Splenectomy in zebrafish: a new model for immune thrombocytopenia.
Splenectomy in zebrafish: a new model for immune thrombocytopenia.
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DOI：
10.1080/09537104.2021.1882667
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发表时间：
2022-01-02
期刊：
影响因子：
3.3
通讯作者：
Jagadeeswaran P
中科院分区：
文献类型：
--
作者：
Radhakrishnan UP;Al Qaryoute A;Raman R;Jagadeeswaran P
在人类中，进行脾切除术是为了治疗许多临床疾病，包括免疫性血小板减少症。然而，由于新疗法的出现，在过去十年中，用于治疗免疫性血小板减少症的脾切除术的发生率显着下降。脾切除术导致的感染和败血症已有充分记录，但其他长期影响尚不清楚。有证据表明，曾接受过脾切除术的人患血管疾病的风险可能会增加。此外，细胞来源的微粒水平升高似乎会增加血栓形成和心血管疾病的风险。然而，关于脾切除术后微粒水平增加的体内研究是有限的。为了了解脾切除术的影响，我们制定了成年斑马鱼脾切除术的方案。麻醉后，在鱼腹侧切开切口后，在立体显微镜下取出脾脏。用镊子拉动取出脾脏。用 Vetbond 组织胶封闭切口。从脾切除的斑马鱼和接受假手术的斑马鱼收集的血液用针对 αIIb 的多克隆抗血清进行免疫标记，然后进行流式细胞术。我们观察到脾切除的斑马鱼中血小板及其微粒水平升高。最后，通过向斑马鱼静脉注射αIIb抗体，我们发现血小板计数减少，这表明斑马鱼出现了免疫性血小板减少症样病症，然后通过脾切除术逆转了这种情况。总之，这里开发的模型应该有助于研究脾切除引起的分子变化。此外，斑马鱼将有助于模拟免疫性血小板减少症等病症的治疗。
In humans, splenectomy is performed to treat many clinical disorders, including immune thrombocytopenia. However, the incidence of splenectomies for immune thrombocytopenia as a therapeutic has significantly declined over the past decade due to the availability of new therapies. Infection and sepsis as a result of splenectomies are well documented, but other long-term effects are not well characterized. Evidence suggests that persons who have had a prior splenectomy may be at an increased risk of vascular conditions. Also, elevated levels of cell-derived microparticles appear to contribute to an increased risk of thrombosis and cardiovascular disease. However, in vivo studies on the increased levels of microparticles following splenectomy are limited. In order to understand the effects of splenectomies, we developed a protocol for splenectomy in adult zebrafish. After anesthesia, the spleen was removed under a stereomicroscope after making an incision on the ventral side of the fish. The spleen was removed by pulling with forceps. The incision was closed by Vetbond tissue glue. Blood collected from both splenectomized zebrafish and those that underwent sham surgeries was immunolabeled with polyclonal antisera against αIIb, followed by flow cytometry. We observed elevated levels of thrombocytes and their microparticles in splenectomized zebrafish. Finally, by injecting αIIb antibody intravenously into zebrafish, we found the thrombocyte counts decreased, suggesting the fish developed immune thrombocytopenia like conditions, which were then reversed by splenectomy. In summary, the model developed here should be useful to study molecular changes due to splenectomy. Also, the zebrafish will be useful in modeling treatment of immune thrombocytopenia like conditions.
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