Protective Effects of Hexarelin and JMV2894 in a Human Neuroblastoma Cell Line Expressing the SOD1-G93A Mutated Protein.

In vitro study examining the effects of growth hormone secretagogues on human neuroblastoma cells expressing SOD1-G93A under oxidative stress.

“In this in vitro study, we investigated potential protective effects of hexarelin and JMV2894 on human neuroblastoma cells overexpressing the SOD1-G93A mutated protein in order to ascertain whether these two GHSs could be candidates for the development of new drugs for the treatment of ALS.”

Human neuroblastoma cells (SH-SY5Y) overexpressing wild-type or SOD1-G93A mutated protein.

“Human neuroblastoma cells expressing the SOD1-G93A mutated protein (SH-SY5Y SOD1G93A cells) were incubated for 24 h with H2O2 (150 µM) in the absence, or presence, of GHS (1 µM), in order to study the protective effect of GHS against increased oxidative stress.”

Experiments were independently replicated at least three times with varying numbers of replicates per group (e.g., n=21 for viability, n=100 cells analyzed for morphology).

“The results are expressed as mean ± SEM of data obtained from 3 independent experiments. In each experiment, groups consisted of 7 replicates for each treatment (total n = 21).”

Cells were incubated for 24 hours with treatments.

“Human neuroblastoma cells expressing the SOD1-G93A mutated protein (SH-SY5Y SOD1G93A cells) were incubated for 24 h with H2O2 (150 µM) in the absence, or presence, of GHS (1 µM), in order to study the protective effect of GHS against increased oxidative stress.”

Hexarelin and JMV2894 protected cells from H2O2-induced cytotoxicity and reduced morphological changes associated with apoptosis.

“The two GHSs examined in this study, hexarelin and JMV2894, protected cells from H2O2-induced cytotoxicity by activating molecules that regulate apoptosis and promote cell survival processes.”

GHS treatment reduced the expression of pro-apoptotic markers (Bax, caspase-3) and increased anti-apoptotic markers (Bcl-2) in H2O2-treated cells.

“The levels of Bax mRNA, which were increased by H2O2 (p < 0.001), were significantly (p < 0.05) reduced by co-incubation with hexarelin or JMV2894. However, only hexarelin significantly (p < 0.05) increased Bcl-2 mRNA levels.”

GHS treatment reduced DNA damage markers (γH2AX) and modulated MAPK and Akt signaling pathways in response to oxidative stress.

“Both hexarelin and JMV2894 significantly reduced the DNA damage induced by oxidative stress: in fact, hexarelin and JMV2894 significantly blunted the γH2AX foci distribution (p < 0.001), compared to the H2O2-treatedgroup.”

The study was conducted in vitro using a cell line model, and further investigations are required to clarify molecular mechanisms and develop new GHSs for therapy.

“Further investigations are required for the following: (i) to clarify GHS molecular mechanisms of action, and (ii) to envisage the development of new GHSs that may be useful in ALS therapy.”