PMID- 35487038 OWN - NLM STAT- MEDLINE DCOM- 20221213 LR - 20221213 IS - 1618-095X (Electronic) IS - 0944-7113 (Linking) VI - 101 DP - 2022 Jul TI - Lignans from Eucommia ulmoides Oliver leaves exhibit neuroprotective effects via activation of the PI3K/Akt/GSK-3beta/Nrf2 signaling pathways in H(2)O(2)-treated PC-12 cells. PG - 154124 LID - S0944-7113(22)00202-1 [pii] LID - 10.1016/j.phymed.2022.154124 [doi] AB - BACKGROUND: Neuronal apoptosis and oxidative stress have the most crucial influence on neurodegenerative diseases, including Parkinson's disease. Rat adrenal pheochromocytoma cells (PC-12) induced by H(2)O(2) are one of the primary in vitro models of Parkinson's disease (PD) . Previous studies have found that E ulmoides leaf extract exerts good neuroprotective activity and has the potential to treat neurodegenerative diseases. However, the molecular pathways involved in the neuroprotective effects of its primary leaf component, lignans, have not yet been well elucidated yet. PURPOSE: This study aimed to evaluate the neuroprotective effects of lignans in E. ulmoides leaves and to explore the underlying mechanism. METHODS: Cell viability was measured using the CCK-8 assay. Apoptosis was assessed by calcein/PI staining. The release levels of ROS and LDH were assessed using a commercial assay kit. The enzyme activities of SOD and GPx were measured using kits. The establishment of the compound-target-pathway-disease network was performed using a database and computer software. Antioxidant proteins (HO-1, NQO-1, and Cat) and related regulatory proteins (Nrf2, GSK-3beta, p-GSK 3beta (Ser 9), Akt, p-Akt (Tyr326), PI3K) were detected by western blotting. Apoptosis in the zebrafish head was assessed using acridine orange (AO) staining. RESULTS: In the present study, 12 lignans were isolated and characterized from E. ulmoides leaves, including a new compound, (-)-7‑epi-pinoresinol mr1 (1). Compounds 1-12 exerted neuroprotective effects in H(2)O(2)-treated PC-12 cells by increasing cell viability, improving the enzyme activity of SOD and GPx, and reducing levels of ROS and LDH. Compared to the positive control group (25 muM hesperetin), cell viability in response to 25 muM compound 1 (78.0 +/- 0.8%) was highest, but its relative percent LDH release (20.1 +/- 2.5%) was the lowest; 25 muM compound 4 resulted in the lowest ROS release levels (101.7 +/- 2.6%) and highest SOD enzyme activity (35.9 +/- 4.2 U/mg), and the GPx enzyme activity of 25 muM compound 1 was strongest (197.6 +/- 0.6 U/mg). Next, the potential targets (PI3K, GSK-3beta) of the test compounds' antioxidant activity were identified using pharmacological network analysis. Using DAVID software for pharmacological network analysis, potential targets (PI3K, GSK-3beta, and SOD2) of 12 lignans were identified. Based on the initial screening results, biological experiments confirmed that diepoxylignans 1, 2, and 4 exerted significant neuroprotection by regulating the PI3K/AKT/GSK-3beta/Nrf2 signaling pathways, increasing protein expression of HO-1, NQO-1, and CAT, and enhancing the antioxidant enzyme activity of SOD and GPx. CONCLUSION: Our experiments first propose that the diepoxylignans from E. ulmoides leaves exert neuroprotective effects via activation of the PI3K/Akt/GSK-3beta/Nrf2 signaling pathway. These findings further indicate that lignans could be the primary components of E. ulmoides Oliver as agents for the prevention and treatment of neurodegenerative diseases. Collectively, Eucommia ulmoides leaves with important research value may be a potential candidate for traditional Chinese medicine for treating oxidative stress-related neurodegenerative diseases. CI - Copyright (c) 2022. Published by Elsevier GmbH. FAU - Han, Rui AU - Han R AD - Shaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China. FAU - Yu, Yao AU - Yu Y AD - Shaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China. FAU - Zhao, Kanghong AU - Zhao K AD - Shaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China. FAU - Wei, Jing AU - Wei J AD - College of Biology Pharmacy & Food Engineering, Shangluo University, Shangluo, Shaanxi 726000, People's Republic of China. FAU - Hui, Yuhu AU - Hui Y AD - Shaanxi Jiahe Pharmaceutical Co., Ltd. No. 7 Binhe Road, Yangling, Shaanxi 712100, People's Republic of China. Electronic address: 13571809961@139.com. FAU - Gao, Jin-Ming AU - Gao JM AD - Shaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China. Electronic address: jinminggao@nwsuaf.edu.cn. LA - eng PT - Journal Article DEP - 20220419 PL - Germany TA - Phytomedicine JT - Phytomedicine : international journal of phytotherapy and phytopharmacology JID - 9438794 RN - 0 (Antioxidants) RN - EC 2.7.11.1 (Glycogen Synthase Kinase 3 beta) RN - BBX060AN9V (Hydrogen Peroxide) RN - 0 (Lignans) RN - 0 (Neuroprotective Agents) RN - 0 (NF-E2-Related Factor 2) RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - 0 (Reactive Oxygen Species) RN - EC 1.15.1.1 (Superoxide Dismutase) SB - IM MH - Animals MH - Rats MH - Antioxidants/metabolism MH - *Eucommiaceae MH - Glycogen Synthase Kinase 3 beta/metabolism MH - Hydrogen Peroxide/pharmacology MH - *Lignans/pharmacology MH - *Neuroprotective Agents/pharmacology MH - NF-E2-Related Factor 2/metabolism MH - *Parkinson Disease MH - PC12 Cells MH - Phosphatidylinositol 3-Kinases/metabolism MH - Proto-Oncogene Proteins c-akt/metabolism MH - Reactive Oxygen Species/metabolism MH - Signal Transduction MH - Superoxide Dismutase/metabolism MH - Zebrafish/metabolism OTO - NOTNLM OT - Diepoxylignans OT - Eucommia ulmoides oliver leaves OT - Neuroprotection OT - Phytochemicals OT - Pi3k/akt/gsk-3beta/nrf2 signaling pathway EDAT- 2022/04/30 06:00 MHDA- 2022/05/19 06:00 CRDT- 2022/04/29 18:15 PHST- 2022/01/13 00:00 [received] PHST- 2022/04/08 00:00 [revised] PHST- 2022/04/17 00:00 [accepted] PHST- 2022/04/30 06:00 [pubmed] PHST- 2022/05/19 06:00 [medline] PHST- 2022/04/29 18:15 [entrez] AID - S0944-7113(22)00202-1 [pii] AID - 10.1016/j.phymed.2022.154124 [doi] PST - ppublish SO - Phytomedicine. 2022 Jul;101:154124. doi: 10.1016/j.phymed.2022.154124. Epub 2022 Apr 19.