The Hedgehog signaling pathway plays a principal role during embryonic development, tissue regeneration and carcinogenesis in various adult tissues. Although hedgehog signaling is important in gastric carcinogenesis, its role in Helicobacter pylori-associated gastritis is unclear. The aim of our study was to examine Sonic Hedgehog (Shh) signaling pathway in response to H. pylori infection. Thirty-one formalin-fixed, paraffin-embedded tissue specimens of chronic gastritis were retrieved from archival material. The immunoexpression of Shh, Smoothened (Smo) and Glioblastoma transcription factor 2 (Gli2) were detected using the immunohistochemical method. Sonic Hedgehog protein was expressed in H. pylori-positive and H. pylori-negative groups of patients. The immunoexpression of Shh, Smo and Gli2 proteins was lower in H. pylori-positive group compared to H. pylori-negative group, however only the differences in Shh and Smo immunoexpression were statistically significant. The immunoexpression of Shh was significantly correlated with the immunoexpression of Smo in both tested groups (p < 0.001, p < 0.02, respectively). No statistically significant correlation was found between Shh and Gli2 among H. pylori-positive and H. pylori-negative groups. The above findings support the hypothesis of the involvement of Shh signaling pathway in H. Pylori-associated gastritis.