PMID- 36174498 OWN - NLM STAT- MEDLINE DCOM- 20221209 LR - 20221209 IS - 1423-0313 (Electronic) IS - 0031-7012 (Linking) VI - 107 IP - 11-12 DP - 2022 TI - Effects of Rapamycin on the Expression of Redox Enzymes in Aortic Vascular Smooth Muscle Cells from Marfan Syndrome Mice. PG - 615-622 LID - 10.1159/000526624 [doi] AB - Activation of the mechanistic target of rapamycin (mTOR) pathway has been implicated in an increasing number of diseases, including Marfan syndrome (MFS), an inherited connective tissue disorder. mTOR-dependent reactive oxygen species (ROS) formation has also been suggested to play a role in aortic aneurysm formation in MFS patients. This study aimed to characterize the effects of mTOR inhibition by rapamycin on key redox enzymes and NADPH oxidases (NOX) in cultured vascular smooth muscle cells of a murine MFS model. Therefore, the influence of 5 and 20 nmol/L rapamycin solved in 0.1% (vol/vol) DMSO on glutathione peroxidases 1 (Gpx1) and 4 (Gpx4), superoxide dismutase 2 (Sod2), and catalase (Cat) mRNA and protein expression was investigated in isolated murine aortic smooth muscle cells. Rapamycin inhibited the mRNA expression of all redox enzymes by 30-50%, except Gpx1. In the same cells, the mRNA expression of the transcription factor NFE2-related factor-2 and peroxisome proliferator-activated receptor-gamma, key factors against oxidative stress, and controlling redox gene expression were also inhibited to a comparable extent under these conditions. In addition, Nox1 but not Nox4 mRNA expression was significantly inhibited by up to 40%. DMSO alone increased nearly 2-fold the redox enzyme protein expression, which was reduced considerably to basal levels by rapamycin. Proteasomal inhibition by bortezomib could not reverse the observed decrease of GPx protein content. The rapamycin-mediated decrease in GPx protein abundance was reflected in a reduced total GPx enzymatic activity. Higher rapamycin concentrations did not further decrease but led to a renewed increase in enzymatic activity despite low GPx protein concentrations. Baseline ROS formation was slightly inhibited at 13% with 5 nmol/L rapamycin and returned to baseline levels with the higher 20 nmol/L rapamycin concentration. In conclusion, this study further characterized the mechanism of action of rapamycin. It provided an insight into how rapamycin interferes with the regulation of redox homeostasis essential for ROS-dependent signaling that does not incur cellular damage. CI - (c) 2022 S. Karger AG, Basel. FAU - Mathies, Marius AU - Mathies M AD - Department of Cardiovascular Physiology, Heidelberg University, Heidelberg, Germany. FAU - Krieg, Elisa M AU - Krieg EM AD - Department of Cardiovascular Physiology, Heidelberg University, Heidelberg, Germany. FAU - Mohr, Franziska AU - Mohr F AD - Department of Cardiovascular Physiology, Heidelberg University, Heidelberg, Germany. FAU - Zaradzki, Marcin AU - Zaradzki M AD - Department of Cardiac Surgery, University Hospital Heidelberg, Heidelberg, Germany. FAU - Wagner, Andreas H AU - Wagner AH AD - Department of Cardiovascular Physiology, Heidelberg University, Heidelberg, Germany. LA - eng PT - Journal Article DEP - 20220929 PL - Switzerland TA - Pharmacology JT - Pharmacology JID - 0152016 RN - YOW8V9698H (Dimethyl Sulfoxide) RN - EC 1.6.3.- (NADPH Oxidases) RN - 0 (Reactive Oxygen Species) RN - 0 (RNA, Messenger) RN - W36ZG6FT64 (Sirolimus) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Animals MH - Mice MH - Cells, Cultured MH - Dimethyl Sulfoxide/metabolism/pharmacology MH - *Marfan Syndrome/drug therapy/genetics/metabolism MH - Muscle, Smooth, Vascular/metabolism MH - Myocytes, Smooth Muscle/metabolism MH - NADPH Oxidases/metabolism MH - Oxidation-Reduction MH - Reactive Oxygen Species/metabolism MH - RNA, Messenger/metabolism MH - Sirolimus/pharmacology/metabolism MH - TOR Serine-Threonine Kinases/metabolism OTO - NOTNLM OT - Marfan syndrome OT - Rapamycin OT - Redox enzymes OT - Smooth muscle cells EDAT- 2022/09/30 06:00 MHDA- 2022/12/07 06:00 CRDT- 2022/09/29 18:33 PHST- 2022/03/29 00:00 [received] PHST- 2022/08/09 00:00 [accepted] PHST- 2022/09/30 06:00 [pubmed] PHST- 2022/12/07 06:00 [medline] PHST- 2022/09/29 18:33 [entrez] AID - 000526624 [pii] AID - 10.1159/000526624 [doi] PST - ppublish SO - Pharmacology. 2022;107(11-12):615-622. doi: 10.1159/000526624. Epub 2022 Sep 29.