Helicobacter pylori is a Gram-negative, micro-aerophilic bacterium that inhabits various areas of the stomach and duodenum. It causes a chronic inflammation of the stomach lining and strongly linked to the development of duodenal and gastric ulcers and stomach cancer. It has been well confirmed that infection with H. pylori is an etiological agent of peptic ulcer, and eradication of this organism can not only make the associated gastritis regression but cure these diseases. There are several treatments of H. pylori infection, most common treatment is triple therapy, including two antibiotics and a proton pump inhibitor. However, the increasing of antibiotic resistant rate was associated with the usage of antimicrobial agents in the treatment of H. pylori infection. Thus, the complementary therapeutic methods in eradication of H. pylori were needed. Antrodia camphorata (A. camphorata) is a rare treasure in Taiwan, the fruiting body of A. camphorata is well known by named niu-chang-chih or jang-jy. The miraculous efficacy in human body had been investigated and researched by many organizations in Taiwan. In this study, thirteen compounds were isolated from A. camphorata. Among them, compound 15α-Acetyldehydrosulphurenic acid (number 3) and Antcin C (number 10) had higher inhibition effects on H. pylori-induced inflammatory responses in gastric epithelial cells and mouse macrophage RAW 264.7 cells than other compounds. In addition, these constituents also inhibited the IL-8 protein expression and NF-κB signal activation in H. pylori-infected gastric epithelial cells and it might contribute to the potential role of A. camphorata as an anti-inflammatory action. Results from our study might bring a new method for treating of H. pylori infection.