缺氧对肝泡状棘球蚴原头节血管内皮生长因子和CD34表达的影响研究

摘要

Objective To investigate the expression level of vascular endothelial growth factor(VEGF) and CD34 in the hepatic alveolar echinococcosis protoscolex under the condition of vitro anoxia.Methods From April to October in 2016,the 30 cases gerbils of abdominal infection with hepatic alveolar echinococcosis for 18 weeks were selected for experiment,and were sacrificed by cervical dislocation.The smooth clusters of hepatic alveolar echinococcosis tissues isolated in abdominal cavity or tissues adhere to the abdominal wall,mesentery,liver surface were selected.The hepatic alveolar echinococcosis protoscolex was cultured to divide into control group and experiment group(RPMI 1640 medium containing 100 μmol/L cobalt chloride).The hepatic alveolar echinococcosis protoscolex activity was observed by inverted microscope at the beginning,and the 6th,12th,18th,24th hour of the culture process.At the 12th and 24th hour of the culture process,enzyme-linked immunosorbent assay(ELISA) was used to detect the expression level of hypoxia-inducible factor 1 alpha(HIF-1α),VEGF and CD34 in hepatic alveolar echinococcosis protoscolex.Western blotting method was applied to detect the expression of VEGF and CD34.Results The echinococcosis protoscolex activity at the 0th,6th,12th,18th,24th hour of the culture process was not significantly different between control group and experiment group(P>0.05).Compared with the control group,the expression level of HIF-1α,VEGF,CD34 were significantly increased at the 12th and 24th hour of the culture process in the experiment group(P<0.05).Conclusion Hypoxia can increase the expression level of VEGF and CD34 in echinococcosis protoscolex,and this may be one of the important mechanisms of invasive growth presented by hepatic alveolar echinococcosis.%目的 探讨体外缺氧条件下肝泡状棘球蚴原头节中血管内皮生长因子(VEGF)、CD34表达水平的变化.方法 2016年4-10月,选取实验用腹腔感染肝泡状棘球蚴18周的长爪沙鼠30只,采用颈椎脱臼法处死,选取光滑、成簇的游离于腹腔或附着在腹壁、肠系膜、肝表面的肝泡状棘球蚴组织.培养肝泡状棘球蚴原头节,分为两组,对照组和实验组(给予含100 μmol/L二氯化钴的RPMI 1640培养基),培养第0、6、12、18、24 h倒置显微镜下观察肝泡状棘球蚴原头节活力.培养12、24 h采用酶联免疫吸附试验(ELISA)法检测肝泡状棘球蚴原头节中缺氧诱导因子1α(HIF-1α)、VEGF、CD34表达水平,Western blotting法检测肝泡状棘球蚴原头节中VEGF、CD34的表达.结果 培养0、6、12、18、24 h,对照组与实验组肝泡状棘球蚴原头节活力比较,差异均无统计学意义(P>0.05).培养12、24 h,实验组肝泡状棘球蚴原头节HIF-1α、VEGF、CD34表达水平较对照组升高(P<0.05).结论 缺氧可以使肝泡状棘球蚴原头节中VEGF和CD34表达水平升高,这可能是肝泡状棘球蚴病呈浸润性生长的重要机制之一.