Based On The NF-κB/Nrf-2 Signaling Pathway,the Mechanism Of Hydrogen-rich Water In Alleviating Chronic Intestinal Inflammation In Rats Was Explored

Chronic inflammation is a major pathogenic factor for a variety of chronic diseases,which can cause damage to multiple organs of animals.Intestinal tissues are easily exposed to inflammatory environment for a long time,resulting in the destruction of intestinal barrier function and intestinal damage.Inflammatory factors enter the blood circulation and cause chronic inflammation.Hydrogen-rich water has antioxidant and anti-inflammatory effects,which can protect the intestinal barrier from the damage of inflammation.However,the protective effects of long-term drinking hydrogen-rich water on the body and intestinal tract have not been reported.Therefore,the purpose of this study was to establish a rat model of chronic inflammation to explore the protective effect of long-term drinking hydrogen-rich water on animal body and intestinal tract,as well as the protective mechanism of Nrf-2/NF-κB signaling pathway in reducing intestinal damage caused by chronic inflammation caused by hydrogen-rich water.Forty-eight healthy 8-week-old rats were selected and randomly divided into control group(CON),hydrogen-rich water group(HRW),lipopolysaccharide group(LPS)and hydrogen-rich water treatment group(HRW+LPS).The animals were fed for 8 months,during which the HRW and HRW+LPS groups drank fresh hydrogen-rich water prepared daily,and the CON and LPS groups drank distilled water.After 8 months,the rats in LPS group and HRW+LPS group were injected with LPS through tail vein for 4 weeks.After 4 weeks,the rats were killed and serum and tissue samples were collected.The contents of cytokines,MPO,SOD,MDA and ovalbumin(OVA)were detected by ELISA,the expressions of NFκB and Nrf-2 signaling pathway related proteins were detected by western blot,the expressions of tight junction protein and CD68 were detected by immunohistochemistry,and the levels of ROS were detected by immunofluorescence.The expression of related genes was detected by q-PCR,and the heart,liver,spleen,lung and ileum tissues were collected for histopathological examination.The results show that: long-term HRW consumption could slow down the weight loss of rats and protect the organ structure from the damage of inflammation.Chronic inflammation can activate NF-κB signaling pathway.Compared with CON group,NF-κB signaling pathway was significantly activated in LPS group,and the protein expression of TLR4,My D88,p-IκBα,p-p50 and p-p65 was increased,while the protein expression of IκBα was decreased.Compared with LPS group,the protein expressions of TLR4,My D88,p-p50 and p-p65 in HRW+LPS group were decreased,while the protein expressions of IκBα and p-IκBα were increased,and the same result was shown in gene expression.HRW could inhibit the activation of NF-κB signaling pathway.Compared with CON group,chronic inflammation resulted in increased contents of TNF-α,IL-1β,IL-6,IL-10,MCP-1and COX-2 in tissues and serum of LPS group and increased gene expression.Compared with LPS group,the contents of cytokines and COX-2 in tissue and serum of HRW+LPS group were decreased,and the gene expression was decreased.Macrophages gathered at the site of inflammation.Compared with CON group,the level of macrophages in LPS group was significantly increased and infiltration was serious.Macrophage levels were significantly decreased in the HRW+LPS group compared with the LPS group.Chronic inflammation also activated the Nrf-2 signaling pathway.Compared with the CON group,the protein expressions of Nrf-2,HO-1 and NQO1 in the HRW group and the HRW + LPS group were increased,while the protein expressions of HO-1 and NQO1 in the LPS group were decreased.Compared with LPS group,the protein expressions of Nrf-2,HO-1 and NQO1 in HRW + LPS group were increased.Compared with CON group,chronic inflammation resulted in increased ROS content and increased MPO content in LPS group.Compared with LPS group,the ROS and MPO contents were decreased in HRW + LPS group.Chronic inflammation could lead to significant changes in oxidative stress-related enzymes.Compared with CON group,SOD content in LPS group was lower and MDA content was higher.Compared with LPS group,SOD content was increased and MDA content was decreased in HRW +LPS group.Chronic inflammation would destroy the intestinal structure.Compared with CON group,the expression of tight junction protein in the intestinal tissue of LPS group was significantly decreased,the intestinal structure was destroyed,OVA was introduced into the blood,and the content of OVA in the blood was significantly increased.Compared with LPS group,tight junction protein expression was increased and OVA content was decreased in HRW + LPS group.The results showed that HRW inhibited the activation of NF-κB signaling pathway,reduced the contents of inflammatory factors,COX-2,MPO,ROS and MDA,alleviated the infiltration of macrophages,activated the Nrf-2 signaling pathway,significantly increased the level of SOD and tight junction protein,and finally alleviated the intestinal damage caused by chronic inflammation in rats.