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Abstract：
BACKGROUND: Parkinson\'s disease (PD) is the most common neurodegenerative disease worldwide. Studies have shown that insulin-like growth factor-binding protein 5 (IGFBP5) may contribute to methamphetamine-induced neurotoxicity and neuronal apoptosis in PC-12 cells and rat striatum. Here, we studied the expression and role of IGFBP5 in the 6-OHDA-toxicant model of PD.
METHODS: PC-12 and SH-SY5Y cells were exposed to 50 μm 6-OHDA for 24 h. qRT-PCR, western blotting, CCK-8 assay, EdU staining, annexin V staining, and immunofluorescence were performed to study the effects of IGFBP5-specific siRNAs. The effects of IGFBP5 on a rat 6-OHDA model of PD were confirmed by performing behavioral tests, tyrosine hydroxylase (TH) immunofluorescence staining, and western blotting.
RESULTS: In the GSE7621 dataset, IGFBP5 was highly expressed in the substantia nigra tissues of PD patients compared to healthy controls. In PC-12 and SH-SY5Y cells, IGFBP5 was upregulated following 6-OHDA exposure in a dose-dependent manner. Silencing of IGFBP5 promoted PC-12 and SH-SY5Y proliferation and inhibited apoptosis under 6-OHDA stimulation. Silencing of IGFBP5 relieved 6-OHDA-induced TH-positive neuron loss. Hedgehog signaling pathway was predicted as a downstream signaling pathway of IGFBP5. Negative regulation between IGFBP5 and sonic hedgehog (SHH) signaling pathway was confirmed in vitro. The effects of IGFBP5 silencing on SH-SY5Y cells were partially reversed using cyclopamine, a direct inhibitor of the SHH signaling pathway. In addition, silencing of IGFBP5 attenuated motor deficits and neuronal damage in 6-OHDA-induced PD rats.
CONCLUSIONS: Elevated IGFBP5 expression may be involved in 6-OHDA-induced neurotoxicity through regulation of the SHH signaling pathway.
METHODS: PC-12 and SH-SY5Y cells were exposed to 50 μ
RESULTS: In the GSE7621 dataset, IGFBP5 was highly expressed in the substantia nigra tissues of PD patients compared to healthy controls. In PC-12 and SH-SY5Y cells, IGFBP5 was upregulated following 6-OHDA exposure in a dose-dependent manner. Silencing of IGFBP5 promoted PC-12 and SH-SY5Y proliferation and inhibited apoptosis under 6-OHDA stimulation. Silencing of IGFBP5 relieved 6-OHDA-induced TH-positive neuron loss. Hedgehog signaling pathway was predicted as a downstream signaling pathway of IGFBP5. Negative regulation between IGFBP5 and sonic hedgehog (SHH) signaling pathway was confirmed in vitro. The effects of IGFBP5 silencing on SH-SY5Y cells were partially reversed using cyclopamine, a direct inhibitor of the SHH signaling pathway. In addition, silencing of IGFBP5 attenuated motor deficits and neuronal damage in 6-OHDA-induced PD rats.
CONCLUSIONS: Elevated IGFBP5 expression may be involved in 6-OHDA-induced neurotoxicity through regulation of the SHH signaling pathway.
摘要：
背景：帕金森病（PD）是世界范围内最常见的神经退行性疾病。研究表明，胰岛素样生长因子结合蛋白5（IGFBP5）可能有助于甲基苯丙胺诱导的PC-12细胞和大鼠纹状体的神经毒性和神经元凋亡。这里,我们研究了IGFBP5在PD的6-OHDA毒性模型中的表达和作用。
方法：将PC-12和SH-SY5Y细胞暴露于50μM6-OHDA中24小时。西方印迹,CCK-8测定，EdU染色,膜联蛋白V染色，并进行免疫荧光以研究IGFBP5特异性siRNA的作用。IGFBP5对PD大鼠6-OHDA模型的影响通过行为测试得到证实，酪氨酸羟化酶（TH）免疫荧光染色，和西方印迹。
结果：在GSE7621数据集中，与健康对照相比,IGFBP5在PD患者的黑质组织中高表达。在PC-12和SH-SY5Y单元格中，IGFBP5在6-OHDA暴露后以剂量依赖性方式上调。在6-OHDA刺激下，IGFBP5的沉默促进PC-12和SH-SY5Y的增殖，同时抑制细胞凋亡。IGFBP5的沉默缓解了6-OHDA诱导的TH阳性神经元丢失。Hedgehog信号通路被预测为IGFBP5的下游信号通路。在体外证实了IGFBP5和Sonichedgehog（SHH）信号通路之间的负调节。使用环巴胺部分逆转IGFBP5沉默对SH-SY5Y细胞的影响，SHH信号通路的直接抑制剂。此外,IGFBP5的沉默减轻了6-OHDA诱导的PD大鼠的运动缺陷和神经元损伤。
结论：IGFBP5表达升高可能通过调节SHH信号通路参与6-OHDA诱导的神经毒性。
方法：将PC-12和SH-SY5Y细胞暴露于50μM6-OHDA中24小时。西方印迹,CCK-8测定，EdU染色,膜联蛋白V染色，并进行免疫荧光以研究IGFBP5特异性siRNA的作用。IGFBP5对PD大鼠6-OHDA模型的影响通过行为测试得到证实，酪氨酸羟化酶（TH）免疫荧光染色，和西方印迹。
结果：在GSE7621数据集中，与健康对照相比,IGFBP5在PD患者的黑质组织中高表达。在PC-12和SH-SY5Y单元格中，IGFBP5在6-OHDA暴露后以剂量依赖性方式上调。在6-OHDA刺激下，IGFBP5的沉默促进PC-12和SH-SY5Y的增殖，同时抑制细胞凋亡。IGFBP5的沉默缓解了6-OHDA诱导的TH阳性神经元丢失。Hedgehog信号通路被预测为IGFBP5的下游信号通路。在体外证实了IGFBP5和Sonichedgehog（SHH）信号通路之间的负调节。使用环巴胺部分逆转IGFBP5沉默对SH-SY5Y细胞的影响，SHH信号通路的直接抑制剂。此外,IGFBP5的沉默减轻了6-OHDA诱导的PD大鼠的运动缺陷和神经元损伤。
结论：IGFBP5表达升高可能通过调节SHH信号通路参与6-OHDA诱导的神经毒性。
