-time PCR demonstrated that dietary supplementation with copper-methionine decreased pro-MMP-2 and MMP-2 in the heart of chickens reared in cold conditions. However, gelatin reverse zymography did not show any difference between treatments in tissue inhibitor of metalloproteinase-2. Level of supplementation showed similar effects on parameters determined. It is concluded that dietary supplementation with copper-methionine reduced right-sided heart failure at clinical and molecular levels in cold-stressed chickens.

Effects of Dietary Copper-Methionine on Matrix Metalloproteinase-2 in the Lungs of Cold-Stressed Broilers as an Animal Model for Pulmonary Hypertension.

The objective of the present study was to investigate the effects of different levels of copper (as supplemental copper-methionine) on ascites incidence and matrix metalloproteinase-2 (MMP-2) changes in the lungs of cold-stressed broilers. For this purpose, 480 1-day-old Ross 308 broiler chickens were randomly assigned to six treatments. Treatments consisted of two ambient temperatures (thermoneutral and cold stress) each combined with 0, 100, and 200 mg supplemental copper/kg as copper-methionine in a 2 × 3 factorial arrangement in a completely randomized design with four replicates. Ascites was diagnosed based on abdominal and pericardial fluid accumulation at 45 days of age. Fourty-eight broilers were killed at 38 and 45 days of age, and their lungs were collected for biological analysis. Results showed that MMP-2 increased in the lungs of ascitic broilers and that copper-methionine supplementation significantly reduced MMP-2 in cold-stressed broiler chickens. Treatments did not affect tissue inhibitor of metalloproteinase-2 (TIMP-2) at 38 and 45 days of age, and no difference was observed between 100 and 200 mg/kg copper-methionine treatments. In conclusion, copper-methionine at higher than conventional levels of supplementation decreased ascites incidence in low temperature through reduced MMP-2 concentration. Further research is warranted to investigate the effect of copper on MMP-2 concentrations in other tissues with high oxygen demand.