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.
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。