Hydrogen-rich Water Treatment Research Articles

The role of hydrogen-rich water in delaying the pulp breakdown of litchi fruit during postharvest storage

Previous studies have indicated that hydrogen-rich water (HW) treatment can delay fruit ripening and senescence. However, little is known about the HW-delaying pulp breakdown. In this study, eight physiological characteristics revealed that HW treatment delayed both pericarp browning and pulp breakdown of litchi fruit. To gain a comprehensive understanding of the changes in litchi pulp, a combination of multiple metabolomics and gene expression analyses was conducted, assessing 67 primary metabolites, 103 volatiles, 31 amino acids, and 13 crucial metabolite-related genes. Results showed that HW treatment promoted starch degradation, decelerated cell wall degradation and glycolysis, and maintained the flavor and quality of litchi fruit. Furthermore, HW treatment stimulated the production of volatile alcohols, aldehydes, ketones, olefins, and amino acids, which might play a vital role in HW-delaying pulp breakdown. This study sheds light on the mechanism by which HW delayed pulp breakdown by investigating small molecule metabolites and metabolic pathways.

Decoupling the mutual promotion of inflammation and oxidative stress mitigates cognitive decline and depression-like behavior in rmTBI mice by promoting myelin renewal and neuronal survival

BackgroundRepetitive mild traumatic brain injury (rmTBI) can lead to somatic, emotional, and cognitive symptoms that persist for years after the initial injury. Although the ability of various treatments to promote recovery after rmTBI has been explored, the optimal time window for early intervention after rmTBI is unclear. Previous research has shown that hydrogen-rich water (HRW) can diffuse through the blood-brain - barrier, attenuate local oxidative stress, and reduce neuronal apoptosis in patients with severe traumatic brain injury. However, research on the effect of HRW on rmTBI is scarce. AimsThe objectives of this study were to explore the following changes after rmTBI and HRW treatment: (i) temporal changes in inflammasome activation and oxidative stress-related protein expression through immunoblotting, (ii) temporal changes in neuron/myelin-related metabolite concentrations in vivo through magnetic resonance spectroscopy, (iii) myelin structural changes in late-stage rmTBI via immunofluorescence, and (iv) postinjury anxiety/depression-like behaviors and spatial learning and memory impairment. ResultsNLRP-3 expression in the rmTBI group was elevated at 7 and 14 DPI, and inflammasome marker levels returned to normal at 30 DPI. Oxidative stress persisted throughout the first month postinjury. HRW replacement significantly decreased Nrf2 expression in the prefrontal cortex and hippocampal CA2 region at 14 and 30 DPI, respectively. Edema and local gliosis in the hippocampus and restricted diffusion in the thalamus were observed on MR-ADC images. The tCho/tCr ratio in the rmTBI group was elevated, and the tNAA/tCr ratio was decreased at 30 DPI. Compared with the mice in the other groups, the mice in the rmTBI group spent more time exploring the open arms in the elevated plus maze (P < 0.05) and were more active in the maze (longer total distance traveled). In the sucrose preference test, the rmTBI group exhibited anhedonia. In the Morris water maze test, the latency to find the hidden platform in the rmTBI group was longer than that in the sham and HRW groups (P < 0.05). ConclusionEarly intervention with HRW can attenuate inflammasome assembly and reduce oxidative stress after rmTBI. These changes may restore local oligodendrocyte function, promote myelin repair, prevent axonal damage and neuronal apoptosis, and alleviate depression-like behavior and cognitive impairment.

Hydrogen-rich water: a key player in boosting wheat (Triticum aestivum L.) seedling growth and drought resilience

In the modern world, wheat, a vital global cereal and the second most consumed, is vulnerable to climate change impacts. These include erratic rainfall and extreme temperatures, endangering global food security. Research on hydrogen-rich water (HRW) has gained momentum in plant and agricultural sciences due to its diverse functions. This study examined the effects of different HRW treatment durations on wheat, revealing that the 4-h treatment had the highest germination rate, enhancing potential, vigor, and germination indexes. This treatment also boosted relative water content, root and shoot weight, and average lengths. Moreover, the 4-h HRW treatment resulted in the highest chlorophyll and soluble protein concentrations in seeds while reducing cell death. The 4-h and 5-h HRW treatments significantly increased H2O2 levels, with the highest NO detected in both root and shoot after 4-h HRW exposure. Additionally, HRW-treated seeds exhibited increased Zn and Fe concentrations, along with antioxidant enzyme activities (CAT, SOD, APX) in roots and shoots. These findings suggest that HRW treatment could enhance wheat seed germination, growth, and nutrient absorption, thereby increasing agricultural productivity. Molecular analysis indicated significant upregulation of the Dreb1 gene with a 4-h HRW treatment. Thus, it shows promise in addressing climate change effects on wheat production. Therefore, HRW treatment could be a hopeful strategy for enhancing wheat plant drought tolerance, requiring further investigation (field experiments) to validate its impact on plant growth and drought stress mitigation.

Hydrogen-rich water treatment improves fresh-cut Gastrodia elata quality by regulating reactive oxygen species metabolism and energy metabolism during low temperature storage

Hydrogen-rich water treatment improves fresh-cut Gastrodia elata quality by regulating reactive oxygen species metabolism and energy metabolism during low temperature storage

Hydrogen-rich water treatment targets RT1-Db1 and RT1-Bb to alleviate premature ovarian failure in rats.

Premature ovarian failure (POF) is defined as the cessation of ovarian function before the age of 40 years, imposing a significant health burden on patients. However, effective etiological therapy for POF is scarce. Thus, we aimed to explore the protective role and targets of hydrogen-rich water (HRW) in POF. Based on cyclophosphamide (CTX)-induced POF rat models, the protective role of HRW treatment was mainly determined through serum 17-β-estradiol (E2), follicle-stimulating hormone (FSH), anti-mullerian hormone (AMH) levels, ovarian histomorphological analysis, and TUNEL assay. Tandem mass tag (TMT)-based quantitative proteomic analysis was then conducted on ovarian tissues, and the targets of HRW in POF were identified integrating differential expression analysis, functional enrichment analysis, and interaction analysis. In HRW treatment of POF rats, the serum AMH and E2 levels significantly increased, and FSH level significantly reduced, indicating the protective role of HRW. After TMT quantitative proteomic analysis, a total of 16 candidate differentially expressed proteins (DEPs) were identified after the cross analysis of DEPs from POF vs. control and POF+HRW vs. POF groups, which were found to be significantly enriched in 296 GO terms and 36 KEGG pathways. The crucial targets, RT1-Db1 and RT1-Bb, were finally identified based on both protein-protein interaction network and GeneMANIA network. The HRW treatment could significantly alleviate the ovarian injury of POF rats; RT1-Db1 and RT1-Bb are identified as two crucial targets of HRW treatment in POF rats.

Hydrogen-rich water reduces cell damage by reducing excessive autophagy in mouse neuronal cells after oxygen glucose deprivation/reoxygenation

To investigate whether hydrogen-rich water exerts a protective effect against cellular injury by affecting the level of autophagy after oxygen glucose deprivation/reoxygenation (OGD/R) in a mouse hippocampal neuronal cell line (HT22 cells). HT22 cells in logarithmic growth phase were cultured in vitro. Cell viability was detected by cell counting kit-8 (CCK-8) assay to find the optimal concentration of Na2S2O4. HT22 cells were divided into control group (NC group), OGD/R group (sugar-free medium+10 mmol/L Na2S2O4 treated for 90 minutes and then changed to normal medium for 4 hours) and hydrogen-rich water treatment group (HW group, sugar-free medium+10 mmol/L Na2S2O4 treated for 90 minutes and then changed to medium containing hydrogen-rich water for 4 hours). The morphology of HT22 cells was observed by inverted microscopy; cell activity was detected by CCK-8 method; cell ultrastructure was observed by transmission electron microscopy; the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin-1 was detected by immunofluorescence; the protein expression of LC3II/I and Beclin-1, markers of cellular autophagy, was detected by Western blotting. Inverted microscopy showed that compared with the NC group, the OGD/R group had poor cell status, swollen cytosol, visible cell lysis fragments and significantly lower cell activity [(49.1±2.7)% vs. (100.0±9.7)%, P < 0.01]; compared with the OGD/R group, the HW group had improved cell status and remarkably higher cell activity [(63.3±1.8)% vs. (49.1±2.7)%, P < 0.01]. Transmission electron microscopy showed that the neuronal nuclear membrane of cells in the OGD/R group was lysed and a higher number of autophagic lysosomes were visible compared with the NC group; compared with the OGD/R group, the neuronal damage of cells in the HW group was reduced and the number of autophagic lysosomes was notably decreased. The results of immunofluorescence assay showed that the expressions of LC3 and Beclin-1 were outstandingly enhanced in the OGD/R group compared with the NC group, and the expressions of LC3 and Beclin-1 were markedly weakened in the HW group compared with the OGD/R group. Western blotting assay showed that the expressions were prominently higher in both LC3II/I and Beclin-1 in the OGD/R group compared with the NC group (LC3II/I: 1.44±0.05 vs. 0.37±0.03, Beclin-1/β-actin: 1.00±0.02 vs. 0.64±0.01, both P < 0.01); compared with the OGD/R group, the protein expression of both LC3II/I and Beclin-1 in the HW group cells were notably lower (LC3II/I: 0.54±0.02 vs. 1.44±0.05, Beclin-1/β-actin: 0.83±0.07 vs. 1.00±0.02, both P < 0.01). Hydrogen-rich water has a significant protective effect on OGD/R-causing HT22 cell injury, and the mechanism may be related to the inhibition of autophagy.

The Role and Mechanism of Hydrogen-Rich Water in the Cucumis sativus Response to Chilling Stress

Cucumber is a warm climate vegetable that is sensitive to chilling reactions. Chilling can occur at any period of cucumber growth and development and seriously affects the yield and quality of cucumber. Hydrogen (H2) is a type of antioxidant that plays a critical role in plant development and the response to stress. Hydrogen-rich water (HRW) is the main way to use exogenous hydrogen. This study explored the role and mechanism of HRW in the cucumber defense response to chilling stress. The research results showed that applying 50% saturated HRW to the roots of cucumber seedlings relieved the damage caused by chilling stress. The growth and development indicators, such as plant height, stem diameter, leaf area, dry weight, fresh weight, and root length, increased under the HRW treatment. Photosynthetic efficiency, chlorophyll content, and Fv/Fm also improved and reduced energy dissipation. In addition, after HRW treatment, the REC and MDA content were decreased, and membrane lipid damage was reduced. NBT and DAB staining results showed that the color was lighter, and the area was smaller under HRW treatment. Additionally, the contents of O2- and H2O2 also decreased. Under chilling stress, the application of HRW increased the activity of the antioxidases SOD, CAT, POD, GR, and APX and improved the expression of the SOD, CAT, POD, GR, and APX antioxidase genes. The GSSG content was reduced, and the GSH content was increased. In addition, the ASA content also increased. Therefore, exogenous HRW is an effective measure for cucumber to respond to chilling stress.

Hydrogen-rich water treatment increased several phytohormones and prolonged the shelf life in postharvest okras.

Hydrogen-rich water (HRW) treatment has been reported to delay the softening and senescence of postharvest okras, but its regulatory mechanism remains unclear. In this paper, we investigated the effects of HRW treatment on the metabolism of several phytohormones in postharvest okras, which act as regulatory molecules in fruit ripening and senescence processes. The results showed that HRW treatment delayed okra senescence and maintained fruit quality during storage. The treatment upregulated all of the melatonin biosynthetic genes such as AeTDC, AeSNAT, AeCOMT and AeT5H, contributing to the higher melatonin content in the treated okras. Meanwhile, increased transcripts of anabolic genes but lower expression of catabolic genes involved in indoleacetic acid (IAA) and gibberellin (GA) metabolism were observed in okras when treated with HRW, which was related to the enhanced levels of IAA and GA. However, the treated okras experienced lower abscisic acid (ABA) content as compared to the non-treated fruit due to the down-regulation of its biosynthetic genes and up-regulation of the degradative gene AeCYP707A. Additionally, there was no difference in γ-aminobutyric acid between the non-treated and HRW-treated okras. Collectively, our results indicated that HRW treatment increased levels of melatonin, GA and IAA, but decreased ABA content, which ultimately delayed fruit senescence and prolonged shelf life in postharvest okras.

Molecular Hydrogen Prevents Osteoclast Activation in a Glucocorticoid-Induced Osteoporosis Zebrafish Scale Model.

Antioxidants represent a powerful tool for many human diseases and, in particular, molecular hydrogen has unique characteristics that make it a very promising therapeutic agent against osteoporosis. Zebrafish scales offer an innovative model in which new therapeutic approaches against secondary osteoporosis are tested. Scale bone loss obtained by prednisolone (PN) treatment is characterized by increased osteoclast activity and decreased osteoblast activity highlighted with bone enzymatic assays. We used this read-out system to test the therapeutic effects of hydrogen-rich water (HRW), an innovative antioxidant approach. HRW prevented osteoclast activation and bone loss in PN-treated fish scales, as verified by both biochemical and histochemical tartrate-resistant alkaline phosphatase assays. On the other hand, HRW treatment did not prevent PN-dependent osteoblast suppression, as measured by alkaline phosphatase activity. Moreover, HRW treatment did not facilitate the reparation of resorption lacunae induced in scales by PN. Our study highlighted a specific effect of HRW on adult osteoclast activity but not in osteoblasts, introducing an intriguing new antioxidant preventive approach against osteoporosis.

Hydrogen-Rich Water Treatment of Fresh-Cut Kiwifruit with Slightly Acidic Electrolytic Water: Influence on Antioxidant Metabolism and Cell Wall Stability.

The synergistic impact of hydrogen-rich water (HRW, 394 ppb) and slightly acidic electrolyzed water (SAEW, pH of 6.25 ± 0.19) on the antioxidant metabolism of fresh-cut kiwifruit during storage was investigated (temperature: (3 ± 1) °C, humidity: 80%-85%). Compared with control group, H+S treatment increased the contents of active oxygen-scavenging enzymes (SOD, CAT, POD, and APX) and inhibited the increase of O2•- and H2O2 contents during the storage of fresh-cut kiwifruit. Meanwhile, H+S treatment could reduce the activities of the cell wall-degrading enzymes PG, PME, PL, Cx, and β-Gal, inhibit the formation of soluble pectin, delay the degradation rate of propectin, cellulose, and pseudocellulose, and maintain higher fruit hardness and chewability. The results showed that H+S treatment could enhance free radical scavenging ability and reduce the cell wall metabolism of fresh-cut kiwifruit, maintaining the good texture found in fresh-cut fruit.

Evaluation of the impact of hydrogen-rich water on the quality attribute notes of butter.

The effects of washing raw butter with hydrogen-rich water (HRW), prepared with hydrogen (H2) and/or magnesium (Mg), on butter quality were investigated in this research paper. During the washing process, titratable acidity (TA) decreased by 12% for all washed samples. During the storage period, TA increased by 28% and 93% (control), 14% and 58% (H2), and 10% and 66% (Mg) for the 60th and 90th days, respectively. Peroxide value (mEq O2/kg) increased to 2.76 and 8.83 (control), 1.92 and 7.25 (H2), and 2.02 and 8.12 (Mg) for the 60th and 90th days. HRW samples showed the lowest acid degree value (ADV) and the highest color notes (L*, C*, and h). The HRW treatment of raw butter has shown improving effects on the product without any harmful residuals in the final product or the environment.

Effects of hydrogen‐rich water on the nutritional properties, volatile profile and texture of germinated brown rice

SummaryIn the present study, we investigated the effects of hydrogen‐rich water (HRW) on the germination efficiency, chemical composition, biofunctional and volatile compounds, and texture of germinated brown rice (GBR). Our results showed that HRW (1.5 mg L−1) treatment significantly increased the germination efficiency of GBR by changing the migration of water during soaking. A chemical composition analysis showed that HRW treatment changed the contents of some biofunctional compounds and volatile components; increased the contents of soluble dietary fibre, thiamine, niacin, γ‐aminobutyric acid, some amino acids and K, P, Mn and Fe; and decreased the contents of starch and Na of GBR compared with those of GBR treated with regular pure water. Moreover, HRW treatment also improved the texture of GBR due to the changes in the ultrastructure of the bran layer and the decrease in insoluble dietary fibre. In summary, HRW is suggested to be applied in the processing of GBR since it can increase germination efficiency and improve the nutritional and eating qualities of GBR.

Hydrogen-rich water maintains the quality of Rosa roxburghii by regulating AsA biosynthesis and regeneration

Rosa roxburghii is of great interest due to its high ascorbic acid (AsA) content, however AsA is readily lost after harvest. This study aimed to explore the effect of hydrogen-rich water (HRW) treatment on AsA metabolism in postharvest R. roxburghii. Results showed that HRW treatment maintained fruit quality and higher AsA content of R. roxburghii during the assay period. HRW treatment up-regulated the expression of AsA synthesis-related genes such as RrGMP, RrGME, RrGGP, RrGalDH, RrGalLDH and RrGalUR, but barely regulated the expression of RrGPP and RrMIOX. HRW promoted the activity and gene expression of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) in the AsA-GSH cycle. More interestingly, seven RrMYBs were significantly upregulated by HRW treatment and their expression trends were highly correlated with AsA content, suggesting that RrMYBs might transcriptionally regulate AsA synthesis and regeneration in postharvest. Our results emphasized that HRW reduced AsA loss, and positively regulated the expression of numerous AsA synthesis and regeneration genes and RrMYBs, thus illustrating a close link between RrMYBs and AsA synthesis and regeneration in postharvest R. roxburghii fruit.

Hydrogen-Rich Water as a Novel Therapeutic Strategy for the Affective Disorders Linked with Chronic Neuropathic Pain in Mice.

Neuropathic pain manifested with allodynia and hyperalgesia usually becomes a chronic condition accompanied with mood disorders. Clinical therapies for neuropathic pain are still unsatisfactory with notable side effects. Recent studies have reported the protective role of molecular hydrogen (H2) in different diseases including neurological disorders, such as Alzheimer’s as well as its antidepressant activities in animals with chronic stress. This study explored the effects of treatment with hydrogen-rich water (HRW) in male mice with neuropathic pain induced by the chronic constriction of the sciatic nerve (CCI) and the accompanying affective deficits. The likely pathways implied in the HRW analgesic activity, as well as the interaction between heme oxygenase 1 (HO-1) enzyme and H2 during neuropathic pain were also studied. The results showed: (i) the inhibitory effects of the repetitive treatment with HRW on the allodynia and hyperalgesia provoked by CCI; (ii) the anxiolytic and antidepressant actions of HRW in animals with neuropathic pain; (iii) the contribution of the antioxidant enzymes (HO-1 and NAD(P)H: quinone oxidoreductase 1) and the ATP sensitive potassium channels in the painkiller activities of HRW during neuropathic pain; (iv) a positive interaction between the HO-1 and H2 systems in inhibiting the CCI-induced neuropathy; and (v) the antioxidant, antinociceptive, anti-inflammatory and/or antiapoptotic features of HRW treatment in the dorsal root ganglia and/or amygdala of sciatic nerve-injured mice. This study demonstrates new protective actions of H2 and suggests that treatment with HRW might be an interesting therapeutic strategy for chronic neuropathic pain and its associated mood disorders.

Hydrogen-rich water delays fruit softening and prolongs shelf life of postharvest okras

The effect of hydrogen-rich water (HRW) treatment on softening, cell wall components and cell wall metabolic genes in okras after harvest was studied. The results showed that HRW treatment could maintain fruit firmness and delay softening, thereby prolonging shelf life in okras during storage. The treated okras displayed significantly lower levels water- and chelate-soluble pectins while higher contents of Na2CO3-soluble pectin, hemicellulose and cellulose. The cell wall biosynthesis was maintained by HRW treatment via up-regulating genes involved in biosynthesis of pectin, hemicellulose and cellulose at the beginning of storage. On the contrary, the treatment could inhibit the cell wall disassembly due to the down-regulation of numerous cell wall degradative genes including AePME, AeGAL and AeCX at the end of storage. Taken together, our results suggested that HRW treatment delayed softening and extended shelf life in postharvest okras through modifying cell wall biosynthesis and disassembly at different times of storage.

Transcriptome and Metabonomics Combined Analysis Revealed the Defense Mechanism Involved in Hydrogen-Rich Water-Regulated Cold Stress Response of Tetrastigma hemsleyanum.

The poor resistance to cold stress conditions has become the bottleneck problem in Tetrastigma hemsleyanum (T. hemsleyanum) planting industry. Exogenous hydrogen (H2) plays an important role in improving stress resistance in plants. However, the key factors and regulatory network of plants in response to hydrogen-rich water (HRW) treatment under environmental stress are not clear. Here, we conducted integrative analyses of metabolome and transcriptome profiles to reveal the defense mechanism involved in the HRW-regulated cold stress response of T. hemsleyanum. The application of 75% HRW could alleviate stress damage by decreasing stomatal apparatus density and significantly increasing photosynthetic efficiency and mitigating physiological indexes of resistance, such as Pn, Cond, MDA, SOD, etc., which were changed by cold stress conditions. A total of 7,883 DEGs and 439 DEMs were identified. DEGs were the most relevant to phenylpropanoid, isoflavonoid, monoterpenoid, and flavonoid biosynthesis pathways. Using gene co-expression analysis (WGCNA), we identified one gene module that showed a strong correlation between total antioxidant capacity and transpiration rate. Trend analysis indicated that the phenylpropanoid biosynthesis pathway played a major role in the transcription and metabolism process of HRW treatment under cold stress. Based on the integrated analysis of genes and metabolites, the results showed cold stress upregulated the expression of PAL, CHS, COMT, CCR, AtBG1, etc., resulting in the accumulation of coniferyl alcohol and eriodictyol contents in T. hemsleyanum under cold stress, but the 75% HRW treatment could attenuate the enhancement. The study not only identified the main strategy of HRW protection against cold stress but also provided candidate genes for flavonoid biosynthesis, so as to better improve cold tolerance through molecular breeding techniques.

Integrated Metabolomic and Transcriptomic Analyses to Understand the Effects of Hydrogen Water on the Roots of Ficus hirta Vahl.

Wuzhimaotao (Ficus hirta Vahl) is an important medicinal and edible plant in China. The extract from the roots of Ficus hirta Vahl contains phenylpropanoid compounds, such as coumarins and flavonoids, which are the main active components of this Chinese herbal medicine. In this study, we analyzed the transcriptomic and metabolomic data of the hydrogen-water-treated roots of Ficus hirta Vahl and a control group. The results showed that many genes and metabolites were regulated in the roots of Ficus hirta Vahl that were treated with hydrogen water. Compared with the control group, 173 genes were downregulated and 138 genes were upregulated in the hydrogen-rich water treatment group. Differential metabolite analysis through LC-MS showed that 168 and 109 metabolites had significant differences in positive and negative ion mode, respectively. In the upregulated metabolites, the main active components of Wuzhimaotao, such as the phenylpropane compounds naringin, bergaptol, hesperidin, and benzofuran, were found. Integrated transcriptomic and metabolomic data analysis showed that four and one of the most relevant pathways were over enriched in positive and negative ion mode, respectively. In the relationship between metabolites and DEGs, phenylpropanoid biosynthesis and metabolism play an important role. This indicates that phenylpropanoid biosynthesis and metabolism may be the main metabolic pathways regulated by hydrogen water. Our transcriptome analysis showed that most of the DEGs with |log2FC| ≥ 1 are transcription factor genes, and most of them are related to plant hormone signal transduction, stress resistance, and secondary metabolism, mainly phenylpropanoid biosynthesis and metabolism. This study provides important evidence and clues for revealing the botanical effect mechanism of hydrogen and a theoretical basis for the application of hydrogen agriculture in the cultivation of Chinese herbal medicine.

The Therapeutic Effects of Oral Intake of Hydrogen Rich Water on Cutaneous Wound Healing in Dogs.

This study explored the effects of drinking Hydrogen-rich water (HRW) on skin wound healing in dogs. Eight circular wounds were analyzed in each dog. The experimental group was treated with HRW thrice daily, while the control group was provided with distilled water (DW). The wound tissues of dogs were examined histopathologically. The fibroblasts, inflammatory cell infiltration, the average number of new blood vessels, and the level of malondialdehyde (MDA) and superoxide dismutase (SOD) activity in the skin homogenate of the wound was measured using the corresponding kits. The expressions of Nrf-2, HO-1, NQO-1, VEGF, and PDGF were measured using the real-time fluorescence quantitative method. We observed that HRW wounds showed an increased rate of wound healing, and a faster average healing time compared with DW. Histopathology showed that in the HRW group, the average thickness of the epidermis was significantly lower than the DW group. The average number of blood vessels in the HRW group was higher than the DW group. The MDA levels were higher in the DW group than in the HRW group, but the SOD levels were higher in the HRW group than in the DW group. The results of qRT-PCR showed that the expression of each gene was significantly different between the two groups. HRW treatment promoted skin wound healing in dogs, accelerated wound epithelization, reduced inflammatory reaction, stimulated the expression of cytokines related to wound healing, and shortened wound healing time.

Effect of hydrogen-rich water on the lactic acid level in metformin-treated diabetic rats under hypoxia.

The present study aims to investigate the impact of hydrogen-rich water on the lactic acid level in metformin-treated diabetic rats under hypoxia. Thirty Sprague-Dawley rats were randomly divided into five groups, including normal diet group, and diabetes model (DM) group, DM + metformin treatment (DMM) group, DMM + hypoxia treatment (DMMH) group and DMMH + hydrogen-rich water (DMMHR) group. We found that the levels of lactic acid, pyruvate and lactate dehydrogenase were significantly lower in the blood of DMMHR group than DMMH group. Superoxide dismutase and glutathione levels in liver and heart were significantly higher in DMMH group after hydrogen-rich water treatment, while malondialdehyde and oxidized glutathione levels were decreased in DMMHR group when compared with DMMH group, which indicates that hydrogen-rich water could reduce oxidative stress. qPCR analysis demonstrated that that pro-apoptotic genes Bax/Caspase-3 were upregulated in DM group and metformin treatment suppressed their upregulation (DMM group). However, hypoxic condition reversed the effect of metformin on apoptotic gene expression, and hydrogen-rich water showed little effect on these genes under hypoxia. HE staining showed that hydrogen-rich water prevented myocardial fiber damages under hypoxia. In summary, we conclude that hydrogen-rich water could prevent lactate accumulation and reduce oxidant stress in diabetic rat model to prevent hypoxia-induced damages. It could be served as a potential agent for diabetes patients with metformin treatment to prevent lactic acidosis and reduce myocardial damages under hypoxic conditions.

Hydrogen-rich water promotes the formation of bulblets in Lilium davidii var. unicolor through regulating sucrose and starch metabolism.

Main conclusion HRW increased the content of starch and sucrose via regulating a series of sucrose and starch synthesis genes, which induced the formation of bulblets and adventitious roots of Lilium davidii var. unicolor. Hydrogen gas (H2), as a signaling molecule, has been reported to be involved in plant growth and development. Here, the effect of hydrogen-rich water (HRW) on the formation of bulblets and adventitious roots in the scale cuttings of Lilium davidii var. unicolor and its mechanisms at the molecular levels were investigated. The results revealed that compared with distilled water treatment (Con), the number of bulblets and adventitious roots were significantly promoted by different concentrations of HRW treatment. Treatment with 100% HRW obtained the most positive effects. RNA sequencing (RNA-seq) analysis found that compared with Con, a total of 1702 differentially expressed genes (DEGs, upregulated 552 DEGs, downregulated 1150 DEGs) were obtained under HRW treatment. The sucrose and starch metabolism, cysteine and methionine metabolism and phenylalanine metabolism were significantly enriched in the analysis of the Kyoto encyclopedia of genes and genomes (KEGG). In addition, the genes involved in carbohydrate metabolism were significantly upregulated or downregulated (upregulated 22 DEGs, downregulated 15 DEGs), indicating that starch and sucrose metabolism held a central position. The expressions of 12 DEGs were identified as coding for key enzymes in metabolism of carbohydrates was validated by qPCR during bulblet formation progress. RNA-seq analysis and expression profiles indicated that the unigene levels such as glgc, Susy, otsA and glgP, BMY and TPS were well correlated with sucrose and starch metabolism during HRW-induced bulblet formation. The change of key enzyme content in starch and sucrose metabolism pathway was explored during bulblet formation in Lilium under HRW treatment. Meanwhile, compared with Con, 100% HRW treatment increased the levels of sucrose and starch, and decreased the trehalose content, which were agreed with the expression pattern of DEGs related to the biosynthesis pathway of sucrose, starch and trehalose. Therefore, this study suggested that HRW could promote the accumulation of sucrose and starch contents in mother scales, and decreased the trehalose content, this might provide more energy for bulblet formation.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00425-021-03762-6.