NaHS Treatment Research Articles

Nitric oxide is involved in hydrogen sulfide-induced adventitious rooting in tomato (Solanum lycopersicum).

Nitric oxide (NO) and hydrogen sulfide (H2 S) are signalling molecules that regulate adventitious rooting in plants. However, little is known about the cross-talk between NO and H2 S during adventitious rooting. Tomato (Solanum lycopersicum L.) explants were used to investigate the roles of and relationships between NO and H2 S during rooting. Effects of the NO donor sodium nitroprusside (SNP) and the H2 S donor sodium hydrosulfide (NaHS) on adventitious rooting were dose-dependent, and the greatest biological responses were observed under 25μM SNP and 50μM NaHS. The positive effect of NaHS was reversed by the NO scavenger 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), indicating that the H2 S-induced response was partially NO-dependent. Peroxidase (POD), polyphenol oxidase (PPO), and superoxide dismutase (SOD) activities significantly increased by SNP and NaHS treatment, and indoleacetic acid oxidase (IAAO) activity and the O2 - and H2 O2 content significantly decreased by SNP and NaHS treatment. SNP and NaHS treatment also increased the content of soluble sugar and protein and indole-3-acetic acid (IAA). cPTIO significantly mitigated the increases in POD, PPO and SOD activity and soluble sugar, protein and IAA content induced by NaHS. SNP and NaHS upregulated the expression of auxin-related genes (ARF4 and ARF16 ), cell cycle-related genes (CYCD3 , CYCA3 and CDKA1 ), and antioxidant-related genes (TPX2 , SOD and POD ); whereas cPTIO significantly inhibited the increase in the expression of these genes induced by NaHS. Overall, these results show that NO may be involved in H2 S-induced adventitious rooting by regulating the activity of rooting-related enzymes, the expression of related genes, and the content of various nutrients.

Exogenous H2S Protects against Septic Cardiomyopathy by Inhibiting Autophagy through the AMPK/mTOR Pathway

Background Given the cardioprotective role of autophagy, this study aimed to investigate the protective effect of exogenous H2S (NaHS) on infectious cardiomyopathy through the inhibition of the AMPK/mTOR pathway. Methods In this study, sepsis models were established by cecal ligation and puncture (CLP) induction in vivo and intraperitoneal injection of NaHS was performed. Autophagy- and apoptosis-related proteins were observed by western blot, isolated myocardial tissue morphology was observed by hematoxylin-eosin (H&E) staining, and myocardial apoptosis was evaluated by the tunnel method. The ultrastructure of autophagy was observed by using an electron transmission electron microscope. Results In an SD rat model of cecum ligation puncture-induced sepsis, the level of autophagy-related proteins was significantly increased, and hematoxylin and eosin staining showed irregular myocardial bands and swollen cardiomyocytes. Following NaHS treatment, the level of autophagy-related proteins decreased, and electron transmission microscopy revealed decreased autophagosomes. Echocardiography suggested an increase in ejection fraction and significant relief of myocardial inhibition. Conclusions Our results suggest that NaHS treatment can attenuate the cellular damage caused by excessive autophagy through the AMPK/mTOR pathway.

Combined application of H2S and a plant growth promoting strain JIL321 regulates photosynthetic efficacy, soil enzyme activity and growth-promotion in rice under salt stress

Salinity stress is one of the most harmful abiotic stresses that inhibit crop growth and grain yield. In this study, a salt-tolerant bacterium was isolated from the soil of the rice rhizosphere and named Myroides sp. JIL321, based on the results of the phylogenetic tree analysis. The strain JIL321 tolerated up to 1, 283.37 mM of NaCl and exhibited positive plant growth-promoting traits, such as the production of indole acetic acid (IAA) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Therefore, the effects of JIL321 on rice (Oryza sativa L.) under salinity stress were determined. The inoculation of strain JIL321 significantly increased the chlorophyll content and the accumulation of osmotic adjustment substances, such as proline and soluble sugars, in rice expose to salt stress. Additionally, strain JIL321 inoculation significantly enhanced the activities of some enzymes commonly found in soil, such as urease, invertase and catalase. Moreover, the production of hydrogen sulfide (H2S), a pivotal signaling molecule, was also induced in rice by salt stress. Treatment with sodium hydrogen sulfide (NaHS, H2S donor) improved salt stress tolerance of the rice, while treatment with hypotaurine (HT, H2S scavenger) significantly suppressed it. Interestingly, NaHS treatment also improved the production of IAA and ACC deaminase in strain JIL321 under 0 mM and 150 mM salt concentrations. The combined treatment of JIL321 and NaHS could further improve the growth of salt-stressed rice seedlings, most likely due to the interaction effect between H2S and strain JIL321. To our knowledge, this study is the first to demonstrate that the combined use of H2S and plant growth-promoting bacteria could alleviate the adverse effects of salt stress on rice plants, and further verifies the novel role of H2S as a signaling molecule that enhance the tolerance of plant to abiotic stresses.

Protective effect of hydrogen sulfide against stress-induced lung injury: involvement of Nrf2, NFκB/iNOS, and HIF-1α signaling pathways

Stress is a common phenomenon that is attracting increasing attention. Hydrogen sulfide (H2S) is a gasotransmitter that plays an important role in many physiological and pathological events. Our study aimed to estimate the effect and the underlying mechanisms of the H2S donor, sodium hydrosulfide (NaHS), against immobilization stress (IS)-induced lung injury. Forty adult male rats were classified into control group, NaHS group, and IS groups with and without NaHS treatment. Serum was obtained to determine corticosterone (CORT), total antioxidant capacity (TAC), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) levels. Lung H2S, nitric oxide (NO), inducible nitric oxide synthase (iNOS), and malondialdehyde (MDA) levels were measured. Lung expressions of H2S synthesizing enzymes and Western blot analysis of nuclear factor erythroid 2-related factor 2 (Nrf2) and hypoxia-inducible factor 1 alpha (HIF 1α) were estimated. Histopathological changes and immunohistochemical assessment of nuclear factor kappa B (NF-κB) and caspase-3 were also done. Pretreatment with NaHS led to marked histological protection from lung damage seen in IS rats. Furthermore, pretreatment with NaHS before IS protected lung H2S levels and expressions of H2S-synthesizing enzymes. Similarly, the levels of CORT, TNF-α, IL-10, MDA, TAC, NO, iNOS, HIF-1 α, and nuclear Nrf2 and expressions of NF-kB and caspase 3 were all maintained at near control levels in contrast to that in the IS rats. In conclusion, NaHS is protective against stress-induced lung injury due to its antioxidant, anti-inflammatory, anti-fibrotic, and antiapoptotic effects. Thus, NaHS can be used to minimize stress complications on lung.

The effect of hydrogen sulfide on ischemi̇a /reperfusion injury in an experimental testicular torsion model

Testicular torsion is an acute pediatric surgical emergency requiring rapid diagnosis to prevent the permanent ischaemic damage of the testicles. Hydrogen Sulfide (H2S) have shown to cure tissue damage and has a role in the prevention of I/R damage. We aimed to evaluate the effect of H2S in testicular torsion. Eighteen male, Wistar albino rats were divided into 3 groups. The sham group which is applied surgical stress. The ischemia/reperfusion group (I/R) which detorsion performed 1h later than testicular torsion application. I/R+NaHS treatment group, NaHS solution was injected intraperitoneally for 1 week. On the 7th day of the detorsion all left testes were fixed in Bouin solution and sent to Pathology Department for histopathological examination. All right testes were washed with normal saline, dried in a sterile way and stored in - 80°C deepfreeze up to the date of biochemical processes. Testicular tissues were obtained for the detection of myeloperoxidase (MPO), malondialdehyde (MDA), AOPP (advanced oxidation protein product) for oxidant markers and ferric reducing antioxidant power (FRAP) levels, superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) activities for antioxidant markers and histopathological exploration. The effects of NaHs administration on oxidation were evaluated by determination of testicular MPO, MDA and AOPP levels. Increased testicular MPO (58.6%) activity was observed in the I/R group compared to the sham group. Following NaHS treatment, MPO (26.7%) activity was significantly decreased in rats exposed to I/R injury (Figure1). MDA levels did not alter. Increases in AOPP (20.9%) levels were observed in the I/R group. NaHS treatment resulted in significant decreases in AOPP (25.1%) levels in testes tissues of rats exposed to I/R injury. The effects of NaHS treatment on antioxidant system FRAP, SOD, GSH and GSH-Px activities were evaluated. GSH levels were significantly increased in the IR+NaHS group compared to the I/R group. In histopathological examination degeneration of seminiferous tubules and spermatogenic cells were observed in the I/R group. After NaHS treatment, normal spermatogenic activity with many spermatozoa in the lumen of most seminiferous tubules were observed in the I/R injured rats. According to Johnsen's scoring (JS), the I/R group was significantly decreased compared to the sham group. JS values for the I/R+NaHS group were significantly increased compared to the I/R group. Our study supports that ischemia/reperfusion injury plays an important role in the testicular torsion injury, and it is a pioneer study showing that H₂S may have a potential for therapeutic effect. The limitation of this work is this is an experimental study with limited number of animals. According to the results of our study, hydrogen sulfide treatment has beneficial effects on biochemical and histopathological results of testicular injury in testic torsion.

Hydrogen sulfide, a gaseous signaling molecule, elongates primary cilia on kidney tubular epithelial cells by activating extracellular signal-regulated kinase.

Primary cilia on kidney tubular cells play crucial roles in maintaining structure and physiological function. Emerging evidence indicates that the absence of primary cilia, and their length, are associated with kidney diseases. The length of primary cilia in kidney tubular epithelial cells depends, at least in part, on oxidative stress and extracellular signal-regulated kinase 1/2 (ERK) activation. Hydrogen sulfide (H2S) is involved in antioxidant systems and the ERK signaling pathway. Therefore, in this study, we investigated the role of H2S in primary cilia elongation and the downstream pathway. In cultured Madin-Darby Canine Kidney cells, the length of primary cilia gradually increased up to 4 days after the cells were grown to confluent monolayers. In addition, the expression of H2S-producing enzyme increased concomitantly with primary cilia length. Treatment with NaHS, an exogenous H2S donor, accelerated the elongation of primary cilia whereas DL-propargylglycine (a cystathionine γ-lyase inhibitor) and hydroxylamine (a cystathionine-β-synthase inhibitor) delayed their elongation. NaHS treatment increased ERK activation and Sec10 and Arl13b protein expression, both of which are involved in cilia formation and elongation. Treatment with U0126, an ERK inhibitor, delayed elongation of primary cilia and blocked the effect of NaHS-mediated primary cilia elongation and Sec10 and Arl13b upregulation. Finally, we also found that H2S accelerated primary cilia elongation after ischemic kidney injury. These results indicate that H2S lengthens primary cilia through ERK activation and a consequent increase in Sec10 and Arl13b expression, suggesting that H2S and its downstream targets could be novel molecular targets for regulating primary cilia.

Hydrogen Sulphide Treatment Prevents Renal Ischemia-Reperfusion Injury by Inhibiting the Expression of ICAM-1 and NF-kB Concentration in Normotensive and Hypertensive Rats.

Our main objective was to investigate the effect of chronic administration of hydrogen sulphide donor (sodium hydrosulphide) on the expression of intercellular adhesion molecule-1 (ICAM-1) and concentration of nuclear factor-kappa B (NF-kB) in a renal ischemia-reperfusion injury (IRI) model of WKY and L-nitro-arginine-methyl-ester (L-NAME)-induced hypertensive rats. Sodium hydrosulphide (NaHS) was administered intraperitoneally (i.p.) for 35 days while cystathionine gamma lyase (CSE) inhibitor dL-propargylglycine (PAG) was administered at a single dose of 50 mg/kg. Animals were anesthetised using sodium pentobarbitone (60 mg/kg) and then prepared to induce renal ischemia by clamping the left renal artery for 30 min followed by 3 h of reperfusion. Pre-treatment with NaHS improved the renal functional parameters in both WKY and L-NAME-induced hypertensive rats along with reduction of blood pressure in hypertensive groups. Oxidative stress markers like malondialdehyde (MDA), total superoxide dismutase (T-SOD) and glutathione (GSH) were also improved by NaHS treatment following renal IRI. Levels of ICAM-1 and NF-kB concentration were reduced by chronic treatment with NaHS and increased by PAG administration after renal IRI in plasma and kidney. Treatment with NaHS improved tubular morphology and glomerulus hypertrophy. Pre-treatment with NaHS reduced the degree of renal IRI by potentiating its antioxidant and anti-inflammatory mechanism, as evidenced by decreased NF-kB concentration and downregulation of ICAM-1 expression.

Hydrogen sulfide prevents arterial medial calcification in rats with diabetic nephropathy

BackgroundArterial medial calcification (AMC) is associated with a high incidence of cardiovascular risk in patients with type 2 diabetes and chronic kidney disease. Here, we tested whether hydrogen sulfide (H2S) can prevent AMC in rats with diabetic nephropathy (DN).MethodsDN was induced by a single injection of streptozotocin and high-fat diet (45% kcal as fat) containing 0.75% adenine in Sprague–Dawley rats for 8 weeks.ResultsRats with DN displayed obvious calcification in aorta, and this was significantly alleviated by Sodium Hydrosulfide (NaHS, a H2S donor, 50 μmol/kg/day for 8 weeks) treatment through decreasing calcium and phosphorus content, ALP activity and calcium deposition in aorta. Interestingly, the main endogenous H2S generating enzyme activity and protein expression of cystathionine-γ-lyase (CSE) were largely reduced in the arterial wall of DN rats. Exogenous NaHS treatment restored CSE activity and its expression, inhibited aortic osteogenic transformation by upregulating phenotypic markers of smooth muscle cells SMα-actin and SM22α, and downregulating core binding factor α-1 (Cbfα-1, a key factor for bone formation), protein expressions in rats with DN when compared to the control group. NaHS administration also significantly reduced Stat3 activation, cathepsin S (CAS) activity and TGF-β1 protein level, and improved aortic elastin expression.ConclusionsH2S may have a clinical significance for treating AMC in people with DN by reducing Stat3 activation, CAS activity, TGF-β1 level and increasing local elastin level.

Sodium Hydrosulfide Induces Resistance Against Penicillium expansum in Apples by Regulating Hydrogen Peroxide and Nitric Oxide Activation of Phenylpropanoid Metabolism.

As a multifunctional signaling molecule, hydrogen sulfide (H2S) has been reported to induce plant responses to a variety of abiotic stresses. However, there are no reports on H2S treatment inducing resistance in apples against Penicillium expansum, a biotic factor, and its possible mechanism of action. In this study, fumigating apples with 5 mM sodium hydrosulfide (NaHS), the exogenous donor of H2S, for 12 h reduced the diameter of lesions in fruit colonized by P. expansum. NaHS treatment markedly promoted the synthesis of endogenous H2S, hydrogen peroxide (H2O2), and nitrogen oxide (NO). In vivo NaHS treatment enhanced the activities of phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, p-coumarate:coenzyme A ligase isoenzymes, caffeoyl-CoA-O-methyltransferase, caffeic acid-O-methyltransferase, ferulic acid-5-hydroxylase, cinnamyl-CoA reductase, and cinnamyl-alcohol dehydrogenase. The treatment also facilitated the production of specific phenolic acids, such as cinnamic acid, p-coumaric acid, caffeic acid, ferulic acid, and sinapic acid; total phenolic compounds; p-coumaryl alcohol; coniferyl alcohol; sinapyl alcohol; and lignin. NaHS treatment induced resistance against P. expansum in apples through H2O2- and NO-mediated activation of phenylpropanoid metabolism.

Exercise training ameliorates early diabetic kidney injury by regulating the H2 S/SIRT1/p53 pathway.

Exercise training exerts protective effects against diabetic nephropathy. This study aimed to investigate whether exercise training could attenuate diabetic renal injury via regulating endogenous hydrogen sulfide (H2 S) production. First, C57BL/6 mice were allocated into the control, diabetes, exercise, and diabetes+exercise groups. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ). Treadmill exercise continued for four weeks. Second, mice was allocated into the control, diabetes, H2 S and diabetes+H2 S groups. H2 S donor sodium hydrosulfide (NaHS) was intraperitoneally injected once daily for four weeks. STZ-induced diabetic mice exhibited glomerular hypertrophy, tissue fibrosis and increased urine albumin levels, urine protein- and albumin-to-creatinine ratios, which were relieved by exercise training. Diabetic renal injury was associated with apoptotic cell death, as evidenced by the enhanced caspase-3 activity, the increased TdT-mediated dUTP nick-end labeling -positive cells and the reduced expression of anti-apoptotic proteins, all of which were attenuated by exercise training. Exercise training enhanced renal sirtuin 1 (SIRT1) expression in diabetic mice, accompanied by an inhibition of the p53-#ediated pro-apoptotic pathway. Furthermore, exercise training restored the STZ-mediated downregulation of cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) and the reduced renal H2 S production. NaHS treatment restored SIRT1 expression, inhibited the p53-mediated pro-apoptotic pathway and attenuated diabetes-associated apoptosis and renal injury. In high glucose-treated MPC5 podocytes, NaHS treatment inhibited the p53-mediated pro-apoptotic pathway and podocyte apoptosis in a SIRT1-dependent manner. Collectively, exercise training upregulated CBS/CSE expression and enhanced the endogenous H2 S production in renal tissues, thereby contributing to the modulation of the SIRT1/p53 apoptosis pathway and improvement of diabetic nephropathy.

Molybdenum and hydrogen sulfide synergistically mitigate arsenic toxicity by modulating defense system, nitrogen and cysteine assimilation in faba bean (Vicia faba L.) seedlings

Hydrogen sulfide (H2S) has emerged as a potential gasotransmitter in plants with a beneficial role in stress amelioration. Despite the various known functions of H2S in plants, not much information is available to explain the associative role of molybdenum (Mo) and hydrogen sulfide (H2S) signaling in plants under arsenic toxicity. In view to address such lacunae in our understanding of the integrative roles of these biomolecules, the present work attempts to decipher the roles of Mo and H2S in mitigation of arsenate (AsV) toxicity in faba bean (Vicia faba L.) seedlings. AsV-stressed seedlings supplemented with exogenous Mo and/or NaHS treatments (H2S donor) showed resilience to AsV toxicity manifested by reduction of apoptosis, reactive oxygen species (ROS) content, down-regulation of NADPH oxidase and GOase activity followed by upregulation of antioxidative enzymes in leaves. Fluorescent localization of ROS in roots reveals changes in its intensity and spatial distribution in response to MO and NaHS supplementation during AsV stress. Under AsV toxicity conditions, seedlings subjected to Mo + NaHS showed an increased rate of nitrogen metabolism evident by elevation in nitrate reductase, nitrite reductase and glutamine synthetase activity. Furthermore, the application of Mo and NaHS in combination positively upregulates cysteine and hydrogen sulfide biosynthesis in the absence and presence of AsV stress. Mo plus NaHS-supplemented seedlings exposed to AsV toxicity showed a substantial reduction in oxidative stress manifested by reduced ELKG, lowered MDA content and higher accumulation of proline in leaves. Taken together, the present findings provide substantial evidence on the synergetic role of Mo and H2S in mitigating AsV stress in faba bean seedlings. Thus, the application of Mo and NaHS reveals their agronomic importance to encounter heavy metal stress for management of various food crops.

CSE-Derived H2S Inhibits Reactive Astrocytes Proliferation and Promotes Neural Functional Recovery after Cerebral Ischemia/Reperfusion Injury in Mice Via Inhibition of RhoA/ROCK2 Pathway.

The effect of cystathionine-γ-lyase (CSE)-derived hydrogen sulfide (H2S) on the reactive proliferation of astrocytes and neural functional recovery over 30 d after acute cerebral ischemia and reperfusion (I/R) was determined by applying wild-type (WT) and CSE knockout (KO) mice. The changes of glial fibrillary acidic protein (GFAP) expression in hippocampal tissues was tested. Besides, we assessed the changes of mice spatial learning memory ability, neuronal damage, RhoA, Rho kinase 2 (ROCK2), and myelin basic protein (MBP) expressions in hippocampal tissues. The results revealed that cerebral I/R resulted in obvious increase of GFAP expression in hippocampal tissues. Besides, we found the neuronal damage, learning, and memory deficits of mice induced by cerebral I/R as well as revealed the upregulation of RhoA and ROCK2 expressions and reduced MBP expression in hipppcampal tissues of mice following cerebral I/R. Not surprisingly, the GFAP expression and cerebral injury as well as the upregulation of the RhoA/ROCK2 pathway were more remarkable in CSE KO mice, compared with those in WT mice over 30 d following acute cerebral I/R, which could be blocked by NaHS treatment, a donor of exogenous H2S. In addition, the ROCK inhibitor Fasudil also inhibited the reactive proliferation of astrocytes and ameliorated the recovery of neuronal function over 30 d after cerebral I/R. For the purpose of further confirmation of the role of H2S on the astrocytes proliferation following cerebral I/R, the immunofluorescence double staining: bromodeoxyuridine (BrdU) and GFAP was evaluated. There was a marked upregulation of BrdU-labeled cells coexpressed with GFAP in hippocampal tissues at 30 d after acute cerebral I/R; however, the increment of astrocytes proliferation could be ameliorated by both NaHS and Fasudil. These findings indicated that CSE-derived H2S could inhibit the reactive proliferation of astrocytes and promote the recovery of mice neural functional deficits induced by a cerebral I/R injury via inhibition of the RhoA/ROCK2 signal pathway.

H2S improves growth of tomato seedlings involving the MAPK signaling

As a signal molecule, hydrogen sulfide (H2S) plays an important role in plant growth. In this study, the effect of different concentration of H2S donor, sodium hydrosulfide (NaHS), on growth of tomato seedlings was tested in hydroponic culture. The effects of 50 μM NaHS on tomato seedlings were not significant, compared with the control. After 100 μM NaHS treatment, plant height, shoot fresh weight and dry weight increased significantly. However, 300, 500 and 1000 μM NaHS had inhibitory effect on tomato seedlings, especially 1000 μM NaHS. 100 μM NaHS could increase chlorophyll a, chlorophyll b, and total chlorophyll content, stomatal conductance, net photosynthetic rate, transpiration rate and intercellular CO2 concentration. The activities of nitrate reductase, glutamate synthase and glutamine synthase, and L-cysteine demethylase (LCD), and nitrate content, were increased significantly after 100 μM NaHS treatment. The microarray results showed that 9122 genes changed after treated with 100 μM NaHS for 3 h, and 430 genes changed more than 2 times. These genes were grouped into metabolism, signal transduction, stress resistance, transcription factors, transport, protein synthesis and degradation, hormone response, and cell wall related genes in tomato seedlings. RT-PCR analysis showed that the expression of some MAPK family genes changed significantly when exogenous NaHS was added, especially SlMAPK3 and SlMAPK13. The MAPK inhibitor PD98059 reduce the LCD activity and H2S content in tomato seedlings, which indicated that MAPK signaling pathway was involved in the process of H2S promoting tomato seedling growth.

Exogenous H2S Protects Colon Cells in Ulcerative Colitis by Inhibiting NLRP3 and Activating Autophagy.

Hydrogen sulfide (H2S) has been reported to participate in intestinal mucosal defense and repair. However, the precise regulatory mechanisms of H2S in ulcerative colitis (UC) remain unclear. We explored the effects of sodium hydrosulfide (NaHS), a donor of H2S, in dextran sulfate sodium (DSS)-induced colitis in rats. The pathologic features were determined by analyzing the hematoxylin and eosin-stained samples. Interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), and myeloperoxidase (MPO) levels were determined using ELISA. The presence of cystathionine-γ-lyase (CSE) and light chain 3B (LC3B) were determined using immunohistochemical and immunofluorescence (IF) approaches, respectively. Next, we investigated the effects of NaHS in lipopolysaccharide (LPS)-stimulated human colonic smooth muscle cells (H2940). The level of reactive oxygen species (ROS) was determined using IF. NOD-like receptor 3 (NLRP3) and CSE were detected using western blot and quantitative real-time polymerase chain reaction. Autophagy was determined using western blot, IF, and electron microscopy. NaHS treatment considerably diminished colitis-induced histological injury and proinflammatory cytokine expressions. MPO, CSE, and H2S were downregulated, whereas LC3B was upregulated after NaHS administration in colitic rats. NaHS remarkably attenuated the levels of ROS, CSE, and NLRP3 in LPS-stimulated cells and enhanced autophagy, as was revealed by increased LC3-II-to-LC3-I ratio, elevated LC3, and decreased p62. Importantly, NaHS promoted autophagosome formation in LPS-treated cells. Exogenous H2S ameliorates intestinal injury by downregulating inflammation and activation of autophagy, suggesting the potential of NaHS as a therapeutic agent for UC.

Stomatal regulation: Role of H2S-induced persulfidation in ABA signaling

Stomatal regulation: Role of H2S-induced persulfidation in ABA signaling

Pharmacological Evaluation of Beta‐Hydroxifosphocarnitine in No‐Alcoholic Steatohepatitis Induced in Rats

Aim Evaluate the pharmacological effect of Β-Hydroxyphosphocarnitine (β-HFC) on non-alcoholic steatohepatitis induced in rats. Introduction Non-alcoholic steatohepatitis (NASH) is a disease with high prevalence, usually accompanied on metabolic syndrome. Currently the treatment to NASH is pioglitazone. However, it is reported that pioglitazone causes several side effects reason to stop treatment. L-carnitine has pharmacological effect in the metabolic syndrome, but it is poorly absorbed. The β-HFC is an analogue of L-carnitine that has shown effect on metabolic syndrome and maintaining its pharmacological properties. Method The NASH model was developed in male Wistar rats by ingesting carbonated drink as a source of fructose, coconut oil and carbon tetrachloride. 3 groups were formed with NASH (n = 8); NASH, NASH -β-HFC and NASH-pioglitazone. Another 3 groups of healthy rats (n = 8); control, control-β-HFC and control-pioglitazone. The effect of β-HFC in NASH rats was evaluated by histological sections of the liver with Masson's hematoxylin eosin and trichromic staining. Quantification of liver enzymes, triglycerides, cholesterol and glucose in serum by spectrophotometric kit. Insulin levels were determined by Elisa. Results We previously reported that β-HFC reduced serum triglyceride levels in NASH rats. The levels of glucose, insulin and nitric oxide (NO) were quantified and are the ones that we will report on this occasion. Insulin levels were low in the NASH rats even with the β-HFC treatment compared to the control rats, (25.9 ± 6, 30.0 ± 10 and 121.1 ± 69.5 ng / ml respectively). The pancreas can be damaged by excess fatty acids (FA), altering insulin production. No significant difference was observed in glucose levels between the control and NASH groups (123.2 ± 50.6 and 134.3 ± 19.1 mg/dL respectively). High glucose levels would be expected in NASH rats, however, since insulin resistance is not observed, it could justify that glucose is not elevated. NO is a source of production of reactive species. The rats with NASH showed higher levels of NO than those of the control group (0.054, 0.01 µM / µl respectively). β-HFC was shown to reduce NO levels in rats with NASH (0.03µM / µl). β-HFC enhances the fatty acid oxidation process which can result in decreased production of reactive species. Conclusion: β-HFC enhances key alterations for NASH progression such as DNL and NO production. Conclusion: β-HFC promises to be effective for the treatment of NAHS by improving key alterations for the progression of NASH such as DNL and NO production.

Hydrogen Sulfide Subchronic Treatment Improves Hypertension Induced by Traumatic Brain Injury in Rats through Vasopressor Sympathetic Outflow Inhibition.

Traumatic brain injury (TBI) represents a critical public health problem around the world. To date, there are no accurate therapeutic approaches for the management of cardiovascular impairments induce by TBI. In this regard, hydrogen sulfide (H2S), a novel gasotransmitter, has been proposed as a neuro- and cardioprotective molecule. This study was designed to determine the effect of subchronic management with sodium hydrosulfide (NaHS) on hemodynamic, vasopressor sympathetic outflow and sensorimotor alterations produced by TBI. Animals underwent a lateral fluid percussion injury, and changes in hemodynamic variables were measured by pletismographic methods. In addition, vasopressor sympathetic outflow was assessed by a pithed rat model. Last, sensorimotor impairments were evaluated by neuroscore test and beam-walking test. At seven, 14, 21, and 28 days after moderate-severe TBI, the animals showed: (1) a decrease on sensorimotor function in the neuroscore test and beam-walking test; (2) an increase in heart rate, systolic, diastolic, and mean blood pressure; (3) progressive sympathetic hyperactivity; and (4) a decrease in vasopressor responses induced by noradrenaline (α1/2-adrenoceptors agonist) and UK 14,304 (selective α2-adrenoceptor agonist). Interestingly, intraperitoneal daily injections of NaHS, an H2S donor (3.1 and 5.6 mg/kg), during seven days after TBI prevented the development of the impairments in hemodynamic variables, which were similar to those obtained in sham animals. Moreover, NaHS treatment prevented the sympathetic hyperactivity and decreased noradrenaline-induced vasopressor responses. No effects on sensorimotor dysfunction were observed, however. Taken together, our results suggest that H2S ameliorates the hemodynamic and sympathetic system impairments observed after TBI.

NaHS Alleviated Cell Apoptosis and Mitochondrial Dysfunction in Remote Lung Tissue after Renal Ischemia and Reperfusion via Nrf2 Activation-Mediated NLRP3 Pathway Inhibition.

Objective Acute kidney injury (AKI) is a common and severe complication in critically ill patients, often caused by renal ischemia-reperfusion (RIR). Previous studies have confirmed that lung injury, rather than renal injury, is one of the leading causes of AKI-induced death. The pathophysiological mechanisms of acute lung injury (ALI) resulting from AKI are very complex and remain unclear. In the present study, we aimed to explore the protective effects and potential mechanism of sodium hydrosulfide (NaHS) on lung injury in RIR mice. Methods The RIR model was established in wild-type and Nrf2−/− mice. Different groups of mice were treated with NaHS and MCC950. Lung tissues were harvested to detect lung injury, mitochondrial function, cell apoptosis, the NLRP3 inflammasome, and Nrf2 pathway-related molecules. Results RIR led to a deterioration in lung histology, the wet/dry weight ratio, PaO2/FiO2, and mitochondrial function, in addition to stimulating the activation of the NLRP3 and Nrf2 pathways. MCC950 alleviated mitochondrial dysfunction, lung apoptosis, and histology injury in the lungs after RIR. NaHS treatment markedly improved the lung histological scores, the wet/dry weight ratio, bronchoalveolar lavage fluid (BALF) cell counts, BALF neutrophil counts, BALF neutrophil elastase activity, BALF protein concentration, PaO2/FiO2, mitochondrial morphology, the red/green fluorescence intensity that indicates changes in mitochondrial membrane potential, respiratory control rate (RCR), ATP, reactive oxygen species (ROS) release, and cell apoptosis via Nrf2-mediated NLRP3 pathway inhibition. Conclusion NaHS protected against RIR-induced lung injury, mitochondrial dysfunction, and inflammation, which is associated with Nrf2 activation-mediated NLRP3 pathway inhibition.

Hydrogen sulfide regulates hippocampal neuron excitability via S-sulfhydration of Kv2.1

Hydrogen sulfide (H2S) is gaining interest as a mammalian signalling molecule with wide ranging effects. S-sulfhydration is one mechanism that is emerging as a key post translational modification through which H2S acts. Ion channels and neuronal receptors are key target proteins for S-sulfhydration and this can influence a range of neuronal functions. Voltage-gated K+ channels, including Kv2.1, are fundamental components of neuronal excitability. Here, we show that both recombinant and native rat Kv2.1 channels are inhibited by the H2S donors, NaHS and GYY4137. Biochemical investigations revealed that NaHS treatment leads to S-sulfhydration of the full length wild type Kv2.1 protein which was absent (as was functional regulation by H2S) in the C73A mutant form of the channel. Functional experiments utilising primary rat hippocampal neurons indicated that NaHS augments action potential firing and thereby increases neuronal excitability. These studies highlight an important role for H2S in shaping cellular excitability through S-sulfhydration of Kv2.1 at C73 within the central nervous system.

SODIUM HYDROGEN SULFIDE IMPROVES RENOVASCULAR HYPERTENSION-INDUCED MALADAPTIVE CARDIAC REMODELING AND DIASTOLIC DYSFUNCTION BY REDUCING OXIDATIVE STRESS AND MMP ACTIVITY

Objective: To evaluate whether treatment with sodium hydrogen sulfide (NaHS) improves cardiac function and remodeling of two kidney-one clip (2K-1C) rats through inhibition of matrix metalloproteinase (MMP) activity and oxidative stress. Design and method: 2K-1C hypertension was surgically induced in male Wistar rats (200 g; ethics commitee #118/2019) by an implantation of a silver clip (0.2 mm) in the left renal artery. Control rats (2K) was submitted to laparotomy. SBP and echocardiograph were registered. The NaHS (0.24 mg/Kg/day; i.p.) treatment or vehicle (0.6% DMSO in saline; i.p.) was initiated in the second week after the surgeries and lasted for 28 days. Hearts were collected and western blotting to MMP-2, Nox2 and Nox4 were performed in the left ventricles. MMPs localization and activity were also evaluated by in situ zymography. Reactive oxygen species (ROS) were investigated by dihydroethidium (10 μM). Data are presented as mean ± S.E.M. of different experimental n. Two-way ANOVA with Bonferroni correction was used (P < 0.05). Results: SBP values of 2K-1C (165.47 ± 3.98mmHg, n = 12) were increased compared to 2K rats (133.28 ± 2.48mmHg, n = 10; P < 0.0001) in the first week after the surgery. NaHS treatment decreased SBP of 2K-1C NaHS (155.60 ± 5.72mmHg, n = 5) compared to 2K-1C vehicle (202.13 ± 5.57mmHg, n = 7; P = 0.0003). Isovolumetric relaxation time analysis showed increases in values from 2K-1C rats (40.31 ± 2.97ms, n = 7) and NaHS treatment (28.04 ± 1.05ms, n = 5) normalized this parameter (P = 0.0018). 2K-1C rats (2.30 ± 0.11 mm, n = 7) presented increased interventricular septum thickness and NaHS treatment (1.99 ± 0.0 mm, n = 5) reduced this parameter (P = 0.028). MMP-2, Nox2 or Nox4 expression was not different in all groups. The gelatinolytic activity of MMPs was higher in 2K-1C vehicle rats (18,029.31 ± 1,225.42U, n = 3) and NaHS (12,092.70 ± 868.22U, n = 3) reduced it (P = 0.021). In 2K-1C rats (33,868.95 ± 2,103.74U, n = 3), ROS were higher compared to control and NaHS treatment (22,019.50 ± 914.08 U, n = 3) decreased this value (P = 0.0021). Conclusions: Our results showed beneficial effects of NaHS treatment in 2K-1C rats, thus leading to SBP decrease, cardiac function improvement and reduction in both MMPs activity and oxidative stress.