Department of Biological Sciences, University of North Texas, Denton, TX, USA.
Platelets. 2022 Jan 2;33(1):54-58. doi: 10.1080/09537104.2021.1882667. Epub 2021 Feb 4.
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.
在人类中，脾切除术用于治疗许多临床疾病，包括免疫性血小板减少症。然而，由于新疗法的出现，过去十年中，作为治疗方法的脾切除术的发病率显著下降。脾切除术导致感染和败血症的情况已有详细记录，但其他长期影响尚未得到很好的描述。有证据表明，先前接受过脾切除术的人可能面临血管疾病的风险增加。此外，细胞衍生的微颗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，关于脾切除术后微颗粒水平升高的体内研究有限。为了了解脾切除术的影响，我们在成年斑马鱼中制定了脾切除术方案。麻醉后，在鱼的腹侧做一个切口，在立体显微镜下切除脾脏。用镊子夹住脾脏并将其取出。切口用 Vetbond 组织胶封闭。从脾切除斑马鱼和接受假手术的斑马鱼采集的血液用多克隆抗血清针对 αIIb 进行免疫标记，然后进行流式细胞术分析。我们观察到脾切除斑马鱼中血小板及其微颗粒水平升高。最后，通过向斑马鱼静脉内注射 αIIb 抗体，我们发现血小板计数减少，表明鱼出现了类似于免疫性血小板减少症的情况，随后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治疗类似于免疫性血小板减少症的情况。
