靶向超声造影在体定量评价大鼠移植后卵巢新生血管

Targeted contrast enhanced ultrasound for in vivo quantitative evaluation on angiogenesis in transplanted ovarian of rats

目的观察以抗苗勒管(AMH)激素靶向纳米泡(AMH-NB)为超声造影剂在体定量评价大鼠自体卵巢移植后新生血管密度的价值。方法制备AMH-NB并检测其基本物理特性。建立大鼠自体卵巢移植模型,于术后第7天行靶向(AMH-NB)、非靶向(N-NB)及声诺维(SonoVue)超声造影,获得峰值强度(PI)和达峰时间(TTP)。以免疫组织化学法检测微血管密度(MVD),并对PI、TTP与MVD进行相关性分析。结果所制备的AMH-NB粒径(622.67±33.65)nm,分布均匀,浓度(2.90±0.26)×10^8/ml。AMH-NB超声造影显示卵巢PI为(7.93±0.65)dB,TTP为(42.53±1.74)s;N-NB造影PI为(6.14±0.44)dB,TTP为(54.35±1.73)s;声诺维造影PI为(4.15±0.83)dB,TTP为(28.71±1.18)s(P均<0.05)。免疫组织化学分析显示移植后卵巢微血管密度为(61.20±6.84)/HP,组织学分析AMH-NB可穿过血管内皮细胞间隙进入组织间隙并与AMH结合。AMH-NB造影所示PI、TTP与MVD呈高度相关(r=0.84、-0.84,P均<0.05)。结论利用AMH-NB进行超声造影可实现定性、定量评价大鼠移植后卵巢新生血管。

Objective To observe the value of targeted contrast enhanced ultrasound(CEUS)using anti-Mullerian canal hormone(AMH)targeted nanobubbles(AMH-NB)for in vivo quantitative evaluation on ovarian neovascular density after ovarian auto-transplantation in SD rats.Methods The nanobubbles carrying anti-AMH antibody were prepared,and their basic physical properties were examined.Then ovarian auto-transplantation rat models were established.The targeted(AMH-NB),non-targeted(N-NB)contrast agents and SonoVue were administered on the 7 th day after transplantation to obtain peak intensity(PI)and time to peak(TTP).The microvascular density(MVD)was measured using immunohistochemistry,and the correlation of PI,TTP and MVD were analyzed.Results The particle size of AMH-NB uniformly distributed,ranged(622.67±33.65)nm,and the concentration of AMH-NB was(2.90±0.26)×10^8/ml.PI of ovarian angiography with AMH-NB was(7.93±0.65)dB and TTP was(42.53±1.74)s,with N-NB was(6.14±0.44)dB and(54.35±1.73)s,with Sonovue was(4.15±0.83)dB and(28.71±1.18)s,respectively(all P<0.05).Immunohistochemical analysis showed that the microvascular density was(61.20±6.84)/HP,histological analysis indicated that AMH-NB were able to penetrate blood vessels to the interstitial space and combine with AMH.PI,TTP of AMH-NB were highly both correlated with MVD(r=0.84,r=-0.84,both P<0.05).Conclusion AMH-NB can be used to qualitatively and quantitatively evaluate the angiogenesis in transplanted ovarian of rats in vivo with CEUS.