Human Chorionic Gonadotropin Protects Vascular Endothelial Cells from Oxidative Stress by Apoptosis Inhibition, Cell Survival Signalling Activation and Mitochondrial Function Protection

Abstract

Background/Aim: Previous reports have made it hypothetically possible that human chorionic gonadotropin (hCG) could protect against the onset of pregnancy-related pathological conditions by acting as an antioxidant. In the present study we planned to examine the effects of hCG against oxidative stress in human umbilical vein endothelial cells (HUVEC). Methods: HUVEC were subjected to peroxidation by hydrogen peroxide. The modulation of nitric oxide (NO) release by hCG and its effects on cell viability, glutathione (GSH) levels, mitochondrial membrane potential and mitochondrial transition pore opening (MPTP) were examined by specific dyes. Endothelial and inducible NO synthase (eNOS and iNOS), Akt and extracellular -signal-regulated kinases 1/2 (ERK1/2) activation and markers of apoptosis were analyzed by Western Blot. Results: In HUVEC, hCG reduced NO release by modulating eNOS and iNOS. Moreover, hCG protected HUVEC against oxidative stress by preventing GSH reduction and apoptosis, by maintaining Akt and ERK1/2 activation and by keeping mitochondrial function. Conclusion: The present results have for the first time shown protective effects exerted by hCG on vascular endothelial function, which would be achieved by modulation of NO release, antioxidant and antiapoptotic actions and activation of cell survival signalling. These findings could have clinical implications in the management of pregnancy-related disorders.

Modulation of Oxidative Stress by 17 β-Estradiol and Genistein in Human Hepatic Cell Lines In Vitro

“…Control cells were treated with DMEM 0% FBS and phenol red only. Cell viability was examined by using the 1% 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT; Life Technologies Italia, Monza, Italy) dye, as previously described [13-15]. Huh7.5 and LX-2 were treated with 60 min menadione (Sigma) alone or in the presence of genistein (10 pM; 100 nM; 1 µM) and 17 β estradiol (10 pM; 10 nM; 100 nM) given alone or 30 min before menadione.…”

“…Cells were stimulated as described for cell viability. After stimulations, the medium of cells was removed and incubated with 5,51,6,61-tetrachloro-1,11,3,31 tetraethylbenzimidazolyl carbocyanine iodide 1X diluted in Assay Buffer 1X for 15 min at 37°C in an incubator following the manufacturer's instruction and as previously performed (Invitrogen, Life Technologies Europe BV, Monza, Italy) [13-15]. The red (excitation 550 nm/emission 600 nm) and green (excitation 485 nm/emission 535 nm) fluorescence was measured using a fluorescence plate reader (BS1000 Spectra Count).…”

“…In Huh7.5 and LX-2 treated with genistein (10 pM; 100 nM; 1 µM) and 17 β estradiol (10 pM; 10 nM; 100 nM), as described for cell viability and mitochondrial membrane potential measurement, the content of GSH was determined by using a commercial kit according to the manufacturer’s instructions (Cayman, Ann Arbor, Michigan, USA) [13-15]. Briefly, 4 × 10 5 cells were lysed on ice with 2 ml of iced-cold Glutathione Assay Buffer and a rubber policeman.…”

“…At the end of stimulations, Huh7.5 and LX-2 were lysed in iced-Ripa-buffer supplemented with 1:200 sodium orthovanadate and 1:100 protease inhibitors cocktail and phenylmethanesulfonyl fluoride (1:100; Sigma). The extract proteins were quantified through bicinchoninic acid protein (Pierce, Rockford, IL, USA) and used for electrophoresis and immunoblotting studies [13-15]. …”

Qiliqiangxin improves oxidized low-density lipoprotein-induced human coronary artery endothelial cells injury

“…The excessive caspase-3 activity induced cell apoptosis in various types of cells [11][12][13]. Ox-LDL induced cells apoptosis via eliciting the caspase-3 activity [14][15][16][17][18].…”