Intra-Seminiferous Tubular Injection of Vascular Endothelial Growth Factor C Sustained-Release Ultrafine Particles: A Novel Method for Improving the Regeneration of Spermatogenesis After Chemotherapy.
Intra-Seminiferous Tubular Injection of Vascular Endothelial Growth Factor C Sustained-Release Ultrafine Particles: A Novel Method for Improving the Regeneration of Spermatogenesis After Chemotherapy.
复制标题
生精管内注射血管内皮生长因子C缓释超细颗粒：改善化疗后生精再生的新方法
DOI：
10.1166/jbn.2019.2857
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发表时间：
2019
影响因子：
2.9
通讯作者：
Li Zheng
中科院分区：
文献类型：
--
作者：
Zhao Liangyu;Yao Chencheng;Zhu Zijue;Liu Nachuan;Zhai Jing;Wang Yizhou;Yang Chao;Tian Ruhui;Li Peng;Chen Huixing;Wan Zhong;Zhi Erlei;Huang Yuhua;Zhou Zhi;Li Zheng
白消安和其他用于治疗癌症的化疗药物可能会对精子发生造成暂时性甚至永久性的损害。在精子发生过程中，快速分裂的精原细胞对化疗非常敏感。因此，人们对开发一种可以在化疗后提供刺激和再生精子发生的方法非常感兴趣。在以前的研究中，我们提出了潜在的应用血管内皮生长因子C（VEGFC），因为它在刺激精原细胞增殖的关键作用。然而，使用VEGFC或其他调节因子促进此类患者精子发生恢复的方法非常缺乏，因为这些蛋白质的快速降解和血-睾丸屏障产生的限制。为此，我们将VEGFC负载到聚阳离子壳聚糖壳层中的聚阴离子硫酸葡聚糖中，以制备VEGFC缓释超细颗粒（UFPs，CS-DS-VEGFC）。我们在使用白消安建立的无精子小鼠模型中测试了这种颗粒。对于每只小鼠，将CS-DS-VEGFC注射到一个睾丸的生精小管中，而将未加载的UFP（CS-DS）或单独的VEGFC蛋白注射到相对的睾丸中作为对照。5周后处死所有小鼠并进行评价。注射CS-DS-VEGFC的小管中的精子发生明显优于注射对照的小管，并且每个小管含有更多的精原细胞和精母细胞以及沿着Ki 67和PCNA阳性细胞。此外，这些小管中的AKT和MAPK的磷酸化水平也高于对照组，表明CS-DS-VEGFC可以诱导这些途径的持续激活。结论：CS-DS-VEGFC结合输出小管注射技术是一种促进白消安致无精子症小鼠精子发生再生的可行方法。
Busulfan and other chemotherapeutic drugs used in the treatment of cancer may result in temporary or even permanent damage to spermatogenesis. During spermatogenesis, the rapidly dividing spermatogonia are highly susceptible to chemotherapy. Consequently, there is significant interest in developing an approach that could provide stimulation and regenerate spermatogenesis after chemotherapy. In a previous study, we suggested the potential application for vascular endothelial growth factor C (VEGFC) because of its key role in stimulating the proliferation of spermatogonia. However, methods to facilitate the recovery of spermatogenesis in such patients using VEGFC, or other regulatory factors, are sorely lacking because of the rapid degradation of these proteins and restrictions created by the blood-testis-barrier. To this end, we loaded VEGFC into polyanion dextran sulfate incorporated in a polycation chitosan shell to produce VEGFC sustained-release ultrafine particles (UFPs, CS-DS-VEGFC). We tested such particles in an azoospermic mouse model, created using busulfan. For each mouse, CS-DS-VEGFC was injected into the seminiferous tubules of one testis, while unloaded UFPs (CS-DS), or the VEGFC protein alone, was injected into the opposite testis as a control. All mice were sacrificed and evaluated 5 weeks later. Spermatogenesis in the tubules that were injected with CS-DS-VEGFC was clearly better than those injected with controls, and contained more spermatogonia and spermatocytes, along with Ki67 and PCNA positive-cells per tubule. In addition, the phosphorylation levels of AKT and MAPK in these tubules were also higher than in controls, indicating that CS-DS-VEGFC could induce the sustained activation of these pathways. In conclusion, CS-DS-VEGFC, combined with the efferent tubule injection technique, is a feasible approach with which to improve the regeneration of spermatogenesis in busulfan-induced azoospermic mice.