The Effects Of Exogenous Human VEGF165Gene On Liver Regeneration In Hepatocirrhosis After Hemihepatectomy In Rat

Objective: To study the effects of exogenous hVEGF165on liver regeneration andthe recover of liver function in hepatocirrhosis after partial hepatectomy in rats. Methods:The hVEGF165gene was cloned to lentiviral expression vector by recombinant DNAtechnology, the positive clones were screened and lentiviral packaged systems(ViraPowerTMPackaging Mix) were co-transfected to package virus in293T cells bylipofection with target gene plasmid. Using the Real time PCR technique to test the titer ofpLenti6/V5-D-TOPO-hVEGF165. The BLR3A rat liver cells were transfected bypLenti6/V5-D-TOPO-hVEGF165and the expression of hVEGF165gene was detected byRT-PCR and Western blot technique; Building cirrhotic model in150rats by traditionalCCl4induced method, model is validated with liver biopsies. The126cirrhotic rats afterhemihepatectomy was randomly divided into three groups, group A (VEGF165group):portal vein was injected with the lentivirus vector containing hVEGF165; Group B (emptyvirus group): portal vein was injected with empty lentivirus vector; Group C (controlgroup): portal vein was injected with PBS buffer. Expression of hVEGF165mRNA andprotein in liver tissue, liver regeneration rate, the positive rate of PCNA, ALT and ASTcontent in the blood of portal vein were detected before and12h,24h,72h,7d,14d,28dafter operation. Results: The lentivirus vector containing hVEGF165was successfullyconstrected. Virus titer could reach as high as1.18×10~7v.p./mL. The positive rate of greenfluorescent protein was more than80%in the BLR3A rat liver cells after transfecting withpLenti6/V5-D-TOPO-hVEGF165in72h, RT-PCR and Western blot results showed thatthe hVEGF165gene expression of the transfected group was positive, but the negativecompared group and blank compared group were negative. The shaping rate of cirrhoticmodel in rats was96.18%. The liver regeneration rate (34.98±6.22%,62.19±1.81%,83.54±4.50%and92.23±4.41%) during72h to28d after operation and the positive rate of PCNA (18.05±0.92%,42.89±5.52%,35.56±3.86%,26.37±1.35%and19.48±2.70%)during24h to28d after operation of VEGF165group were obviously higher than those inthe control group (P<0.01); the serum concentration of ALT (258.14±12.94U/L,215.67±22.12U/L,175.92±8.78U/L and133.26±13.92U/L) and AST (553.34±14.10U/L,481.37±14.74U/L,425.95±5.04U/L and388.29±15.99U/L) during72h to28d afteroperation of VEGF165group were evidently lower than those in the control group(P<0.01). Conclusion: The exogenous hVEGF165has significantly positive effect onliver regeneration and the recover of liver function after partial hepatectomy in rats withhepatocirrhosis.