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目的研究SynaptotagminⅡ(简称Syt2)在RBL2H3(简称RBL)胞吐中的作用。方法利用钙离子载体和抗原分别刺激Syt2+和Syt2(分别为Syt2过表达和表达减弱的RBL),诱导其活化而分泌,检测分泌的β氨基已糖苷酶、组织蛋白酶D和5羟色胺,分析Syt2对于细胞分泌的影响。结果与对照相比,Syt2+分泌的β氨基已糖苷酶无明显区别,而Syt2分泌的β氨基已糖苷酶增加达3.5倍,结果说明Syt2的过表达对β氨基已糖苷酶分泌无影响,而Syt2表达减弱却促进β氨基已糖苷酶的分泌。与对照相比,Syt2+分泌的组织蛋白酶D减少近一半,而Syt2分泌的组织蛋白酶D增加近1倍,说明Syt2的表达量与组织蛋白酶D分泌量成负相关关系。与对照相比,Syt2+分泌的5羟色胺无明显差别,而Syt2分泌的5羟色胺有少量增加,说明Syt2的表达量对于5羟色胺分泌有轻微影响。结论Syt2主要对于RBL溶酶体胞吐起负调控作用。
Objective To study the role of Synaptotagmin Ⅱ (Syt2) in the exocytosis of RBL2H3 (RBL). Methods Syt2 + and Syt2 were stimulated by calcium ionophore and antigen, respectively, which induced the activation and secretion of Syt2. The secretion of β-aminohexosaminidase, cathepsin D and serotonin were detected. The effect of Syt2 on The effects of cell secretion. Results Compared with the control, the Syt2 + secretion of β-aminohexosaminidase no significant difference, while Syt2 secreted β-amino-hexosaminidase increased up to 3.5 times, the results show that Syt2 overexpression of β-aminohexosidase secretion had no effect, and Syt2 The diminished expression promotes the secretion of β-aminohexosidase. Compared with the control, Syt2 secretion of cathepsin D decreased by nearly half, and Syt2 secreted cathepsin D increased nearly doubled, indicating that the expression of Syt2 and cathepsin D secretion was negatively correlated. Compared with the control, serotonin secretion Syt2 no significant difference, while the secretion of serotonin Syt2 a small increase, indicating that the expression of Syt2 serotonin secretion slightly affected. Conclusion Syt2 is mainly responsible for the negative regulation of RBL lysosome exocytosis.