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Astrotourism is considered to be a modern form of ecotourism. The main resource for astrotourism is a high-quality night sky, but this is very sensitive to natural as well as anthropogenic factors; for example, land utilization and expansion of urban areas often cause the negative effect of light pollution. The aim of the study is to perform a lighting survey by night sky brightness (NSB) measurements using the sky quality meter (SQM-L) at 20 study sites of the Slovenské stredohorie Upland region (Slovakia) and to assess the region’s potential for astrotourism development (PAD) using a multicriteria analysis. The NSB values ranged from 19.90 (city Žiar nad Hronom at Žiarska kotlina Basin) to 21.54 mag/arcsec2 (recreation area Poľana at Poľana Mountains). At 14 out of 20 study sites, the NSB values even reached 21.2 mag/arcsec2, as recommended by the International Dark-Sky Association for dark-sky parks. Four study sites were categorized as sites with medium PAD, and sixteen with low PAD. No study site reached a high or very high PAD. The best conditions for astrotourism development are fulfilled mainly by the Poľana Mountains geographical unit. The findings can be used for sustainable astrotourism development, land management, and planning to ensure socioeconomic development, together with nature and dark-sky conservation.