title: Triptolide Inhibited Cytotoxicity of Differentiated PC12 Cells Induced by Amyloid-Beta₂₅₋₃₅ via the Autophagy Pathway.
abstract: Evidence shows that an abnormal deposition of amyloid beta-peptide25-35 (Aβ25-35) was the primary cause of the pathogenesis of Alzheimer's disease (AD). And the elimination of Aβ25-35 is considered an important target for the treatment of AD. Triptolide (TP), isolated from Tripterygium wilfordii Hook.f. (TWHF), has been shown to possess a broad spectrum of biological profiles, including neurotrophic and neuroprotective effects. In our study investigating the effect and potential mechanism of triptolide on cytotoxicity of differentiated rat pheochromocytoma cell line (the PC12 cell line is often used as a neuronal developmental model) induced by Amyloid-Beta25-35 (Aβ25-35), we used 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) assay, flow cytometry, Western blot, and acridine orange staining to detect whether triptolide could inhibit Aβ25-35-induced cell apoptosis. We focused on the potential role of the autophagy pathway in Aβ25-35-treated differentiated PC12 cells. Our experiments show that cell viability is significantly decreased, and the apoptosis increased in Aβ25-35-treated differentiated PC12 cells. Meanwhile, Aβ25-35 treatment increased the expression of microtubule-associated protein light chain 3 II (LC3 II), which indicates an activation of autophagy. However, triptolide could protect differentiated PC12 cells against Aβ25-35-induced cytotoxicity and attenuate Aβ25-35-induced differentiated PC12 cell apoptosis. Triptolide could also suppress the level of autophagy. In order to assess the effect of autophagy on the protective effects of triptolide in differentiated PC12 cells treated with Aβ25-35, we used 3-Methyladenine (3-MA, an autophagy inhibitor) and rapamycin (an autophagy activator). MTT assay showed that 3-MA elevated cell viability compared with the Aβ25-35-treated group and rapamycin inhibits the protection of triptolide. These results suggest that triptolide will repair the neurological damage in AD caused by deposition of Aβ25-35 via the autophagy pathway, all of which may provide an exciting view of the potential application of triptolide or TWHF as a future research for AD.
authors.affiliation: School of Integrative Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China.
authors.forename: Pengjuan
authors.initials: P
authors.lastname: Xu
authors.affiliation: College of Medicine, Tianjin Key Laboratory of Tumor Microenvironment and Neurovascular Regulation, State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China.
authors.forename: Zhigui
authors.initials: Z
authors.lastname: Li
authors.affiliation: College of Life Science, Nankai University, Tianjin, 300071, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Medicine, Tianjin Key Laboratory of Tumor Microenvironment and Neurovascular Regulation, State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China.
authors.forename: Xiaochen
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Medicine, Tianjin Key Laboratory of Tumor Microenvironment and Neurovascular Regulation, State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Yang
mesh_terms: D016229:Amyloid beta-Peptides / Q000502:physiology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy; D002454:Cell Differentiation / Q000187:drug effects; D004224:Diterpenes / Q000494:pharmacology*; D004852:Epoxy Compounds / Q000494:pharmacology; D016716:PC12 Cells; D010446:Peptide Fragments / Q000502:physiology*; D010616:Phenanthrenes / Q000494:pharmacology*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D004224:Diterpenes; D004852:Epoxy Compounds; D010446:Peptide Fragments; D010616:Phenanthrenes; D017382:Reactive Oxygen Species; C074636:amyloid beta-protein (25-35); C001899:triptolide