Social behavior is an essential need for humans and animals, including Drosophila melanogaster. Recently, there has been an increasing interest in studying the impact of social isolation, the deprivation of social connections, on behavior and brain chemistry. These studies provide valuable information about changed social interactions that are symptoms of different neuropsychiatric disorders. To investigate the relationship between social isolation and chemical alterations in the brain, we conducted a study where we assessed the concentration of neurotransmitters in D. melanogaster head after a period of one and five days of social isolation. Our LC-MS/MS analysis revealed a significant decrease in concentrations of dopamine, octopamine, glutamate and acetylcholine. Furthermore, to determine if there was a correlation between neurochemical alterations and behavioral modifications, we recorded the social interactions among a group of 12 flies in a circular arena. Python scripts were used to generate and evaluate Social Interaction Networks (SINs) based on five parameters: global efficiency, assortativity, clustering coefficient, betweenness centrality, and closeness centrality. Network analysis revealed that social isolation led to an increase in the clustering coefficient and a decrease in the closeness centrality. This suggests that flies tend to establish smaller but temporary social groups within the network. To reverse the alterations in SINs caused by isolation, we administered dopamine, octopamine and combination of both during five days of isolation. Examination of SINs characteristics of supplemented groups revealed significant variations in clustering coefficients, centrality measures, and assortativity, with noticeable impacts of dopamine and octopamine. The addition of octopamine and the combination of dopamine and octopamine greatly increased closeness centrality, suggesting that flies established more densely linked networks, like what was found in groups without isolation. Additionally, we observed that differences in the age of grouped flies significantly influence local SINs parameters, with younger flies exhibiting similar behavioral patterns as isolated flies. Our results show that social isolation has lasting neurochemical and behavioral effects, in particular the behavior in a group setting. Moreover, because supplementation with dopamine and octopamine reversed some negative effect of social isolation, supplementation with these neurotransmitters might contribute in the creation of novel therapies targeted at alleviating the negative effects of social isolation.