# _Brassica oleraea_ Pevents HCl/Ethanol-Induced Gastric Damages in Mice
## 4 Discussion
Mucus is primarily composed to mucin, a highly viscous glycosylated proteins, and the carbohydrate side chains account for more than 80% of the weight of glycoprotein in the stomach [43]. In addition, collagen is an essential extracellular matrix protein for providing the strength and sustaining (sub)mucosal and muscular integrity of the stomach [44]. Thus, the increase in Alcian blue binding of the gastric tissue as well as increases in carbohydrate and collagen of the tissue homogenates provide a direct evidence that BOE can protect from H/E-induced gastric damages by restoring mucus and sustaining mucosal integrity. It has been reported that enhanced secretion of gastric acids facilitates gastric damage by excess intake of ethanol or acids [28; 45]. Among diverse neurohumoral signaling molecules, histamine released from enterochromatin-like cells and/or neuroinflammatory cells stimulates the secretion of gastric acid in the parietal cells via H2R activation. In addition, gastrin directly activates CCK2R in the parietal cells and indirectly stimulates histamine release from the enterochromaffin-like cells [46; 47]. Activation of H2R and CCK2R leads to pumping out H\({}^{+}\) ion to the gastric lumen via H\({}^{+}\)/K\({}^{+}\)-ATPase [46]. In the present study, a significant increase in plasma histamine level was shown in H/E group, along with related increases in H2R, CCK2R, and H\({}^{+}\)/K\({}^{+}\)-ATPase mRNAs of the gastric tissue. However, H/E-mediated those changes were significantly blocked by pre-administration of BOE for 7 days, suggesting that BOE-dependent gastric protection from H/E can be attributed to the modulation of gastric acid. Histamine secreted from the mast cells and basophils contributes to increase permeability of injured tissue and to recruit diverse inflammatory cells into the damaged sites. MPO is a representative enzyme stored in the azurophilic granules of the neutrophils [48]. In conjunction with the reduction in plasma histamine, present results showed that pre-administration of three different doses of BOE significantly decreased the MPO activity in the gastric tissue, suggesting that BOE can attenuate the infiltration of neutrophil to the gastric tissue. In parallel with previous report [6], the present results also showed that BOE significantly reduced the level of gastric NF-kB (p65 subunit) mRNA induced by H/E. NF-\(\kappa\)B is a pivotal transcription factor, which accelerates pro-inflammatory response (e.g., cytokine and adhesion molecule) in a wide range of tissue including the stomach [49]. Damage-associated molecular patterns released from injured cells stimulate degradation of inhibitory \(\kappa\)B, which sequestrate NF-\(\kappa\)B in the cytoplasm, allow the NF-\(\kappa\)B to translocate into the nucleus, and lead to an increase in the transcription of target genes including COX-2, iNOS, TNF-\(\alpha\), IL-1\(\beta\), IL-6, and IL-18 [50; 51]. Although inhibition of NF-\(\kappa\)B signaling pathway by BOE may be further established by other experimental approaches (e.g., subcellular expression of NF-\(\kappa\)B and post-translational modification of NF-\(\kappa\)B signaling molecules), present results suggest that not only recruitment of inflammatory cells into the lesions but also expression of inflammatory mediators can be suppressed by three different doses of BOE through the modulation of NF-\(\kappa\)B-dependent signaling molecules. One of the most important executors in alcohol-mediated tissue damage is an oxidative stress, which is accumulated by dysregulation between endogenous antioxidants defense system and reactive oxygen (nitrogen) species [52]. Results from the present study showed that ethanol elevated the level of malondialdehydedehyde (a representative marker of lipid peroxidation) and decreased the activities of catalase and superoxide dismutase in the gastric tissue, which is parallel with previous reports that alcohol provokes gastric damage through the induction of oxidative stress [7; 8].