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Purpose of peroxisome proliferator-activated receptors (peroxisome proliferator-activated receptor, PPAR)-α is a ligand-activated nuclear receptor family of three subtypes of PPARs. Wy14643 are PPAR-α agonist, its protective effect on the important organs and PPARα agonists may have anti-inflammatory and antioxidant effects associated. In the liver, liver cells overexpression of PPAR-α, and no expression of liver Kupffer cells. Our previous studies have shown that, Wy14643 part in donor liver ischemia / reperfusion injury in rats, but different doses Wy14643 on primary hepatocyte hypoxia / reoxygenation injury has not been reported, we study different doses Wy14643 on primary rat hepatocytes hypoxia / reoxygenation injury in rats and its mechanism. Methods 50 male SD rats, weighing 250-300g, taken through a step perfusion primary hepatocytes. The isolated hepatocytes cultured species at 1 × 105/ml in culture plates, each with 6 holes. Randomly divided into six groups: normal group (C), hypoxia group (H / R), DMSO group (D), Wy14643 (10,30,100 umol / L) group. Wy14643 group, hypoxia / reoxygenation 2 hours before the Wy14643 added to the medium, the culture medium was adjusted to a final concentration Wy14643 10,30,100 umol / L. The liver cells into the incubator, hypoxia (37 ℃, 95% N2-5% CO2) 4 hours reoxygenation (37 ℃, 95% O -5% CO2) 10 小时 induced cell damage, and then were collection of liver cells and cell culture supernatants were measured liver cell culture supernatant alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity and liver malondialdehyde (MDA) content and superoxide dismutase (SOD) activity; liver mitochondrial glutathione (GSH) levels; MTT method for the determination of liver cell survival; electron microscopy of liver cell damage; RT-PCR detection PPARαmRNA expression. Statistical analysis: using SPSS11.5 software for statistical analysis, measurement data were expressed as mean ± standard deviation (± s) that the groups were compared using one-way ANOVA, P lt; 0.05 was considered statistically significant. Results Compared with group C, H / R group, liver cell culture supernatant ALT, AST, MDA in liver cells was significantly higher (P lt; 0.01), while the liver cells SOD, GSH decreased significantly (P lt; 0.01 ). With H / R group, Wy14643 each dose treatment groups, liver cell culture supernatants of ALT, AST activity was significantly reduced, and dose-dependent manner (P lt; 0.05); Wy14643 10umol / L group, the liver cells The activity of SOD and GSH content increased, while the MDA content decreased, but not statistically significant (P gt; 0.05); Wy14643 30umol / L group, the liver cells of SOD, GSH activity was enhanced (P lt; 0.05 ), while the MDA content decreased (P lt; 0.05); Wy14643 100umol / L group, the liver cells of SOD, GSH activity was enhanced (P lt; 0.01), while the MDA content decreased (P lt; 0.01), significant difference. Compared with group C, H / R group cell activity, PPARαmRNA expression was significantly decreased (P lt; 0.01); with H / R group, Wy14643 10umol / L group, Wy14643 30umol / L group with increasing concentration, PPARαmRNA expression and cell activity increased (P lt; 0.05 or P lt; 0.01); Wy14643 100umol / L group significantly increased cell activity, PPAR-αmRNA expression was significantly increased, significant difference (P lt; 0.01). Electron microscopy showed: Compared with group C, H / R group was significantly mitochondria, membrane damage, with swollen feet and vacuolar degeneration, crest destruction, less rough endoplasmic reticulum, lysosomes and ribosomes less. With H / R group, Wy14643 each dose group of mitochondria, rough endoplasmic reticulum, lysosomes and ribosomes changed significantly reduced with increasing dose. Conclusion Wy14643 liver hypoxia / reoxygenation injury in rats, the mechanism may be by upregulating the expression of PPAR-α, enhanced cellular resistance to antioxidant capacity.
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