PMID- 30315526 OWN - NLM STAT- MEDLINE DCOM- 20190603 LR - 20211204 IS - 1573-2576 (Electronic) IS - 0360-3997 (Print) IS - 0360-3997 (Linking) VI - 42 IP - 1 DP - 2019 Feb TI - Catalpol Inhibits Homocysteine-induced Oxidation and Inflammation via Inhibiting Nox4/NF-kappaB and GRP78/PERK Pathways in Human Aorta Endothelial Cells. PG - 64-80 LID - 10.1007/s10753-018-0873-9 [doi] AB - Hyperhomocysteinemia (HHCY) has been recognized as an independent risk factor for atherosclerosis and plays a vital role in the development of atherosclerosis. Catalpol, an iridoid glucoside extracted from the root of Rehmannia glutinosa, can produce anti-inflammatory, anti-oxidant, anti-tumor, and dopaminergic neurons protecting effects. This study aimed to determine the protecting effects of catalpol against homocysteine (HCY)-induced injuries in human aortic endothelial cells (HAECs) and uncover the underlying mechanisms: 1. HAECs were cultured with different concentrations of HCY (3 mM) and catalpol (7.5 muMu, 15 muMu, 30 muMu) for 24 h. (1) The level of MDA and GSH as well as LDH release was measured with colorimetric assay. (2) Reactive oxygen species (ROS) were detected by flow cytometry analysis. (3) Western blotting analysis was performed to detect the expression of Nox4, p22(phox), ICAM-1, MCP-1, VCAM-1, IkappaB, nucleus p65, p65 phosphorylation, caspase-3, -9, bax, bcl-2, and ER stress-related proteins. (4) The expressions of CHOP, ATF4 were measured by qRT-PCR. (5) Mitochondrial membrane potential in HCY-treated HAECs was measured by rhodamine 123 staining, and the samples were observed by confocal laser scanning microscopy. 2. DPI, PDTC, and TUDCA were used to determine the interaction among Nox4/ROS, NF-kappaB, and endoplasmic reticulum stress. 3. TUDCA or Nox4 siRNA were used to investigate whether the effect of catalpol inhibiting the over-production of ROS were associated with inhibiting ER stress and Nox4 expression. Catalpol significantly suppressed LDH release, MDA level, and the reduction of GSH. Catalpol reduced HCY-stimulated ROS over-generation, inhibited the NF-kappaB transcriptional activation as well as the protein over-expressions of Nox4, ICAM-1, VCAM-1, and MCP-1. Catalpol elevated bcl-2 protein expression and reduced bax, caspase-3, -9 protein expressions in the HCY-treated HAECs. Simultaneously, catalpol could also inhibit the activation of ER stress-associated sensors GRP78, IRE1alpha, ATF6, P-PERK, P-eIF2alpha, CHOP, and ATF4 induced by HCY. In addition, the extent of catalpol inhibiting ROS over-generation and NF-kappaB signaling pathway was reduced after inhibiting Nox4 or ER stress with DPI or TUDCA. The inhibitor of NF-kappaB PDTC also reduced the effects of catalpol inhibiting the expressions of Nox4 and GRP78. Furthermore, the effect of catalpol inhibiting the over-generation of ROS was reduced by Nox4 siRNA. Catalpol could ameliorate HCY-induced oxidation, cells apoptosis and inflammation in HAECs possibly by inhibiting Nox4/NF-kappaB and ER stress. FAU - Hu, Huimin AU - Hu H AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Wang, Changyuan AU - Wang C AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Jin, Yue AU - Jin Y AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Meng, Qiang AU - Meng Q AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Liu, Qi AU - Liu Q AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Liu, Zhihao AU - Liu Z AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Liu, Kexin AU - Liu K AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. FAU - Liu, Xiaoyu AU - Liu X AD - Department of Traditional Chinese Medicine, The Second Affiliated Hospital of Dalian Medical University, 467 Zhongshan Road, Shahekou District, Dalian, 116027, China. liuxiaoyu528@163.com. FAU - Sun, Huijun AU - Sun H AD - Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China. sunhuijun@dmu.edu.cn. LA - eng GR - 81273508/National Natural Science Foundation of China/ GR - 17Z2002/Medical Research Project of Dalian municipal commission of Health and Family Planning/ PT - Journal Article PL - United States TA - Inflammation JT - Inflammation JID - 7600105 RN - 0 (Endoplasmic Reticulum Chaperone BiP) RN - 0 (HSPA5 protein, human) RN - 0 (Heat-Shock Proteins) RN - 0 (Iridoid Glucosides) RN - 0 (NF-kappa B) RN - 0 (Reactive Oxygen Species) RN - 0LVT1QZ0BA (Homocysteine) RN - 2415-24-9 (catalpol) RN - EC 1.6.3.- (NADPH Oxidase 4) RN - EC 1.6.3.- (NOX4 protein, human) RN - EC 2.7.11.1 (EIF2AK3 protein, human) RN - EC 2.7.11.1 (eIF-2 Kinase) SB - IM MH - Aorta/*cytology MH - Cells, Cultured MH - Dose-Response Relationship, Drug MH - Endoplasmic Reticulum Chaperone BiP MH - Endoplasmic Reticulum Stress/drug effects MH - Endothelial Cells/drug effects/*metabolism MH - Heat-Shock Proteins/antagonists & inhibitors/metabolism MH - Homocysteine/*adverse effects MH - Humans MH - Inflammation/chemically induced/*prevention & control MH - Iridoid Glucosides/*pharmacology/therapeutic use MH - NADPH Oxidase 4/*antagonists & inhibitors/metabolism MH - NF-kappa B/antagonists & inhibitors/metabolism MH - Reactive Oxygen Species/*metabolism MH - eIF-2 Kinase/antagonists & inhibitors/metabolism PMC - PMC6394570 OTO - NOTNLM OT - ER stress OT - HAECs OT - NF-kappaB OT - Nox4 OT - catalpol OT - homocysteine COIS- The authors report no declarations of interest. EDAT- 2018/10/14 06:00 MHDA- 2019/06/04 06:00 PMCR- 2018/10/12 CRDT- 2018/10/14 06:00 PHST- 2018/10/14 06:00 [pubmed] PHST- 2019/06/04 06:00 [medline] PHST- 2018/10/14 06:00 [entrez] PHST- 2018/10/12 00:00 [pmc-release] AID - 10.1007/s10753-018-0873-9 [pii] AID - 873 [pii] AID - 10.1007/s10753-018-0873-9 [doi] PST - ppublish SO - Inflammation. 2019 Feb;42(1):64-80. doi: 10.1007/s10753-018-0873-9.