Novel evidence of ghrelin and growth hormone segretagogue receptor expression by human ocular tissues

Fonso, Alessia Di; Ghinassi, Barbara; Izzicupo, Pascal; Zappacosta, Roberta; Liberatore, Marcella; Gallenga, Carla Enrica; D’Amico, Maria Angela; Gallenga, P. E.; Baldassarre, Angela Di

doi:10.1016/j.regpep.2014.04.005

Cited by 7 publications

(5 citation statements)

References 27 publications

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“…For the immunoblot analysis, cells were lysed in RIPA homogenization buffer, electrophoresed in SDS–PAGE gradient 4–12% and transferred onto membranes that were then hybridized with anti- cTnT (1:500), anti-α-MHC (1:500), anti-α-SA (1:200), anti-Cx43 (1:200), anti-CACNA1C (1:1000), anti-SERCA2 (1:1000), anti-MYL7 (1:500), anti-IRX4 (1:100), anti-β 1 or anti-β 2 adrenoceptor (β 1 or β 2 AR, 1:100, Santa Cruz Technologies), anti-Nav1.5 (1:1000, Abcam), anti-KCNQ1 (1:500 ThermoFisher), anti-Kir2.1 (1:200 R&D System) or anti-G6PDH antibodies (1:1000; Santa Cruz Technologies), followed by the appropriate HRP-conjugated secondary antibodies (1:1000; Santa Cruz Technologies), as previously reported 47 . Protein expression was detected with an enhanced chemiluminescence kit (Amersham, Little Chalfont, UK) and quantified by densitometry with ImageJ Software (Supplemental Fig.…”

Section: Methodsmentioning

confidence: 99%

Cardiomyocytes Derived from Human CardiopoieticAmniotic Fluids

Baldassarre

D’Amico

Izzicupo

et al. 2018

Sci Rep

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Human amniotic fluid (hAF) cells share characteristics of both embryonic and adult stem cells. They proliferate rapidly and can differentiate into cells of all embryonic germ layers but do not form teratomas. Embryoid-bodies obtained from hAF have cardiac differentiation potential, but terminal differentiation to cardiomyocytes (CMs) has not yet been described. Our purpose was to promote cardiac differentiation in hAFcells. Cells were exposed to inducing factors for up to 15 days. Only the subset of hAF cells expressing the multipotency markers SSEA4, OCT4 and CD90 (CardiopoieticAF cells) responded to the differentiation process by increasing the expression of the cardiac transcription factors Nkx2.5 and GATA4, sarcomeric proteins (cTnT, α-MHC, α-SA), Connexin43 and atrial and ventricular markers. Furthermore, differentiated cells were positive for the calcium pumps CACNA1C and SERCA2a, with approximately 30% of CardiopoieticAF-derived CM-like cells responding to caffeine or adrenergic stimulation. Some spontaneous rare beating foci were also observed. In conclusion, we demonstrated that CardiopoieticAF cells might differentiate toward the cardiac lineage giving rise to CM-like cells characterized by several cardiac-specific molecular, structural, and functional properties.

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Section: Methodsmentioning

confidence: 99%

Cardiomyocytes Derived from Human CardiopoieticAmniotic Fluids

Baldassarre

D’Amico

Izzicupo

et al. 2018

Sci Rep

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“…We assumed that another reason why its level of production remains preserved, despite the decline of ganglion cells, which are the main source of GH, is probably the existence of another GH secretion source in the eye. As a matter of fact, positive findings of GH immunoreactivity were found in the cells of the iris, ciliary, retinal pigment epithelium, lens, cornea, and choroid but this possibly related facts have not been investigated yet ( 49 ).…”

Section: Resultsmentioning

confidence: 99%

Oxidative stress impact on growth hormone secretion in the eye

Šarić¹,

Šarić²,

Barberić³

et al. 2015

Croat Med J

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AimTo evaluate the influence of oxidative stress on extrapituitary growth hormone (GH) secretion in the eye and to analyze the interdependence between eye and serum GH levels under normal and hypoxic conditions.MethodsPars plana vitrectomy (PPV) was performed in 32 patients with developed proliferative diabetic retinopathy (PDR) and 49 non-diabetic controls, both of whom required this procedure as part of their regular treatment in the period from April 2013 to December 2014. During PPV, vitreous samples were taken and blood was simultaneously collected from the cubital vein. GH levels in serum and vitreous samples were measured by electrochemical luminescence assay. Oxidative stress was measured by enzyme-linked immunosorbent assay of advanced oxidation protein products (AOPP) and lipid hydroperoxide (LPO) in serum and vitreous.ResultsSerum AOPP levels were significantly higher than vitreous levels in both groups (P < 0.001 for each group) and LPO levels were significantly higher only in PDR group (P < 0.001). There was a significant positive correlation between serum and vitreous LPO levels in PDR group (r = 0.909; P < 0.001). Serum GH levels were significantly higher than vitreous levels in both groups (P < 0.001 for each group). Serum GH levels were significantly higher in PDR group than in controls (P = 0.012). Vitreous GH values were slightly higher in PDR group, but the difference was not significant.ConclusionOur study confirms that GH production in the eye is autonomous and independent of oxidative stress or pituitary GH influence.

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“…The RT-PCR, Western blotting and immunofluorescence assay results showed that GHSR-1a is expressed in HRMECs, similar to the findings in rat retinal vascular endothelial cells [ 15 ]. Previous studies have found that ghrelin and GHSR-1a are also expressed in the pigment epithelium of the ciliary body, retinal pigment epithelium (RPE) and iris in the human eye [ 24 ], and their expression can also be detected in the aqueous humor [ 25 ], suggesting that ghrelin plays a regulatory role in the physiological function of the eye and is closely related to the occurrence of eye diseases. In this study, GHSR-1a was detected in HRMECs, and the expression of GHSR-1a in HRMECs under high-glucose conditions was significantly lower compared with the control group, indicating that the downregulation of ghrelin-GHSR-1a signaling may be related to retinal vascular injury in diabetes.…”

Section: Discussionmentioning

confidence: 99%

The ghrelin-GHSR-1a pathway inhibits high glucose-induced retinal angiogenesis in vitro by alleviating endoplasmic reticulum stress

Yao

Zhou

et al. 2022

Eye and Vis

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Background To investigate the effect of ghrelin, a brain-gut peptide hormone, on high glucose-induced retinal angiogenesis in vitro and explore its association with endoplasmic reticulum (ER) stress. Methods Human retinal microvascular endothelial cells (HRMECs) were first divided into control and high-glucose groups, and the mRNA and protein expression levels of the receptor for ghrelin [growth hormone secretin receptor 1a, (GHSR-1a)] in cells were determined. HRMECs were then treated with high glucose alone or in combination with ghrelin or siGHSR-1a, and cell viability, migration, tube formation and the expression of the ER stress-related proteins PERK, ATF4 and CHOP were detected. Finally, to clarify whether the effects of ghrelin are related to ER stress, tunicamycin, an inducer of ER stress, was used to treat HRMECs, and cell viability, cell migration, and tube formation were evaluated. Results GHSR-1a expression in HRMECs at both the mRNA and protein levels was inhibited by high-glucose treatment. Under high-glucose conditions, ghrelin promoted cell viability and inhibited migration and tube formation, which were blocked by siGHSR-1a treatment. Ghrelin inhibited the increases in the protein levels of p-PERK, ATF4 and CHOP induced by high-glucose treatment, and combination treatment with siGHSR-1a reversed this effect of ghrelin. When tunicamycin was added, the effects of ghrelin on cell viability, migration and tube formation were all weakened. Conclusions This study experimentally revealed that ghrelin can inhibit high glucose-induced retinal angiogenesis in vitro through GHSR-1a, and alleviation of ER stress may be one of the mechanisms underlying this effect.

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Novel evidence of ghrelin and growth hormone segretagogue receptor expression by human ocular tissues

Cited by 7 publications

References 27 publications

Cardiomyocytes Derived from Human CardiopoieticAmniotic Fluids

Cardiomyocytes Derived from Human CardiopoieticAmniotic Fluids

Oxidative stress impact on growth hormone secretion in the eye

The ghrelin-GHSR-1a pathway inhibits high glucose-induced retinal angiogenesis in vitro by alleviating endoplasmic reticulum stress

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