Stem Cell Res Ther. 2019 May 21;10(1):145. doi: 10.1186/s13287-019-1241-x.
Molecular hydrogen suppresses glioblastoma growth via inducing the glioma stem-like cell differentiation.
Glioblastoma (GBM) is the most common type of primary malignant brain tumor. Molecular hydrogen has been considered a preventive and therapeutic medical gas in many diseases including cancer. In our study, we sought to assess the potential role of molecular hydrogen on GBM.
The in vivo studies were performed using a rat orthotopic glioma model and a mouse subcutaneous xenograft model. Animals inhaled hydrogen gas (67%) 1 h two times per day. MR imaging studies were performed to determine the tumor volume. Immunohistochemistry (IHC), immunofluorescence staining, and flow cytometry analysis were conducted to determine the expression of surface markers. Sphere formation assay was performed to assess the cancer stem cell self-renewal capacity. Assays for cell migration, invasion, and colony formation were conducted.
The in vivo study showed that hydrogen inhalation could effectively suppress GBM tumor growth and prolong the survival of mice with GBM. IHC and immunofluorescence staining demonstrated that hydrogen treatment markedly downregulated the expression of markers involved in stemness (CD133, Nestin), proliferation (ki67), and angiogenesis (CD34) and also upregulated GFAP expression, a marker of differentiation. Similar results were obtained in the in vitro studies. The sphere-forming ability of glioma cells was also suppressed by hydrogen treatment. Moreover, hydrogen treatment also suppressed the migration, invasion, and colony-forming ability of glioma cells.
* Together, these results indicated that molecular hydrogen may serve as a potential anti-tumor agent in the treatment of GBM.
Oncol Rep. 2019 Jan;41(1):301-311. doi: 10.3892/or.2018.6841. Epub 2018 Nov 1.
Hydrogen gas restores exhausted CD8+ T cells in patients with advanced colorectal cancer to improve prognosis.
Exhausted cluster of differentiation (CD)8+ T cells lose immunological activity due to mitochondrial dysfunction caused by peroxisome proliferator‑activated receptor γ coactivator 1α (PGC‑1α) inactivation, resulting in a poor prognosis in patients with cancer. As hydrogen gas was recently reported to activate PGC‑1α, the present study investigated whether it restores exhausted CD8+ T cells to improve prognosis in patients with stage IV colorectal cancer.
* A total of 55 patients with histologically and clinically diagnosed stage IV colorectal carcinoma were enrolled between July 2014 and July 2017.
The patients inhaled hydrogen gas for 3 h/day at their own homes and received chemotherapy at the Tamana Regional Health Medical Center (Tamana, Kumamoto, Japan). The CD8+ T cells were isolated from the peripheral blood and their phenotype was analyzed by flow cytometry. It was found that exhausted terminal programmed cell death 1 (PD‑1)+ CD8+ T cells in the peripheral blood are independently associated with worse progression‑free survival (PFS) and overall survival (OS). Notably, hydrogen gas decreased the abundance of exhausted terminal PD‑1+ CD8+ T cells, increased that of active terminal PD‑1‑ CD8+ T cells, and improved PFS and OS times, suggesting that the balance between terminal PD1+ and PD1‑ CD8+ T cells is critical for cancer prognosis.
Therefore, a novel system for patient classification (category 1‑4) was developed in the present study based on these two indices to assist in predicting the prognosis and therapeutic response. Collectively, the present results suggested that hydrogen gas reverses imbalances toward PD‑1+ CD8+ T cells to provide an improved prognosis.
Med Gas Res. 2019 Jan 9;8(4):144-149. doi: 10.4103/2045-9912.248264. eCollection 2018 Oct-Dec.
Inhalation of hydrogen gas elevates urinary 8-hydroxy-2'-deoxyguanine in Parkinson's disease.
Hyposmia is one of the earliest and the most common symptoms in Parkinson's disease (PD). The benefits of hydrogen water on motor deficits have been reported in animal PD models and PD patients, but the effects of hydrogen gas on PD patients have not been studied.
* We evaluated the effect of inhalation of hydrogen gas on olfactory function, non-motor symptoms, activities of daily living, and urinary 8-hydroxy-2'-deoxyguanine (8-OHdG) levels by a randomized, double-blinded, placebo-controlled, crossover trial with an 8-week washout period in 20 patients with PD.
Patients inhaled either ~1.2-1.4% hydrogen-air mixture or placebo for 10 minutes twice a day for 4 weeks.
** Inhalation of low dose hydrogen did not significantly influence the PD clinical parameters, but it did increase urinary 8-OHdG levels by 16%.
This increase in 8-OHdG is markedly less than the over 300% increase in diabetes, and is more comparable to the increase after a bout of strenuous exercise. Although increased reactive oxygen species is often associated with toxicity and disease, they also play essential roles in mediating cytoprotective cellular adaptations in a process known as hormesis. Increases of oxidative stress by hydrogen have been previously reported, along with its ability to activate the Nrf2, NF-κB pathways, and heat shock responses. Although we did not observe any beneficial effect of hydrogen in our short trial, we propose that the increased 8-OHdG and other reported stress responses from hydrogen may indicate that its beneficial effects are partly or largely mediated by hormetic mechanisms.
PLoS One. 2018 Dec 26;13(12):e0208313. doi: 10.1371/journal.pone.0208313. eCollection 2018.
Quantification of hydrogen production by intestinal bacteria that are specifically dysregulated in Parkinson's disease.
Oral administration of hydrogen water ameliorates Parkinson's disease (PD) in rats, mice, and humans. We previously reported that the number of putative hydrogen-producing bacteria in intestinal microbiota is low in PD compared to controls. We also reported that the amount of hydrogen produced by ingestion of lactulose is low in PD patients. The decreased hydrogen production by intestinal microbiota may be associated with the development and progression of PD. We measured the amount of hydrogen production using gas chromatography by seven bacterial strains, which represented seven major intestinal bacterial groups/genera/species. Blautia coccoides and Clostridium leptum produced the largest amount of hydrogen. Escherichia coli and Bacteroides fragilis constituted the second group that produced hydrogen 34- to 93-fold lower than B. coccoides. Bifidobacterium pseudocatenulatum and Atopobium parvulum constituted the third group that produced hydrogen 559- to 2164-fold lower than B. coccoides. Lactobacillus casei produced no detectable hydrogen. Assuming that taxonomically neighboring strains have similar hydrogen production, we simulated hydrogen production using intestinal microbiota that we previously reported, and found that PD patients produce a 2.2-fold lower amount of intestinal hydrogen compared to controls. The lower amount of intestinal hydrogen production in PD was also simulated in cohorts of two other countries. The number of hydrogen-producing intestinal bacteria may be associated with the development and progression of PD. Further studies are required to prove its beneficial effect.
Int Immunopharmacol. 2018 Dec;65:366-372. doi: 10.1016/j.intimp.2018.10.012. Epub 2018 Oct 26.
Protective effects of hydrogen gas against sepsis-induced acute lung injury via regulation of mitochondrial function and dynamics.
Lungs are one of the most common target organs of sepsis . Hydrogen gas (H2), which has selective anti-oxidative effects, can be effectively used to treat septic mice. Mitochondrial dysfunction and dynamics play important roles in sepsis-induced organ damage.
By using cecal ligation and puncture (CLP), a classic septic model, we explored the role of 2% H2 treatment in sepsis-induced acute lung injury (ALI) linked to mitochondrial function and dynamics. We randomized male Institute for Cancer Research (ICR) mice into 4 groups: sham, sham + H2, CLP and CLP + H2. At 24 h after CLP or sham operations, we used histological examination and transmission electron microscopy (TEM) to observe lung slices. We analyzed oxygenation index (PaO2/FiO2), mitochondrial-membrane potential (MMP), adenosine triphosphate (ATP) levels, respiration control ratio (RCR) and mitochondrial-respiration complex activities (I and II) using commercial kits, and dynamin-related protein 1 (Drp1) and mitofusin-2 (MFN2) using Western blot.
Therapy with 2% H2 increased PaO2/FiO2 ratios, MMP and ATP levels, RCR, complex I activity and MFN2 expression but decreased histological score and Drp1 levels in the presence of sepsis.
* These data indicated that inhalation of 2% H2 to regulate mitochondrial function and dynamics may be a promising therapeutic strategy for lung injuries induced by severe sepsis.
Gan To Kagaku Ryoho. 2018 Oct;45(10):1475-1478.
[Immunological Effect of Hydrogen Gas-Hydrogen Gas Improves Clinical Outcomes of Cancer Patients].
[Article in Japanese]
It has been reported that PD-1-expressing CD8+ T cells in the peripheral blood of cancer patients are associated with poor cancer prognosis. In addition, these cells are in a state of energy shortage caused by mitochondrial dysfunction with a low level of PGC-1a. Recently, hydrogen gas was reported to activate PGC-1a, leading to the enhancement of mitochondrial activity. In the present study, we investigated whether hydrogen gas influences the proportion of PD-1+ CD8+ T cells in the peripheral blood of 55 Stage IV colorectal carcinoma patients. We found that the proportion of terminal PD-1+ CD8+ T cells was an independent factor for poor prognosis. We also found that the proportion of terminal PD-1+ CD8+ T cells was reduced in 35 out of 55 patients(63.6%)and was increased in 39 out of 55 patients(70.9%)after treatment with hydrogen gas. The ratio of the terminal PD-1+ CD8+ T cells after hydrogen gas treatment to that before hydrogen gas treatment(terminal PD-1+ CD8+ T cell ratio)was found to be an independent factor predicting PFS and OS. Out of another 26 patients treated with nivolumab, 14 patients treated with a combined therapy of hydrogen gas and nivolumab showed a significantly longer OS than the remaining 12 patients who were treated with nivolumab alone. These results suggest that hydrogen gas improves the prognosis of cancer patients by reducing the proportion of terminal PD-1+ CD8+ T cells.
Methods Mol Biol. 2019;1881:129-151. doi: 10.1007/978-1-4939-8876-1_11.
Measurement of Leukemic B-Cell Growth Kinetics in Patients with Chronic Lymphocytic Leukemia.
Cell proliferation plays a central role in the pathogenesis of every neoplastic disease as well as many other types of illness. Labeling of newly replicated DNA with deuterium (2H), a nonradioactive isotope of hydrogen, administered to the patients in drinking water (2H2O) is a safe and reliable method to measure the in vivo birth rates of cells. Here, we describe a protocol to measure chronic lymphocytic leukemia B-cell birth/proliferation and death rates over time using this approach.