PMID- 28804104 OWN - NLM STAT- MEDLINE DCOM- 20190103 LR - 20190117 IS - 1347-5215 (Electronic) IS - 0918-6158 (Linking) VI - 40 IP - 10 DP - 2017 Oct 1 TI - alpha-Cyperone Inhibits PMA-Induced EPCR Shedding through PKC Pathway. PG - 1678-1685 LID - 10.1248/bpb.b17-00183 [doi] AB - alpha-Cyperone, a sesquiterpene compound represents 25.23% of the total oil and is the most abundant compound in Cyperus rotundus oil. Endothelial cell protein C receptor (EPCR) is a main member in protein C (PC) anti-coagulation system. EPCR could be shed from cell surface, and is mediated by tumor necrosis factor-alpha converting enzyme (TACE). Nothing that EPCR is a marker of vascular barrier integrity in vascular inflammatory disease and takes part in systemic inflammatory disease. In this study, we investigated whether alpha-cyperone could inhibit EPCR shedding. To observe the effect, we investigated this issue by detection the effect of alpha-cyperone on phorbol-12-myristate 13-acetate (PMA)-induced EPCR shedding in human umbilical vein endothelial cells (HUVECs). The cells were pretreated with alpha-cyperone for 12 h, and then stimulated by PMA for 1 h. The solute EPCR (sEPCR) and expression of membrane EPCR (mEPCR) were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot. The mRNA, protein level and activity of TACE were tested by quantitative (q)RT-PCR, Western blot and InnoZyme TACE activity assay kit. Furthermore, we measured the protein level of mitogen-activated protein kinase (MAPK) signaling and protein kinase C (PKC) pathway under this condition by Western blot. The results showed that alpha-cyperone could suppress PMA-induced EPCR shedding through inhibiting the expression and activity of TACE. In addition, alpha-cyperone could inhibit PKC translocation, but not have an effect on phosphorylation of c-Jun N-terminal kinase (JNK), p38 and extracellular regulated protein kinases (ERK) 1/2. Given these results, alpha-cyperone inhibits PMA-induced EPCR shedding through PKC pathway, which will provide an experimental basis for further research on alpha-cyperone. FAU - Ma, Yu AU - Ma Y AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Zhao, Yi AU - Zhao Y AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Zhang, Ran AU - Zhang R AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Liang, Xiaoxia AU - Liang X AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Yin, Zhongqiong AU - Yin Z AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Geng, Yi AU - Geng Y AD - Department of Pharmacy, Sichuan Agricultural University. FAU - Shu, Gang AU - Shu G AD - Department of Pharmacy, Sichuan Agricultural University. FAU - Song, Xu AU - Song X AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Zou, Yuanfeng AU - Zou Y AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Li, Lixia AU - Li L AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Yin, Lizi AU - Yin L AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. FAU - Yue, Guizhou AU - Yue G AD - College of Science, Sichuan Agricultural University. FAU - Li, Yinglun AU - Li Y AD - Department of Pharmacy, Sichuan Agricultural University. FAU - Ye, Gang AU - Ye G AD - Department of Pharmacy, Sichuan Agricultural University. FAU - He, Changliang AU - He C AD - Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University. LA - eng PT - Journal Article DEP - 20170811 PL - Japan TA - Biol Pharm Bull JT - Biological & pharmaceutical bulletin JID - 9311984 RN - 0 (Endothelial Protein C Receptor) RN - 0 (Indoles) RN - 0 (Maleimides) RN - 0 (Naphthalenes) RN - 0 (PROCR protein, human) RN - 56937-68-9 (phorbolol myristate acetate) RN - EC 2.7.11.13 (Protein Kinase C) RN - EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinases) RN - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases) RN - EC 3.4.24.86 (ADAM17 Protein) RN - L79H6N0V6C (bisindolylmaleimide I) RN - NI40JAQ945 (Tetradecanoylphorbol Acetate) RN - ZL24SG1C2D (alpha-cyperone) SB - IM MH - ADAM17 Protein/metabolism MH - Cell Survival/drug effects MH - Endothelial Protein C Receptor/*metabolism MH - Human Umbilical Vein Endothelial Cells MH - Humans MH - Indoles/pharmacology MH - JNK Mitogen-Activated Protein Kinases/metabolism MH - MAP Kinase Signaling System/drug effects MH - Maleimides/pharmacology MH - Mitogen-Activated Protein Kinases/metabolism MH - Naphthalenes/*pharmacology/toxicity MH - Phosphorylation MH - Protein Kinase C/*metabolism MH - Tetradecanoylphorbol Acetate/*analogs & derivatives/antagonists & inhibitors/pharmacology MH - p38 Mitogen-Activated Protein Kinases/metabolism OTO - NOTNLM OT - endothelial protein C receptor OT - protein kinase C pathway OT - shedding OT - alpha-cyperone EDAT- 2017/08/15 06:00 MHDA- 2019/01/04 06:00 CRDT- 2017/08/15 06:00 PHST- 2017/08/15 06:00 [pubmed] PHST- 2019/01/04 06:00 [medline] PHST- 2017/08/15 06:00 [entrez] AID - 10.1248/bpb.b17-00183 [doi] PST - ppublish SO - Biol Pharm Bull. 2017 Oct 1;40(10):1678-1685. doi: 10.1248/bpb.b17-00183. Epub 2017 Aug 11.