The Research Of The Alterations Of 11Beta-HSD1 Gene And Protein Expression In Rat Liver With Acute Liver Injury And Their Correlation With Serum Transaminases

OBJECTIVEGlucocorticoids are widely used in clinical, including hydrocortisone, prednisone and methylprednisolone etc. Among them, cortisone and prednisone are originally non-active in vivo, which should be catalyzed to hydrocortisone and prednisolone by 11β-HSD1 at liver. The results of the researches about "whether the activation of cortisone and prednisone are affected in the patients with liver injury", which were advanced in 1970s and 1980s, are not unified, and the conclusions are controversial. Around 1990, the gene of 11β-HSD1 was separated and the protein was purified, but the relative expression researches were not performed in damaged liver. So, in order to find more believable laboratory evidence for the rational administration of glucocorticoids in the patients who with liver injury, we set up experiments for studying the alteration of 11β-HSD1 gene expression and protein content in rats which possess acute hepatic injury and also their correlation with serum transaminases.METHODS1. Model of CCl4 induced acute hepatic injury 36 male SD rats were put into different groups of A、B、C、D、E、 F randomLy in weight, with 6 rats in every group. The groups of A、B、C、D、E、 F, were given respectively intraperitoneal injection according to lmL/kg weight corn oil solution of CCl4 for different volume ratios (0、0.0625、0.125、0.25、0.5、1.0).7 days before and 24h after injection of CCl4 solution,2 mL blood of rats was taken out for detecting of ALT、 AST、albumin, and the livers were taken out for degree measurement of liver pathological damage by HE.2. Gene expressions of 11β-HSD1 in the livers of ratsDetect the relative expressions for 11β-HSD1 toward GAPDH by RT-qPCR.3. Protein content of 11β-HSD1 in the rats liverDetect the expression positions of 11β-HSD1 by immunohistochemical method, and the relative protein content of 11β-HSD1 toward β-tubulin by western blot.4. Statistical methodsPaired t test was progressed in rats body weight、ALT、AST、albumin before and after injection of CCl4, ANOVA was progressed in liver weight、ALT、AST、albumin after injection of CCl4, the gene expression of 11β-HSD1 and the protein content of 11β-HSD1 by groups, and correlation and regression analysis were progressed in the gene expression of 11β-HSD1 and the protein content of 11β-HSD1 with ALT、 AST、albumin after injection of CCl4 by rats by SPSS 17.0.RESULTS1. The model of acute hepatic injury was successfully induced. The serum ALT、AST and albumin of rats altered significantly before and after injection of CCl4 solution (p=0.003,p=0.000,p=0.000); The serum ALT、AST of rats after injection of CCl4 solution are different in groups (p=0.000,p=0.000), but the serum albumin is not (p=0.075).Rat liver biopsy shows that cells around the central vein were damaged, and the cytolysis number and area increases along with the increasing of CCl4 dosage.2. The gene expression of 11β-HSD1 did not alter significantly in groups(p=0.705), but altered in rats. The gene expression of 11β-HSD1 correlated with serum ALT、AST of rats after injection of CCl4 solution (r=-0.36,p=0.02、r=-0.324,p=0.035), but didn’t with serum albumin (p=0.998)3. The protein content of 11 P-HSD 1 in the rats liver changed significantly between groups by immunohistochemistry (p=0.013) and by western blot (p=0.000).Along with the liver damaged increasingly, several cells around the central vein apoptosised, while the protein content of 11β-HSD 1 aggravated slightly, and then many cell apoptosised, while the protein content of 11β-HSD 1 got to the maximum,finally to many cells apoptosised, while the content of 11β-HSD1 declinded. The protein content of 11β-HSD1 in the rats liver didn’t correlate with serum ALT、AST、albumin of rats after injection of CCl4 solution (p=0.07, p=0.07,p=0.334).And it reduced after the first increase along with the rise of serum ALT and AST.4.The gene expression of 11β-HSD 1 negatively correlates with the protein content of 11β-HSD1 (r=-0.318, one side p=0.401)CONCLUSIONS1. The expression of 11β-HSD 1 gene altered in rat liver with liver injury, which declines along with the degree of liver damage aggravating, negatively correlating with serum ALT, AST.11β-HSD 1 gene expression decreased to half when ALT reaches 111.0 IU/L or AST to 97.5 IU/L.2. The content of 11β-HSD1 protein altered in rat liver with liver injury, which was firstly increased and then declined along with the degree of liver damage aggravating.11β-HSD1 protein content reach the summit when ALT becomes 102.9 IU/L or AST to 92.1 IU/L3.11β-HSD1 protein expression area changed in rat liver with the damaged liver, which went away from the central vein along with the degree of liver damage increasing.4. The alteration of 11β-HSD1 gene expression negatively correlated with the alteration of protein content when the liver injured;...