Thematic Excellence Program (Semmelweis University)(2020-4.1.1.-TKP2020) Támogató:
Innovációs és Technológiai Minisztérium
(K_21-139105) Támogató: NKFIA
(VEKOP-2.3.3-15-2016-00016) Támogató: Hungarian National Research, Development and
Innovation Office
Nemzeti szívprogram(NVKP_16-1–2016-0017) Támogató: NKFIH
(STIA-KFI-2019)
(ÚNKP-18- 4-E-17)
(ÚNKP-19-4-SE-94)
(Bolyai János Kutatási Ösztöndíj) Támogató: MTA
(EFOP-3.6.3-VEKOP-16-2017-00009)
Szakterületek:
Biokémia és molekuláris biológia
Kémiai tudományok
Helium inhalation induces cardioprotection against ischemia/reperfusion injury, the
cellular mechanism of which remains not fully elucidated. Extracellular vesicles (EVs)
are cell-derived, nano-sized membrane vesicles which play a role in cardioprotective
mechanisms, but their function in helium conditioning (HeC) has not been studied so
far. We hypothesized that HeC induces fibroblast-mediated cardioprotection via EVs.
We isolated neonatal rat cardiac fibroblasts (NRCFs) and exposed them to glucose deprivation
and HeC rendered by four cycles of 95% helium + 5% CO2 for 1 h, followed by 1 h under
normoxic condition. After 40 h of HeC, NRCF activation was analyzed with a Western
blot (WB) and migration assay. From the cell supernatant, medium extracellular vesicles
(mEVs) were isolated with differential centrifugation and analyzed with WB and nanoparticle
tracking analysis. The supernatant from HeC-treated NRCFs was transferred to naïve
NRCFs or immortalized human umbilical vein endothelial cells (HUVEC-TERT2), and a
migration and angiogenesis assay was performed. We found that HeC accelerated the
migration of NRCFs and did not increase the expression of fibroblast activation markers.
HeC tended to decrease mEV secretion of NRCFs, but the supernatant of HeC or the control
NRCFs did not accelerate the migration of naïve NRCFs or affect the angiogenic potential
of HUVEC-TERT2. In conclusion, HeC may contribute to cardioprotection by increasing
fibroblast migration but not by releasing protective mEVs or soluble factors from
cardiac fibroblasts.