Methane Alleviates Inflammation and Apoptosis of Dextran Sulfate Sodium-Induced Inflammatory Bowel Diseases by Inhibiting Toll-Like Receptor 4 (TLR4)/Myeloid Differentiation Factor 88 (MyD88)/Nuclear Translocation of Nuclear Factor-κB (NF-κB) and Endoplasmic Reticulum Stress Pathways in Mice.

BACKGROUND Inflammatory bowel diseases (IBDs) are chronic idiopathic diseases with increased occurrence and recurrence rates. The aim of this study was to explore whether methane-rich saline (MRS) would be beneficial to IBD. MATERIAL AND METHODS Dextran sulfate sodium (DSS) was utilized to establish an IBD model. Male C57BL/6J mice were randomly grouped as follows: the control group, the DSS+NS group, the DSS+5-ASA group, the DSS+MRS (1) and DSS+MRS (10) groups. Seven days after model induction, blood and colon tissues were collected to assess the treatment effects. RESULTS The DSS+MRS (10) group showed obviously reduced weight loss, disease activity index, and spleen index. The isolated colon samples had a notably longer length, less thickness and weight, and better macroscopic score with MRS treatment compared with the DSS+NS group. Additionally, assessment of morphological impairment revealed a milder and lower microscopic score in the DSS+MRS (10) group, consistent with the myeloperoxidase (MPO) results. The inflammation-related molecules levels were dramatically reduced by MRS. MRS also significantly reduced oxidative stress related proteins. In addition, apoptotic cells were visually decreased in the DSS+MRS (10) group, in which the pro-apoptotic molecules Bax and cleaved caspase-3 were reduced, whereas the level of Bcl-2 was increased. Furthermore, MRS markedly decreased the TLR4, MyD88, p-NF-kappaB p65, p-IKKalphaß, and p-IkappaBalpha, and increased IL-10, p-JAK1, and p-STAT3 expression levels. Proteins involved in endoplasmic reticulum stress (ERS) were also notably reduced under MRS treatment. CONCLUSIONS MRS exerts protective effects on DSS-induced IBD via inhibiting inflammatory reaction, promoting anti-inflammatory capacity, suppressing oxidative stress, and ameliorating apoptosis.

Emodin induces human T cell apoptosis in vitro by ROS-mediated endoplasmic reticulum stress and mitochondrial dysfunction.

AIM: To elucidate the molecular mechanisms underlying the immunosuppressive effects of emodin isolated from Rheum palmatum L. METHODS: Human T cells were isolated from the peripheral venous blood of 10 healthy adult donors. Cell viability was analyzed with MTT assay. AO/EB and Annexin V/PI staining and DNA damage assay were used to detect cell apoptosis. Fluorescence staining was used to detect the levels of ROS, the mitochondrial membrane potential and intracellular Ca(2+). Colorimetry was used to detect the levels of MDA and total SOD and GSH/GSSG ratio. The expression and activity of caspase-3, -4, and -9 were detected with Western blotting and a fluorometric assay. Western blotting was also used to detect the expression of Bcl-2, Bax, cytochrome C, and endoplasmic reticulum (ER) markers. RESULTS: Emodin (1, 10, and 100 µmol/L) inhibited the growth of human T cells and induced apoptosis in dose- and time dependent manners. Emodin triggered ER stress and significantly elevated intracellular free Ca(2+) in human T cells. It also disrupted mitochondrial membrane potential, and increased cytosolic level of cytochrome C, and the levels of activated cleavage fragments of caspase-3, -4, and -9 in human T cells. Furthermore, emodin significantly increased the levels of ROS and MDA, inhibited both SOD level and GSH/GSSG ratio in human T cells, whereas co-incubation with the ROS scavenger N-acetylcysteine (NAC, 20 µmol/L) almost completely blocked emodin-induced ER stress and mitochondrial dysfunction in human T cells, and decreased the caspase cascade-mediated apoptosis. CONCLUSION: Emodin exerts immunosuppressive actions at least partly by inducing apoptosis of human T cells, which is triggered by ROS-mediated ER stress and mitochondrial dysfunction.

Tanshinone IIA May Inhibit Gastric Cancer via Affecting the Intestinal Microbiome.

Background: Gastric cancer (GC) belongs to a type of the most deadly cancer in the world, and the incidence rate of GC will increase in the coming decades. Tanshinone IIA (Tan IIA) is an active component that separated from Danshen. Tan IIA may also exert its therapeutic effects in disease with intestinal dysbacteriosis, at least partially, via regulating the intestinal microbiome. Nevertheless, it is obscure whether Tanshinone IIA affects the intestinal dysbacteriosis and plays antitumor roles. This research was designed to explore Tanshinone IIA potential on the intestinal dysbacteriosis of GC xenograft mice. Methods: Mouse xenograft GC tumor models were built and treated by Tan IIA. The tumor growth as well as microbiome in the intestinal were compared. Western blot was used to detect the phosphorylation of the NF-κB and expressions of the downstream cytokines IL-6 and IL-1ß. Results: Microbiome in the intestinal was changed in xenograft tumor mice in comparison with the control mice. What is more, Tan IIA could influence the microbiome in the intestinal of the tumor mice. Tan IIA hinders the growth of xenograft tumor and change the microbiome in the intestinal, but intestinal dysbacteriosis condition partially blocked Tan IIA-stimulated antitumor effects. In addition, intestinal dysbacteriosis abrogated Tan IIA-stimulated decrease in the NF-κB signaling in xenograft tumor mice. Conclusions: Tanshinone IIA may inhibit GC tumor growth via affecting the intestinal microbiome through regulating the NF-κB signaling.

Mesenteric lymphatic ducts ligation decreases the degree of gut-induced lung injury in a portal vein occlusion and reperfusion canine model.

BACKGROUND: Whether the mesenteric lymphatic system could serve as a route of transport by which gut-derived inflammatory mediators contribute to the induction of remote organ injuries is uncertain. We therefore made a gut-induced lung injury canine model by portal vein occlusion and reperfusion (PV O/R) and studied the role of mesenteric lymphatic ducts ligation (ML) to gut-induced lung injury with this model. MATERIAL AND METHODS: Eighteen mongrel dogs were divided into control, PV O/R, and PV O/R + ML groups. Cytokines and endotoxin levels in the portal vein and lymph from thoracic duct in different groups were tested. The permeability, myeloperoxidase activity, and histopathological investigation of intestine and lung were evaluated. RESULTS: Cytokines and endotoxin levels in the portal vein were significantly increased in experimental groups compared with control group (P < 0.05), and that in the lymph from thoracic duct were significantly increased in PV O/R group compared with control and PV O/R + ML group (P < 0.05). Lung permeability and MPO activity in PV O/R group were significantly higher than those in control and PV O/R + ML group (P < 0.05); intestinal permeability in experimental groups were significantly higher with respect to control group. The lung injury score in PV O/R group was significantly higher than those in control and PV O/R + ML group (P < 0.05) and the intestinal injury scores in experimental groups were significantly higher than control group (P < 0.05). CONCLUSIONS: The gut-induced lung injury canine model made by PV O/R is successful, and mesenteric lymphatic ducts ligation decreases the degree of gut-induced lung injury in this model.

Hydrogen-rich water protects against inflammatory bowel disease in mice by inhibiting endoplasmic reticulum stress and promoting heme oxygenase-1 expression.

AIM: To investigate the therapeutic effect of hydrogen-rich water (HRW) on inflammatory bowel disease (IBD) and to explore the potential mechanisms involved. METHODS: Male mice were randomly divided into the following four groups: control group, in which the mice received equivalent volumes of normal saline (NS) intraperitoneally (ip); dextran sulfate sodium (DSS) group, in which the mice received NS ip (5 mL/kg body weight, twice per day at 8 am and 5 pm) for 7 consecutive days after IBD modeling; DSS + HRW group, in which the mice received HRW (in the same volume as the NS treatment) for 7 consecutive days after IBD modeling; and DSS + HRW + ZnPP group, in which the mice received HRW (in the same volume as the NS treatment) and ZnPP [a heme oxygenase-1 (HO-1) inhibitor, 25 mg/kg] for 7 consecutive days after IBD modeling. IBD was induced by feeding DSS to the mice, and blood and colon tissues were collected on the 7th d after IBD modeling to determine clinical symptoms, colonic inflammation and the potential mechanisms involved. RESULTS: The DSS + HRW group exhibited significantly attenuated weight loss and a lower extent of disease activity index compared with the DSS group on the 7th d (P < 0.05). HRW exerted protective effects against colon shortening and colonic wall thickening in contrast to the DSS group (P < 0.05). The histological study demonstrated milder inflammation in the DSS + HRW group, which was similar to normal inflammatory levels, and the macroscopic and microcosmic damage scores were lower in this group than in the DSS group (P < 0.05). The oxidative stress parameters, including MDA and MPO in the colon, were significantly decreased in the DSS + HRW group compared with the DSS group (P < 0.05). Simultaneously, the protective indicators, superoxide dismutase and glutathione, were markedly increased with the use of HRW. Inflammatory factors were assessed, and the results showed that the DSS + HRW group exhibited significantly reduced levels of TNF-α, IL-6 and IL-1ß compared with the DSS group (P < 0.05). In addition, the pivotal proteins involved in endoplasmic reticulum (ER) stress, including p-eIF2α, ATF4, XBP1s and CHOP, were dramatically reduced after HRW treatment in contrast to the control group (P < 0.05). Furthermore, HRW treatment markedly up-regulated HO-1 expression, and the use of ZnPP obviously reversed the protective role of HRW. In the DSS + HRW + ZnPP group, colon shortening and colonic wall thickening were significantly aggravated, and the macroscopic damage scores were similar to those of the DSS + HRW group (P < 0.05). The histological study also showed more serious colonic damage that was similar to the DSS group. CONCLUSION: HRW has a significant therapeutic potential in IBD by inhibiting inflammatory factors, oxidative stress and ER stress and by up-regulating HO-1 expression.