كلونينگ، بررسي توالي و بيان كموكين آلفا SDF-1 در سلول هاي كبدي موش صحرايي

Cloning, Sequencing and Evaluation of Expression of SDF-1 a in Rat Hepatocytes

Background and purpose: High constitutive levels of SDF-1a have been observed in the non-inflamed biliary epithelium of the liver. SDF-1a is also produced by ductal plate cells (biliary epithelial cells progenitors) and it is involved in maturation and homing of B cells in fetal liver. Giving the above brief introductory remarks, this chemokine was chosen to analysis in isolated and cultured hepatocytes. Materials and Methods: We employed gene cloning methods to clone and northern analysis to show the expression of the gene in hepatocytes. In the first stage of this work, we have sequenced cloned fragment of SDF-1 in cultured rat hepatocytes and compared with database to prove that the studied and cloned gene is SDF-I. Results: we showed that the fragment, which we have used as probe for our northern blotting analysis is SDF-1 by sequencing methods. Following using specific produced probe for SDF-1, in this study we showed that SDF-1 is expressed by hepatocytes after isolation and early culture of hepatocytes. Conclusion: Isolation of hepatocytes by enzymatic methods causes liver injury and therefore hepatocytes may stimulate to response to the injury. The expression of this chemokine may involve in stress response of hepatocytes to isolation.