Wang Jing, Jin Dong
Department of Acupuncture, The Second Affiliated Hospital of Tianjin University of TCM No. 69 Zengchan Road, Hebei District 300211 Tianjin China
Department of Traditional Chinese Medicine, PingJin Hospital 300211 Tianjin China.
RSC Adv. 2019 Aug 13;9(43):25232-25239. doi: 10.1039/c9ra04500g. eCollection 2019 Aug 8.
Berberine plays a neuroprotective role in neurodegenerative disorders, including Alzheimer's disease (AD). However, the underlying mechanism by which berberine inhibits AD progression remains largely unclear. The AD model was established using PC12 cells after treatment of amyloid beta (Aβ). Cells were transfected with microRNA (miRNA)-107 mimic, inhibitor, zinc finger protein 217 (ZNF217) overexpression or corresponding negative controls. Cell viability, apoptosis and inflammatory cytokine secretion were measured by MTT, flow cytometry or enzyme linked immunosorbent assay, respectively. The expressions of miR-107, ZNF217 and phosphorylated tau (p-Tau) were detected by quantitative real-time polymerase chain reaction or Western blot. The association between miR-107 and ZNF217 was explored by luciferase reporter assay and RNA immunoprecipitation. Berberine attenuated Aβ-induced viability suppression in PC12 cells. Moreover, berberine inhibited the Aβ-induced increase of inflammatory cytokine expression, apoptosis and p-Tau level in PC12 cells. miR-107 expression was reduced in Aβ-treated PC12 cells and its overexpression alleviated Aβ-induced injury, which was further weakened by combination with berberine. ZNF217 was a target of miR-107 and its addition reversed miR-107-mediated inhibition of inflammatory injury, apoptosis and phosphorylation of tau. Besides, ZNF217 protein level was decreased by berberine regulating miR-107 in Aβ-treated PC12 cells. Berberine protected against Aβ-induced inflammatory injury, apoptosis and phosphorylation of tau by regulating miR-107 and ZNF217, indicating berberine as a promising neuroprotective agent for therapeutics of AD.
黄连素在包括阿尔茨海默病（AD）在内的神经退行性疾病中发挥神经保护作用。然而，黄连素抑制AD进展的潜在机制在很大程度上仍不清楚。在用淀粉样β蛋白（Aβ）处理后，使用PC12细胞建立AD模型。细胞用微小RNA（miRNA）-107模拟物、抑制剂、锌指蛋白217（ZNF217）过表达载体或相应的阴性对照进行转染。分别通过MTT法、流式细胞术或酶联免疫吸附测定法检测细胞活力、凋亡和炎性细胞因子分泌。通过定量实时聚合酶链反应或蛋白质免疫印迹法检测miR-107、ZNF217和磷酸化tau蛋白（p-Tau）的表达。通过荧光素酶报告基因检测和RNA免疫沉淀法探究miR-107与ZNF217之间的关系。黄连素减轻了Aβ诱导的PC12细胞活力抑制。此外，黄连素抑制了Aβ诱导的PC12细胞中炎性细胞因子表达增加、细胞凋亡和p-Tau水平升高。在Aβ处理的PC12细胞中miR-107表达降低，其过表达减轻了Aβ诱导的损伤，而与黄连素联合使用后这种作用进一步减弱。ZNF217是miR-107的靶标，其添加可逆转miR-107介导的对炎性损伤、细胞凋亡和tau蛋白磷酸化的抑制作用。此外，在Aβ处理的PC12细胞中，黄连素通过调节miR-107降低了ZNF217蛋白水平。黄连素通过调节miR-107和ZNF217来保护细胞免受Aβ诱导的炎性损伤、细胞凋亡和tau蛋白磷酸化，表明黄连素是一种有前景的用于AD治疗的神经保护剂。
