ETHNOPHARMACOLOGICAL RELEVANCE: Paeonol has often been used as an anti-inflammatory agent in gastrointestinal diseases. However, as a primary paeonol-originated herb, the effects of Cynanchum paniculatum (CP) extracts have rarely been reported in the study on gastrointestinal inflammation models. AIM OF THE STUDY: The study aims to investigate the anti-inflammatory effects of different CP extracts, including the volatile extract (CP-VE), and its aqueous extract (CP-AE), based on their different contents of paeonol. METHODS: The contents of paeonol in the extracts were detected by HPLC. The anti-inflammatory effects of CP-VE, CP-AE and their combination (CP-VA), were compared on lipopolysaccharide (LPS)-induced macrophage RAW264.7 cells, with which the primary mechanisms were investigated in MAPK/NF-κB signaling pathway. Additionally, the effects of CP-VE at different dosages were further investigated on Helicobacter pylori (Hp)-infected gastrointestinal mucosal damaged zebrafish. RESULTS: The content of paeonol in CP-VE was 93.29%, while that in CP-AE was 0.03%. The cellular study showed that the levels of IL-1β, IL-6, TNF-α and reactive oxygen species (ROS) were significantly decreased in the CP-extract groups when compared with the model group, while their levels of IL-10 and nitric oxide (NO) were significantly increased. Compared with the LPS group, p-P65, p-JNK, p-ERK and NLRP3 protein expression was decreased in the CP-extract groups with different degrees, where CP-VA exhibited the most pronounced inhibitory activities on the phosphorylation of the proteins (p <
0.001, p <
0.01). CP-AE and CP-VE demonstrated the similar inhibition of ERK and P65 phosphorylation, while CP-AE showed slightly superior inhibition of P38, and NLRP3 than CP-VE did (p <
0.05, p <
0.01). The zebrafish study indicated that the CP-VE groups showed obvious reductions in the gastrointestinal tract areas, the fluorescence intensities and gastrointestinal neutrophil counts in a dose-dependence style, when compared with the model group. The histopathological results also indicated that CP-VE showed significant effects on the pathologic structure of the intestinal mucosa. CONCLUSION: The results showed that CP-VE (40 μg/mL) and CP-AE (700 μg/mL) showed similar significantly anti-inflammatory activity on LPS-induced macrophage RAW 264.7 cells, which indicated that the activity might be not fully related with paeonol content. It is also speculated that both CP-VE and CP-AE may exert their anti-inflammatory effects via inhibiting the MAPK/NF-κB pathways. CP-extracts are expected to be potential anti-inflammatory agents from natural source in gastrointestinal inflammation-related indications, among which CP-AE need to be further investigated on its pharmacological material basis in a more comprehensive manner.