The Effect Of VEGF ShRNA Plasmid And CD34 ShRNA Plasmid On Angiogenesis In The Infiltrating Growth Of The Gerbil Hepeatic Alveolar Hydatid Was Compared

Objective Regulated down VEGF and MVD-CD34 expression in the liver alveolar echinococcosis of the gerbil by using RNA interference technique,and observe the effects of the observation on angiogenesis in the growth of hepatic vesiculae,and in order to provid new ideas which the control of the proliferation of alveolar echinococcosis in vivo.Methods According to the gerbil VEGF and CD34 length cDNA sequence,we synthesis specificity silent corresponding mRNA six siRNA oligonucleotide fragments respectively,and construct recombinant plasmid,and screen plasmid which is most efficient interference by RT-PCR,and with liposomes in proportion to inject the tail vein in gerbil after amplificating and extracting plasmid.50 healthy gerbils are inocolated of alveolar echinococcosis in percutaneous puncture right hepatic and dividedinto 5 groups after 30 days randomly,they are injecting VEGF plasmid group(VEGFgroup),injecting CD34 plasmid group(CD34 group),injecting VEGF-CD34plasmid group(combination group),injecting liposome group(liposome group)and treatment group,each group of 10,injecting 200?g to the vein tail of VEGFgroup?CD34 group and liposome group,every 5 days,and Injecting 400?gcombination group group every 5 days,treatment group isn't inject,there are 10 normol gerbils as blank group;at 90~thh day,taking the whole blood in aorta and weighing abdominal alveolar echinococcosis include all clusters bubble ball larva on the liver tissue,and calculat suppressing rate,and formaldehyde fixed respectively each liver alveolar echinococcosis tissue?border area tissue and normal liver tissue,and observe pathology differences by eosin stain;Observe and evalute the expression of VEGF and CD34 in each tissue by mmunohistochemical method;determine the serum VEGF and CD34 by Enzyme linked immunosorbent assay(Elisa).Results RT-PCR showed that VEGF shRNA3 and CD34 shRNA1 plasmid had the strongest inhibitory effect on mRNA in the liver of the gerbil;three capsule wet weight of plasmid group were lower than the treatment group:joint group<VEGF group<CD34 VEGF<treatment group,the comparative differences of two of them are statistically significant(P<0.05),the capsule wet weight of liposome group had no difference with the treatment group(P>0.05);compared with the treatment group,there was no significant change in the structure of cuticle,germinal laye and granuloma in hematoxylin and eosin stain;VEGF score of normal liver tissue of every groups is roughly same,VEGF score of liposome group and the control group with same tissue have no difference of statistical significance in immunohistochemical,(P>0.05),VEGF expression of the plasmid group in liver alveolar echinococcosis tissue and border area tissue are lower than the control group,the difference was statistically significant(P<0.05),the comparison among three plasmid groups in liver alveolar echinococcosis tissue and border area tissue,VEGF score from low to high followed by:combination group<VEGF group<CD34 group,it has marked difference(P<0.05);There is no CD34 expression in normal liver tissue,every issue of liposome group and treatment group have same expression level of CD34 expression,but CD34 expression levels of liver alveolar echinococcosis tissue and border area tissue were significantly lower than treatment group,the difference was statistically significant(P<0.05),the comparison among three plasmid groups in liver alveolar echinococcosis tissue and border area tissue,CD34 expression from low to high is:combination group<VEGF group<CD34 group;ELISA result showed that VEGF and CD34 expression of each group gerbils in serum have different level,VEGF and CD34 of liposome group and combination group have no statistically significant difference,compared with treatment group,serum VEGF and CD34 of combination groupwere lower(P<0.05),comparing differences among plasmid group has significance(P<0.05),comparison of VEGF and CD34 is:combination group<VEGF group<CD34 group.Conclusion 1.VEGF shRNA,CD34 shRNA plasmids can reduce VEGF and CD34 expression of in the liver alveolar echinococcosis of the gerbil,inhibit the growth of its angiogenesis of invasion growth effectively,delay the growth speed of the liver alveoar echinococcosis;2.VEGF shRNA plasmids effect on the liver alveolar echinococcosis is higher than that of CD34 plasmid.