Effect Of COX-2ShRNA On The TGF-β1/SMAD Signal Transduction Pathway Of Hepatic Fibrosis

ObjectivesTo explore the influence of COX-2ShRNA on the TGF-β1/SMADsignaling pathway of rat’s hepatic fibrosis, in order to provide newideas for clinical treatment of liver fibrosis.Methods50specific pathogen-free male and healthy SD males rats wererandomly divided into five group:normal controls group (Agroup), model group (B group), COX-2ShRNA group (C group),empty vector group (D group), nimesulide group (E group). Rat’smodels of hepatic fibrosis in group B, C, D, and E were establishedthrough pouring high fat diet and40%CCL4peanut oil,2kgml/5d, intostomach. Starting from the first week of modeling, for each rat ingroup C, transfect ion in vivo was conducted through tail veininjection COX-2ShRNA recombinant adenovirus vector（titer1×1010PFU/ml）100ul per day; rats in group D were injected of equal nullrecombinant vectors not containing COX-2ShRNA gene; in group E,2ml/kg d nimesulide was poured into stomach. Rats in the normalgroup and model group were given of an equal volume of saline; at theend of the12thweek,12h after giving medicine for the last time, the rats were killed. Observe the inflammation of hepatic issue and thedegree of fibrosis of rats in each group through HE and MASSONdyening. Observe rat’s hepatic expressions of COX-2、TGF-β1andSMAD1-5’s mRNA expression and protein expression throughRT-PCR and immunized test, Use the SPSS17.0statistics soft toanalysis the experimental data.Results1.Results of liver wet weight and hepatic indices: Compared withrats in group A, rats in group C had increased hepatic wet weight andindice（sP＜0.05）; Compared with rats in group B、D, rats in group C haddeclined hepatic wet weight and indices（P＜0.05）.2. Serum biochemical test results: Compared with rats in group A,rats in group C had higher level of serum AST、ALT、TC、TG（P＜0.05）;Compared with rats in group B and D, rats in group C had lower level ofserum AST、ALT、TC、TG（P＜0.05）.3. Results of HE、MASSON staining: Compared with rats in groupA, changing degree of liver fibrosis for rats in group C is obviously moreserious（P＜0.05）; Compared with rats in group B and D, changingdegree of liver fibrosis for rats in group C obviously reduces.（P＜0.05）.4. Quantitative inspection and immunohistochemical method resultsof COX-2、TGF-β1and SMAD1-5RT-PCR: In group C, the COX-2、TGF-β1and SMAD1-5mRNA expression in rat’s hepatic issue had a obvious higher level of expression than those in group A（P＜0.05）;COX-2、TGF-β1and SMAD1-5’s mRNA in rat’s hepatic issue in group Chad a lower level of expression compared with those in B and D（P＜0.05）.5.Correlation analysis between COX-2mRNA expression andTGF-β1、SMAD1-5’s protein expression:There was a positive correlationbetween COX-2’s mRNA expression and TGF-β1、SMAD1-5’s mRNAexpression. There was a positive correlation between COX-2’s proteinexpression and TGF-β1、SMAD1-5’s protein expression.Conclusion1.COX-2ShRNA can effectively lower the expression of COX-2in rat’s hepatic issue.2.COX-2ShRNA can produce anti-hepatic fibrosis effect throughcontrolling the expression of gene such as COX-2、 TGF-β1、SMAD1-5and so on in rat’s hepatic issue.