Alarelin Acetate is a synthetic GnRH agonist which can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.

Cell viability significantly decreased when exposed to alarelin, as compared to its absence. The peak stimulatory effect on cell viability occurred at a concentration of 10^-5 M, with alarelin demonstrating dose-dependent activity[1].

Alarelin effectively suppresses gastric acid secretion in rats through direct interaction with parietal cells and by impairing vagal function [2]. Additionally, Alarelin significantly elevates the ratio of G1 phase while reducing the S phase ratio in rat gastric smooth muscle cells (GSMC) [1].

The cells are trypsinized in a solution of 2.5 g/L trypsin and seeded in a 96-well plate. After the cells are grown for 24 h to approximately 800 g/L subconfluent state, 0.1 mL medium containing 2.5% calf serum and various concentrations (0.001, 0.1, 10 μM) of alarelin is added to each well, respectively, and incubated for 24 h in a CO2 incubator. Each concentration is tested in at least 12 wells. Briefly, 15 μL of MTT solution is added to each well and incubated for 4 h. Then, the medium and MTT are removed and 150 μL of DMSO is added to each well and shaken for 10 min to dissolve the crystal. The OD is determined at 490 nm using an ELISA reader[1].