The In Vivo And In Vitro Experimental Study On Effect Of VEGF And CD34 In The Growth Of Hepatic Echinococcus Multilocularis

Objective: The aim of this study is to detect the expressions of CD34 and VEGF in In vivo and in vitro expertmental of echinococcus multilocularisMethods: In our first expertmental,we established a co-cultrue systerm used alveolar hydatid protoscolex and human hepatoma Hep G2 cell.After 7 days co-culture, the cultured alveolar hydatid protoscolex divided into three groups, deferoxamine hypoxia-inducible group, DMSO vehicle group, the conventional training group. After next 24 hours, we counted the nortality of the protoscolex by Eosin staining.Then,using the Elisa method to detect the expression of HIF1-α,VEGF and CD34. In our second expertmental, Sixty health gerbils were randomly equally divided into two groups, an experimental group and a sham operation group, each gerbil was given liver vaccination by opening their abdominal. Each gerbil in the experimental group was injected with approximately 400 echinococcus protoscoleces(0.1 m L), and each gerbil in the sham operation group received a corresponding volume of physiological saline. Six gerbils were sacrificed on day 20, 40, 60, 80, and 100. The hepatic alveolar hydatid tissue(AE) and its surrounding liver tissue(HSAE) were collected from the experimental group and the normal liver tissue(NL) was collected from the sham operation group, and the expressions of microvessel density(MVD)-CD34 and vascular endothelial growth factor(VEGF) were detected by immunohistochemistry staining(En Vision method).Results: After 24 hours’ s culture,the mortality rate of the normal group, DMSO vehicle group and deferoxamine hypoxia-inducible group presented as follows:(79.90±1.31)%,(78.58±2.52)% and(78.45±1.65)%,there was no significant difference between the groups(P>0.05)。The expression of HIF1-αpresented here:the normal group was:(852.00+124.41)pg/m L,DMSO vehicle group was:(806.20±118.51)pg/m L and deferoxamine hypoxia-inducible group was:(1168.20±93.59)pg/m L.The expression of VEGF presented here:the normal group was:(158.28±29.24)pg/m L,DMSO vehicle group was:(157.49±26.88)pg/m L,deferoxamine hypoxia-inducible group was(222.36±18.87)pg/m L.The expression of CD34 presented here: the normal group was(605.40±49.95)pg/m L,DMSO vehicle group was:(592.20 ±51.91)pg/m L and deferoxamine hypoxia-inducible group was:(873.60±61.15)pg/m L.Compared with the normal group and DMSO group,The expression of HIF1-α、VEGF、CD34 were significance differences(both P<0.05);Compared with DMSO vehicle groupand deferoxamine hypoxia-inducible group,there were no statistically significant of the expression of HIF1-α、VEGF、CD34(both P>0.05)。E.multilocularis hydatid tissues were observed over the liver and in the abdominal cavity in the experimental group each gerbil by general observation. The expressions of CD34 and VEGF were observed in the AE at each timepoint after infection and located in the cytoplasmic of endothelial cells. The number of MVD-CD34 of AE at each timepoint in the AE was(9.83±3.87) /HP,(25.33±6.71)/HP,(34.50±5.50)/HP,(37.67±5.71)/HP and(44.67±4.93)/HP, respectively, which were significantly higher than those in the HSAE 〔0/HP,(1.17±0.98)/HP,(3.50±1.38)/HP,(5.83±2.71)/HP, and(8.83±2.48)/HP, repectively〕 and NL(all were 0), P<0.05. The points of VEGF at each timepoint in the AE was 2.95±0.46, 3.90±0.68, 4.27±1.05, 5.33±0.95, and 4.50±0.81 respectively, which were significantly higher than those in the HSAE(1.07±0.63, 1.38±0.75, 1.55±0.83, 1.67±0.47, 2.10±0.55, repectively〕 and NL(1.02±0.83, 1.12±0.63, 1.26±0.26, 1.20±0.74, 1.21±0.28), P<0.05.Conlusions: 1. We were successfully established in vitro hypoxia culture model of protoscolex and there were no significance of the protoscolex’s mortality rate after 24 hours hypoxia clutured;2. The expression of HIF1-α、VEGF、CD34 were upregulated after 24 hours hypoxia coulture; 3 The overexpression of VEGF and CD34 may contribute to gerbils alveolar hydatid tissue angiogenesis; 4. Angiogenesis may be one role of the invasion and metastasis of alveolar hydatid。...