基础医学院知识库

目的：本文通过维拉帕米、马索罗酚和阿帕替尼等药物对包虫生长发育的研究，探讨钙信号通路和宿主血管生成信号通路与包虫生长发育之间的关系，期待发现新的抗包虫病药物靶标和筛选出新的抗包虫病候选药物。
方法：药物干预原头节后，评价其作用效果并观察原头节的形态变化；建立包虫病小鼠模型，在不同感染阶段和干预治疗前后分别收集小鼠血清并分装冻存，组织脏器和包囊进行称重，固定储存等，并运用 ELISA、IHC-P、AFS、RT-PCR和 SEM-EDX 等方法对其进行检测分析。
结果：1）与空白组比较，对照组小鼠不同感染期血清和肝脏中钙的含量明显降低(P<0.05)，而 VEGF 及 CD34（微血管密度）的表达量却明显升高(P<0.05)，肝脏胶原纤维的含量也明显升高。2）与对照组比较，维拉帕米干预组小鼠血清及肝、肺等组织中钙的含量明显升高(P<0.05)；维拉帕米可明显下调泡型包虫病小鼠血清和肝脏中 CAM 和CAMK2 的表达(P<0.05)；维拉帕米体内外可分别抑制泡球蚴和细粒棘球绦虫原头节的生长发育（抑囊率和死亡率均明显升P<0.05）。3）马索罗酚体内外可分别抑制泡球蚴/棘球蚴和细粒棘球绦虫原头节/成虫的生长发育（抑囊率和死亡率均明显升高，P<0.05）；对照组泡球蚴表面布满血管，马索罗酚组泡球蚴表面血管明显减少；与对照组比较，马索罗酚组小鼠血清中 VEGF 和 VEGFR 的含量、肝脏中 VEGF 和 VEGFR 的含量及 VEGF-mRNA 的表达量均明显降低(P<0.05)；马索罗酚可明显抑制泡型包虫诱发宿主病理性肝纤维化。4）阿帕替尼体外可明显抑制多房棘球绦虫原头节的生长发育（死亡率明显升高，P<0.05）。
结论：包虫的生长发育与钙离子密切相关，钙信号通路将有望成为抗包虫病的重要靶标；包虫的生长发育可诱导宿主大量的血管生成，其可被马索罗酚抑制，血管生成信号通路将有望成为抗包虫病的重要靶标；维拉帕米、马索罗酚和阿帕替尼均可明显抑制包虫的生长发育，有望成为抗包虫病的候选药物。

Objective: To discover some new targets of drug against echinococcosis or screen out new candidate drugs against echinococcosis on researching pharmacodynamics of verapamil, masoprocol and apatinib against echinococcosis in the hope of probing the relation between growth and development of hydatid and Calcium/angiogenesis signal pathway.
Methods: To evaluate efficacy of the drugs and observe the morphological changes protoscoleces (PSCs) of Echinococcus granulosus (E.g) and E. multilocularis (E.m) cultured with various drugs. Serum and organs in different period were collected from the eastablished BALB/C mice model of Echinococcosis by inoculated with E.g/E.m-PSCs after treatment with diiferent drugs.Weight of the cyst/organs were measured and the samples including serum, tissues were analysed by using ELISA,IHC-P, AFS, RT-PCR and SEM-EDX.
Results: 1) In contrast to blank group, the level of calcium of mice’s serum and liver decreased significantly, the expression of VEGF and CD34 (an indicator of microvessel density) and hepatic collagen fibers increased obviously in control group.2) In contrast to control group, the level of calcium of mice’s serum and organs including liver and lung increased significantly in verapamil (verp) group (P<0.05). Verapamil down-regulated significantly the over-expression of CAM and CAMK2 from mice’s serum and organs (P<0.05). Verapamil inhibited significantly the growth and development of alveococcus in vivo and E.g-PSCs in vitro through increasing the efficacy on alveococcus and the mortality rate of E.g-PSCs (P<0.05).3) Masoprocol inhibited significantly the growth and development of hydatid in vivo and E.g-PSCs/adult in vitro through enhancing the efficacy on hydatid and the mortality rate of E.g-PSCs/adult (P<0.05). Masoprocol reduced abundantly pathological angiogenesis on the surface of E.m-metacestode. Masoprocol decreased obviously the aberrant expression of VEGF/VEGFR in the serum and VEGF/VEGFR/VEGF-mRNA in liver (P<0.05). Masoprocol inhibited significantly pathological hepatic fibrosis of host infected with E.m-PSCs.4) Apatinib significantly inhibited the growth and development of E.m-PSCs in vitro through increasing the mortality rate of E.m-PSCs (P<0.05).
Conclusion: Calcium signaling pathway tightly linked with growth and development of hydatid will be an important drug-target against echinococcosis. Overactive angiogenesis signal pathway triggered by growth and development of hydatid can be inhibited by masoprocol and will be a significant target against echinococcosis. The drugs including verapamil, masoprocol and apatinib, which can inhibit growth and development of hydatid, will be candidate agents against echinococcosis.