RESUMO
Objective To study the content of IL-8 in the peripheral blood and the expression of its mRNA in liver biopsy tissue of patients with hepatocirrhosis.Methods The patients with typical hepatocirrhosis resulted from HBV infection were selected.The IL-8 concentrations in peripheral blood of the Datients were evaluated with ELISA.The content of IL-8 mRNA in liver biopsy was measured by real-time PCR.The level of IL-8 mRNA was surrogated by lg cDNA/lg GAPDH.Reslllts The levels of IL-8 in the serum and its mRNA in hepatic tissue of patients with hepatoeirrhosis were(43 1.39±97.39)μg/L and 1.3647±0.2203,respectively,which were significantly higher than those of control group.MoreoCer,the levels of IL-8 in the serum and IL-8 mRNA in hepatic tissue of patients with positive HBV-DNA were (502.43-4-102.24)μg/L and 1.51424±0.2245,respectively,and both ofwhich were higher than those of patients without HBV-DNA replication(P<0.0l,P<0.05).And the levels of IL-8 in the selqlm and its mRNA in hepatic tissue of patients with higher ALT.AST also increased significantly.compared with those in control group(P<0.01).Conclusion The levels of IL-8 in the peripheral blood and the IL-8 mRNA in liver biopsy tissue of patients with hepatocirrhosis increase.which are related to the degree of HBV-DNA replication and the damage of hepatocytes.The IL-8 plays a key role in local inflammatory infihration of liver and the progression of post-hepatitis cirrhosis.
RESUMO
Objective:To establish a method which could quantify human IL-8 mRNA in peripheral blood monocytes.Methods:Two probe were designed aiming at amplicon of RT-PCR,of which one being capture probe adorned 5,end by ammonia and one being monitor probe adorned 3,end by bintony.So as,the capture probe could couple with the N-oxysuccinmide esters(NOS) group on the DNA-binding wells through covalent attachment.In this way,oligonucleotides were immobilized on the plate surface and arise straightly to capture target amplicon.Total RNA was extracted from peripheral blood mononuclear cell and IL-8 mRNA was amplified by using RT-PCR.Heating denatured products and denatured probe,were then added to wells and hybridization occurs between capture probe,target amplicon and detection probe.After the addition of Avidiu-peroxidase developing system,the optical density were read at 450 nm.Results:Sensitivity,detecting PCR product of 16 PCR cycling,of 5?10~3 PBMCs and of a dilute 1∶256 were its prior.Meanwhile,this method could get a specify result and a precise of 5.2%.Conclusion:Resulting in simplicity,sensitivity and specificity results,this method may be a good choice for quantify of PCR products.