Artikel
Rapalink-1 ameliorates aging and endothelial damage caused by smoke condensate – a possible molecular target in intracranial aneurysm formation
Rapalink-1 mildert Alterung und von endothelialen Schäden durch Rauchkondensat – ein mögliches molekulares Ziel zur Behandlung von intrakraniellen Aneurysmen
Suche in Medline nach
Autoren
-
Xuanchen Li
- Universitätsklinikum Düsseldorf, Klinik für Neurochirurgie, Düsseldorf, Deutschland
-
Dilaware Khan
- Universitätsklinikum Düsseldorf, Klinik für Neurochirurgie, Düsseldorf, Deutschland
-
Huakang Zhou
- Universitätsklinikum Düsseldorf, Klinik für Neurochirurgie, Düsseldorf, Deutschland
-
Carsten Hagenbeck
- Universitätsklinikum Düsseldorf, Department of Gynecology and Obstetrics, Düsseldorf, Deutschland
-
Daniel Hänggi
- Universitätsklinikum Düsseldorf, Klinik für Neurochirurgie, Düsseldorf, Deutschland
-
Sajjad Muhammad
- Universitätsklinikum Düsseldorf, Klinik für Neurochirurgie, Düsseldorf, Deutschland
| Veröffentlicht: | 25. Mai 2022 |
|---|
Gliederung
Text
Objective: Smoking is a well-known risk factor for intracranial aneurysm formation and rupture. Smoking-mediated inflammation leading to endothelial dysfunction and damage to the arterial wall are possible mechanisms behind it. This study aims to investigate the smoke condensate (SC) mediated cellular and molecular events in human umbilical vein endothelial cells (HUVECs) and find novel drug targets to block smoking-induced signals.
Methods: HUVECs (Promocell, Heidelberg, Germany) were treated with different concentrations (20µg/ml, 40µg/ml) of SC with or without Rapalink-1 (0.5nm). SC was prepared by passing cigarette smoke through ethanol. Ethanol was evaporated and smoke condensate was dissolved in DMSO. Telomere length was quantified as telomere to single-copy gene (TS) ratio. Telomere length, the mRNAs expression, and protein level were quantified by qRT-PCR and Western blot. All experiments were performed with three biological replicates, and each biological replicate had technical triplicates. For statistical analysis, the Student t-test was applied for two groups and one-way ANOVA to compare more than two groups.
Results: Exposure to SC enhanced cellular aging (TS ratio: Control=1±0.15, SC=0.59±0.2) and increased relative mRNA expression of aging related biomarkers (P21: Control=1±0.24, SC=2.77±0.68), inflammatory cytokines (IL-8: Control=1±0.25, SC=2.02±0.18; MCP-1: Control=1±0.03, SC=5.64±0.58) and MMP2 (Control=1±0.03, SC=2.07±0.42) in HUVECs. The screening of 171 drugs revealed Rapalink-1 an mTOR pathway inhibitor, which powerfully reduced SC mediated aging (TS ratio: Control=1±0.15, SC=0.59±0.2, SC+Rapalink-1=0.7±0.03, P21: Control=1±0.24, SC=2.77±0.68, SC+Rapalink-1=1.58±0.07). The expression of inflammatory cytokines (IL-8: Control=1±0.25 SC=2.02±0.18, SC+Rapalink-1=1.59±0.08; IL-17: Control=1±0.15, SC=36±15.68, SC+Rapalink-1=5.06±1.35; MCP-1: Control=1±0.03, SC=5.64±0.58, SC+Rapalink-1=3.9±0.11), receptor TLR4 (Control=1±0.1, SC=1.8±0.2, SC+Rapalink-1=0.5±0.02) and MMP2 (Control=1±0.03, SC=2.07±0.42, SC+Rapalink-1=1.26±0.09) was reduced after rapalink-1 treatment. All values are mean±StDev and P<0.05.
Conclusion: Smoke condensate induced aging and endothelial dysfunction, which was reversed by treatment with mTOR inhibitor Rapalink-1. Rapalink-1 and the mTOR pathway could be targeted in endothelial dysfunction mediated diseases, including the formation of intracranial aneurysms.
