Anti-bacterial and therapeutic effects of hydrogen mineral disinfectant treatment on Helicobacter pylori infected in vitro and in vivo model

Abstract

Background: Hypochlorous acid (HOCl) is a naturally occurring chemical that is a part of the innate immune response in humans. Recently, non-electrolyzed HOCl water has gained the attention of researchers as a new disinfecting agent owing to its high sterilization power, easy accessibility, and safety. Non-electrolyzed HOCl water was developed through mixing at a specific ratio based on hypochlorite and mineral supplements, which revealed a high oxidizing power. In this study, we investigated the effects of non-electrolyzed HOCl water on Helicobacter pylori (H. pylori) infection in C57BL/6 mice over 10 weeks. Mice were divided into three groups: normal control (NC) group supplied with purified water (PW) without infection, PW + H. pylori group supplied with PW after H. pylori infection, and HOCl + H. pylori group supplied with HOCl after H. pylori infection. Water was supplied to the mice using a mouse water bottle and was exchanged once a day. The antibacterial activity was evaluated by using optical density measurement and Z-score. Our study showed that HOCl + H. pylori group completely inhibited bacterial growth as compared to PW + H. pylori. Our heatmap results also showed HOCl + H. pylori group has excellent antibacterial activity as compared to the PW + H. pylori group. Body measurement for each group was done weekly for 10 weeks to obtain baseline data. We found that there were no significant changes among all groups. Our results showed that WBC and its differential counts, including total WBC, neutrophils, lymphocytes, monocytes, and eosinophils, were significantly inhibited in the HOCl + H. pylori group compared to those in the PW + H. pylori group. We evaluated oxidative stress markers such as reactive oxygen species (ROS) and nitric oxide (NO). In line with this, the level of ROS and calcium activity showed a significant reduction in both serum and stomach lysates in the HOCl + H. pylori group compared to those in the PW + H. pylori group. In contrast, HOCl water treatment enhanced GPx activity compared to PW treatment after H. pylori infection in both serum and stomach lysates. In addition, we evaluated anti-inflammatory stress markers such as interleukin (1β), granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-α,), etc. As a result, we found that the levels of GM-CSF, IL-1β, and TNF-α cytokine levels were significantly decreased in the HOCl + H. pylori group compared to those in the PW+ H. pylori group in the stomach lysate; however, there was no significant difference in serum. In addition, the expression levels of Bax, MMP-3, MMP-9, and TLR-4 were found to decrease after HOCl water treatment, whereas the expression level of Bcl-2 was found to be enhanced after HOCl water treatment in the stomach lysate. RT-PCR and q-PCR were performed to check the mRNA level among the three groups. We found that our HOCl + H. pylori group significantly decreased mRNA level as compared to the PW + H. pylori group. Taken together, our results suggest that drinking non-electrolyzed HOCl water has positive anti-oxidative, anti-inflammatory, and anti-apoptotic effects in H. pylori-infected mice through redox and immune regulation. Further studies are required to fully clarify the therapeutic applications of non-electrolyzed HOCl water in H. pylori-induced stomach problems.