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Objective: Safrole, also called shikimol and Sassafras, is the carcinogenic and phenylpropanoid compound extracted from Sassafras tree and anise, betel, and camphor. Moreover, a high concentration of safrole can be occur in the saliva because of betel nut or areca quid chewing which a common habit observed in Southern and Southeastern Asia. Notably, macrophages are crucial phagocytic cells of the immune system. Nonetheless, to date, no evidence has been reported regarding safrole-induced proinflammatory response and the corresponding mechanism in macrophages. Materials and Methods: In the present study, the cytokines expression, NO generation, protein phosphorylation, and expression were assessed by enzyme-linked immunosorbent assay, Griess reagent, and Western blot assay, respectively. Results: In this study, we determined that safrole induces the generation of nitric oxide and proinflammatory cytokines, including tumor necrosis factor-α, interleukin-1β, and IL-6 in RAW264.7 macrophages in a concentration-dependent manner. Furthermore, inhibitor of κB (IκB) degradation was caused by safrole in a concentration-dependent manner. In addition, the phosphorylation of nuclear factor (NF)-κB and mitogen-activated protein kinase (MAPK) family, including p38 MAPK, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase, was induced by safrole began to increase at 10 μM and attained a plateau at 100 μM. Conclusion: These results indicated that safrole induces the expression of proinflammatory responses in macrophages through the NF-κB/IκB pathway and its upstream factor, MAPK family phosphorylation. [ABSTRACT FROM AUTHOR]

Astrocytes perform several functions in the brain and spinal cord. Penicillin is commonly used for establishment of experimental epilepsy models. Previous studies have demonstrated that astragaloside IV (3-o-ß-dxylopyranosyl-6-o-ß-d-glucopyranosyl-cycloastragenol; AS-IV) has comprehensive pharmacological functions on the attenuation of inflammation. In the present study, primary astrocyte cell cultures were divided into three groups: Control group, penicillin (2,500 µM) treatment group (epilepsy model), and penicillin+AS-IV (20, 40, 80 and 160 µmol/l) treatment group. The expression levels of inflammatory factors, including interleukin-1ß and tumor necrosis factor-a, were determined in the groups using western blot and reverse transcription-quantitative polymerase chain reaction analyses. The levels of members of the phosphorylated-mitogen-activated protein kinase (p-MAPK) family, including p-c-Jun N-terminal kinase 1/2, p-extracellular signal-regulated protein kinase 1/2 and p-p38, were determined using western blot analysis. Cell viability of the astrocytes was detected using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and cell proliferation was evaluated using a Cell Counting Kit-8 assay. The results revealed that AS-IV significantly suppressed the expression of penicillin-induced inflammatory factors in the astrocytes at the transcriptional and translational levels, and occurred in a dose-dependent manner. The penicillin-induced increase in the protein levels of the the p-MAPK family were notably decreased by AS-IV. In addition, the penicillin-induced downregulation of primary astrocyte viability/cell proliferation was significantly reversed by the administration of AS-IV. From these results, it was concluded that AS-IV suppressed the penicillin-induced upregulation of inflammatory factors and p-MAPK in astrocytes, ultimately attenuating epilepsy. [ABSTRACT FROM AUTHOR]