Mouse IL-4(Interleukin 4) ELISA Kit

Helicobacter pylori, a Gram-negative flagellated microaerophilic bacterium, is associated with inflammation of the stomach, duodenal and gastric ulcer disease, and gastric cancer. This bacterium infects almost half of the world's population stomachs. One of this pathogen's immunogenic genes is the flaA gene that can stimulate the host immune system. In our previous study, the immune response in pCDNA3 1(-)-flaA infused BALB/c mice against H. pylori infection was investigated using quantitative real-time PCR (q-RT-PCR). After preparation of pCDNA3 1(-)-flaA recombinant plasmid at largescale, the mice were infused in the hip muscle by a recombinant vector with or without chitosan nanoparticles. The pcDNA3 1(-) was used as the negative control. The blood and tissue specimens of each mouse were collected at different times. The expression levels of cytokine genes (including IL-2, IFN gamma, IL4) and the internal control gene were evaluated in peripheral blood cells using a q-RT-PCR method. Also, the flaA gene expression in mice muscle was measured at 15, 30, and 45 days after the last injection. In infused mice by pcDNA3 1(-) flaA, the IL-2 and IFN gamma genes were increased statistically (p <0.001) and IL4 was significantly decreased (p <0.001). Moreover, the expression of flaA gene in mice muscles was decreased by passing the time. Furthermore, the infused mice by pcDNA3 1(-)-flaA + nanoparticles showed a better immune response because of alteration in IL4 expression. Our findings in infused mice by pcDNA3 1(-)-flaA suggested that the expression level of IL-2 and IFN gamma were increased compared to the IL-4 via simulation of Th1. Also, the expression of the flaA gene in tissue samples was decreased 45 days after the last injection. Therefore, it can be concluded that the pcDNA3.1(-)-flaA recombinant vector together with chitosan nanoparticles has the ability to stimulate the immune system and it can be investigated as a cost-effective method to control the H. pylori disease in a human in further studies.

Ethnopharmacological importance: The species Urera baccifera (L.) Gaudich. ex Wedd. (Urticaceae) is native to the Americas and is distributed widely throughout Brazil, where it is known as urtiga-brava, urtiga-vermelha, or urtigao. The leaves are often used as anti-inflammatory and antirheumatic agents and for the treatment of gastric disorders. However, the pharmacological mode of action underlying the gastroprotection induced by this species has not been investigated. Aim of the study: To contribute to the knowledge of the gastroprotective mode of action of the hydroalcoholic extract of U. baccifera (HEU) leaves. Materials and methods: Antiulcerogenic effect of HEU against ethanol-induced acute gastric ulcer was evaluated in rats and mice at doses of 3-300 mg/kg. NO-synthase inhibitor (L-NAME), SH blocker (NEM), cyclooxygenase inhibitor (indomethacin) and alpha 2-adrenergic receptor antagonist yohimbine were used to evaluate the participation of cytoprotective factors in HEU gastroprotection. Moreover, the levels of reduced gluthatione (GSH) and cytokines (TNF, IL-6, IL4 and IL-10), as well as the enzymatic activity of gluthatione S-transferase (GST), myeloperoxidase (MPO), superoxide dismutase (SOD) and catalase (CAT) were measure. Moreover, the samples were analyzed histologically and the antisecretory capability of HEU were quantified using pylorus ligated rats. Results: The phytochemical analysis of HEU (UPLC/ESI-IT-MS) identified the flavonoids diosmetin and apigenin glucuronide. Furthermore, HEU decreased the occurrence of ethanol-induced ulcers at 30 and 300 mg/kg by 57% and 66%, respectively, compared with the vehicle. The gastroprotective effects were accompanied by increased GSH levels and GST and SOD activity as well as by reduced MPO activity in vivo and in vitro, revealing antioxidant effects and inhibition of neutrophil infiltration. The beneficial effects of 30 and 300 mg/kg HEU were also observed upon histological analyses. Regarding the mode of action, the gastroprotective effect of HEU was abolished by the pre-administration of L-NAME, NEM, indomethacin or yohimbine. Moreover, HEU was able to decrease the IL-6, IL-4 and IL-10 in ulcerated tissue, as well as the pepsin activity of the gastric juice in pylorus-ligated rats. Conclusion: Together, the results confirmed that the gastroprotection elicited by HEU was due reduction in oxidative damage, neutrophil migration, and peptic activity. This work validates the popular use of U. baccifera to treat gastric disorders and supports important future research for the identification of gastroprotective molecules from this species.