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Objective: To detect liver tissue heme oxygenase -1 (HemeOxygenase, HO-1) mRNA and HO-1 protein expression and apoptosis after ischemia-reperfusion explore the liver tissue response to ischemia-reperfusion HO-1 relationship between the protein and hepatocyte apoptosis. , Inhibit apoptosis, reduce hepatic ischemia-reperfusion injury (HepaticIschemic-Reperfusion Injury, HIRI) provide a theoretical basis for the selection of effective pretreatment method. Methods: 1, specimen collection: liver incision, respectively hilar occlusion, Pringle take the cutting edge of the next 2cm liver tissue 10g open at the same time, before the end of surgery, the abdomen was closed. Divided into two, one with 4% paraformaldehyde, paraffin specimens. Save another placed at -70 ℃ for extraction of total RNA. Another liver cut before the start of surgery, after the first four or seven days to take patients with peripheral venous blood 5 ml of. 2, reverse transcriptase polymerase chain reaction (RT-PCR) assay HO-1 mRNA expression. Deoxynucleotidyl transferase mediated nick end labeling (Terminaldeoynucleotidletransferase the mediated d-UTP nick end labeling, TUNEL) assay hepatocyte apoptosis in liver tissue. 4, immunohistochemistry method (SP) Determination of the expression of HO-1 protein in the liver tissue. 5, measured with an automatic biochemical analysis of plasma alanine aminotransferase (Alanine aminotransferase, ALT), aspartate amino transfer enzyme (Aspartate inotransferase, AST) and total serum bilirubin (Total Bilirubin, TB). Results: 1, the liver tissue of HO-1mRNA the expression, Pringle liver doors open 0.2313 ± 0.0575, 0.8168 ± 0.1972, the abdomen was closed before as 1.2447 ± .4403. The differences between the three groups was statistically significant (p <0.05). Pringle former HO-1 mRNA expression reduce than liver doors open, the abdomen was closed before the increase of HO-1 mRNA expression than liver doors open. 2, hepatocyte apoptosis, apoptotic index before Pringle (Apoptotic Index, AI) was 9.59 ± 0.959 ‰, liver doors open at 5.80 ± 0.939 ‰, the abdomen was closed prior to 4.69 ± 1.738 ‰. Hepatocyte apoptosis index in the difference between the three groups was statistically significant (p <0.05). Hepatocyte apoptosis index higher than liver Pringle door open and the abdomen was closed; hepatic portal open hepatocyte apoptosis index higher than before the abdomen was closed. 3, HO-1 protein expression with positive area and integrated optical density (Integrated OpticalDensity, IOD). Pringle 63158.70 ± 7444.44, liver doors open to 117,552.10 ± 18559.50 and 146,580.30 ± 47256.92 before the abdomen was closed, and the change in the three groups, the difference was statistically significant (p <0.05). HO-1 expression in liver tissue IOD Pringle 9551.58 ± 2508.77, liver doors open for 27665.29 ± 4252.36 abdomen was closed before 33790.16 ± 6574.92, changes in the difference between the three groups was statistically significant (p < 0.05). Positive area and IOD HO-1 expression in the liver tissue increased significantly before the abdomen was closed than liver doors open, to than before Pringle increase in the hepatic portal open. 4, ① ALT among the three groups the difference was statistically significant (p <0.05). ALT in the first postoperative day, the fourth day, the seventh day are increased (p <0.05), and after the first day of ALT reached a peak, and then gradually decline. ② AST, the difference between groups was statistically significant (p <0.05), AST reached a peak in the first postoperative day, followed by a gradual decline. After the seventh day and the difference was not statistically significant (p> 0.05) compared to before the surgery. The ③ TB group asked the difference was statistically significant (p <0.05), and gradually increased during the first postoperative day, postoperative fourth day reached a peak, and postoperative seventh day after the first day The difference was not statistically significant (p> 0.05). 5, correlation analysis: ① hilar blocking before liver tissue HO-1 mRNA expression in liver tissue HO-1 protein correlation coefficient γ = 0.986, p = 0.000 (p <0.05); were highly positively correlated. ② Pringle before liver tissue HO-1 mRNA expression and hepatocyte apoptosis correlation coefficient γ = -0.961, p = 0.000 (p <0.05); showed a high negative correlation. ③ liver doors open, HO-1 mRNA expression in liver tissue and liver tissue HO-1 protein correlation coefficient γ = 0.985, p = 0.000 (p <0.05); were highly positively correlated. ④ liver doors open, the correlation coefficient of the HO-1 mRNA expression in liver tissue and liver cell apoptosis γ = -0.983, p = 0.000 (p <0.05); showed a high negative correlation. ⑤ abdomen was closed before the liver tissue expression of HO-1mRNA liver tissue HO-1 protein correlation coefficient γ = 0.996, p = 0.000 (p <0.05); were highly positively correlated. ⑥ abdomen was closed HO-1 mRNA expression in liver tissue and liver cell apoptosis correlation coefficient γ = -0.962, p = 0.000 (p <0.05); showed a high negative correlation. Conclusion: 1, only a small amount of HO-1 mRNA in normal liver tissue and HO-1 protein expression after liver ischemia-reperfusion injury in liver tissue HO-1 mRNA and HO-1 protein were significantly higher expression, and an upward trend, suggesting that HO-1 has a certain relationship with the occurrence and development of HIRI. 2, Pringle caused by ischemia and reperfusion injury after liver tissue HO-1 mRNA and HO-1 protein expression increased hepatocyte apoptosis index decreased. Description of HO-1 mRNA in the liver tissue and increased expression of HO-1 protein may have a certain inhibition of liver cell apoptosis.
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