Gene therapy by hepatocyte growth factor results in regression of experimental liver fibrosis

Aim of investigation. Studying of efficacy of genetic therapy of liver fibrosis at mice by human hepatocyte growth factor (HGF) and evaluation of potentials of hydroporation method for delivery of genetic complexes to the liver.

Material and methods. During experiment male mice of Balbc line received intraperitoneal injections of 30% perchloromethane (CCl4) diluted in oil in a dose of 1 ml/kg once per week. Animals (n=4–6) dropped from the experiment at the 1, 2, 4 and 6-th week from its onset. After obtaining liver tissue samples, histological study was carried out, and by real time polymerase chain reaction (RT-PCR) contents of mPNA of liver fibrosis-associated genes were analyzed: transforming growth factor β1 (TGF-β1), collagen 1α1 (Coll1α1), matrix metalloproteinase-13 (ММР-13), tissue inhibitor of matrix metalloproteinase (TIMP-1) and smooth-muscle actin α (α-SMA). A blood was extracted for assessment of liver enzymes activity (AST and ALT). Human HGF was injected into liver tissue by hydroporation method. Contents of human HGF in mice liver tissue was detected by PCR. For evaluation of stage of liver fibrosis Knodell index and morphometrical analysis was used.

Results. According to Knodell index development of CCl4-induced liver fibrosis at the 4-th week of experiment was found. Liver cirrhosis developed at the 6-th week of experiment. These data correlated well to the RT-PCR results and evaluation of liver enzymes activity. Morphometrical analysis demonstrated, that after HGF therapy in the main group of animals the degree of fibrosis (%) was lower in comparison to that in the control group. Efficacy of transfection of complementary HGF DNA at application of hydroporation method was higher, than at direct method of injection into parenchyma of the organ.

Conclusions. Obtained results allow to consider gene therapy by hepatocyte growth factor in composition of non-viral vectors as one of perspective methods of treatment of chronic liver diseases.

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