Gasotransmitters: Potential Therapeutic Molecules of Fibrotic Diseases.

Fibrosis is defined as the pathological progress of excessive extracellular matrix (ECM), such as collagen, fibronectin, and elastin deposition, as the regenerative capacity of cells cannot satisfy the dynamic repair of chronic damage. The well-known features of tissue fibrosis are characterized as the presence of excessive activated and proliferated fibroblasts and the differentiation of fibroblasts into myofibroblasts, and epithelial cells undergo the epithelial-mesenchymal transition (EMT) to expand the number of fibroblasts and myofibroblasts thereby driving fibrogenesis. In terms of mechanism, during the process of fibrosis, the activations of the TGF-ß signaling pathway, oxidative stress, cellular senescence, and inflammatory response play crucial roles in the activation and proliferation of fibroblasts to generate ECM. The deaths due to severe fibrosis account for almost half of the total deaths from various diseases, and few treatment strategies are available for the prevention of fibrosis as yet. Recently, numerous studies demonstrated that three well-defined bioactive gasotransmitters, including nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S), generally exhibited anti-inflammatory, antioxidative, antiapoptotic, and antiproliferative properties. Besides these effects, a number of studies have reported that low-dose exogenous and endogenous gasotransmitters can delay and interfere with the occurrence and development of fibrotic diseases, including myocardial fibrosis, idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis, diabetic diaphragm fibrosis, and peritoneal fibrosis. Furthermore, in animal and clinical experiments, the inhalation of low-dose exogenous gas and intraperitoneal injection of gaseous donors, such as SNAP, CINOD, CORM, SAC, and NaHS, showed a significant therapeutic effect on the inhibition of fibrosis through modulating the TGF-ß signaling pathway, attenuating oxidative stress and inflammatory response, and delaying the cellular senescence, while promoting the process of autophagy. In this review, we first demonstrate and summarize the therapeutic effects of gasotransmitters on diverse fibrotic diseases and highlight their molecular mechanisms in the process and development of fibrosis.

Recent Development of Hydrogen Sulfide Therapeutics in the Treatment of Cardiovascular Diseases.

Hydrogen sulfide (H2S), as one of the endogenous gasotransmitters, has shown great potential in treating cardiovascular diseases (CVDs). H2S plays a protective role in CVDs by removing reactive oxygen species (ROS), promoting vasodilation, inhibiting myocardial hypertrophy, preventing thrombosis, and protecting mitochondria. However, there still exist some problems for H2S as drugs such as challenging delivery, uncontrollable release rate, and other drug developability issues. Addressing these problems, the prodrug strategy shows great potential. Therefore, a key issue on the H2S-based therapeutics is developing appropriate H2S prodrugs. In this review, we mainly discussed the mechanism of H2S against CVDs and reviewed the cardiovascular effects of current H2S prodrugs.

FRET Modulated Signaling: A Versatile Strategy to Construct Photoelectrochemical Microsensors for In Vivo Analysis.

Microelectrode-based electrochemical (EC) and photoelectrochemical (PEC) sensors are promising candidates for in vivo analysis of biologically important chemicals. However, limited selectivity in complicated biological systems and poor adaptability to electrochemically non-active species restrained their applications. Herein, we propose the concept of modulating the PEC output by a fluorescence resonance energy transfer (FRET) process. The emission of energy donor was dependent on the concentration of target SO2 , which in turn served as the modulator of the photocurrent signal of the photoactive material. The employment of optical modulation circumvented the problem of selectivity, and the as-fabricated PEC microelectrode showed good stability and reproducibility in vivo. It can monitor fluctuations of SO2 levels in brains of rat models of cerebral ischemia-reperfusion and febrile seizure. More significantly, such a FRET modulated signaling strategy can be extended to diverse analytes.

Porous materials as carriers of gasotransmitters towards gas biology and therapeutic applications.

The discovery of NO, CO, and H2S as gasotransmitters and their beneficial role in multiple physiological functions opened an era of research devoted to exogenously delivering them as therapeutic agents. However, the gaseous nature of these molecules demands new forms of administration that enable one to control the location, dosage and timing of their delivery. Porous materials are among the most suitable scaffolds to store, deliver and release gasotransmitters due to their high surface area, tunable composition and reactivity. This review highlights the strategies employed to load and release gasotransmitters from different kinds of porous materials, including zeolites, mesoporous silica, metal-organic frameworks and protein assemblies.

The Roles of NO and H2S in Sperm Biology: Recent Advances and New Perspectives.

After being historically considered as noxious agents, nitric oxide (NO) and hydrogen sulfide (H2S) are now listed as gasotransmitters, gaseous molecules that play a key role in a variety of cellular functions. Both NO and H2S are endogenously produced, enzymatically or non-enzymatically, and interact with each other in a range of cells and tissues. In spite of the great advances achieved in recent decades in other biological systems, knowledge about H2S function and interactions with NO in sperm biology is in its infancy. Here, we aim to provide an update on the importance of these molecules in the physiology of the male gamete. Special emphasis is given to the most recent advances in the metabolism, mechanisms of action, and effects (both physiological and pathophysiological) of these gasotransmitters. This manuscript also illustrates the physiological implications of NO and H2S observed in other cell types, which might be important for sperm function. The relevance of these gasotransmitters to several signaling pathways within sperm cells highlights their potential use for the improvement and successful application of assisted reproductive technologies.

Sulfane sulfur - new findings on an old topic.

Sulfane sulfur is a divalent sulfur atom bonded to another sulfur which is very reactive and labile. Compounds containing this reactive sulfur include persulfides, polysulfides, thiosulfate, thiosulfinates, polythionates, and elemental sulfur. Sulfane sulfur appears in a number of biologically important compounds, including thiocysteine, thiocystine and thiotaurine, products of the cysteine metabolism, as well as glutathione persulfide. Sulfane sulfur compounds can modify cysteine residues in proteins via an S-sulfhydration reaction to produce protein persulfides. It has been also postulated that cysteine persulfides can be incorporated into proteins during translation. Recently, the sulfane sulfur compounds, especially the persulfides and polysulfides, have attracted increasing interest due to their regulatory and antioxidant properties. Compounds containing sulfane sulfur are also regarded as a form of H2S storage, which can easily release this gasotransmitter in response to biological signals. Both reactive sulfur species (H2S and sulfane sulfur) always coexist in biological systems. This review is focused on new findings in the field of sulfane sulfur's biological role, and disruption of its level in some patho/physiological conditions. A few sulfane sulfur donors with potential applications are presented. In recent years, in parallel to increasing interest in biological importance of sulfane sulfur, new analytical methods have been developed for sensitive and reliable determination of its level in the cells and tissues.

Gasotransmitter Regulation of Phosphatase Activity in Live Cells Studied by Three-Channel Imaging Correlation.

Enzyme activity in live cells is dynamically regulated by small-molecule transmitters for maintaining normal physiological functions. A few probes have been devised to measure intracellular enzyme activities by fluorescent imaging, but the study of the regulation of enzyme activity via gasotransmitters in situ remains a long-standing challenge. Herein, we report a three-channel imaging correlation by a single dual-reactive fluorescent probe to measure the dependence of phosphatase activity on the H2 S level in cells. The two sites of the probe reactive to H2 S and phosphatase individually produce blue and green fluorescent responses, respectively, and resonance energy transfer can be triggered by their coexistence. Fluorescent analysis based on the three-channel imaging correlation shows that cells have an ideal level of H2 S to promote phosphatase activity up to its maximum. Significantly, a slight deviation from this H2 S level leads to a sharp decrease of phosphatase activity. The discovery further strengthens our understanding of the importance of H2 S in cellular signaling and in various human diseases.

Colorimetric sensor for the detection of H2S and its application in molecular half-subtractor.

Azine based new colorimetric sensor 1 for the detection of gasotransmitter H2S has been reported. Sensor 1 used to detect the H2S with a remarkable red shift of 105 nm in the absorption spectra with a colour change from light yellow to brown red. Importantly, rare example of azine derivative has been used as a colorimetric probe for H2S detection using deprotonation mechanism. H2S induced deprotonation of one of the -OH proton followed by a change in resonance of 1 is responsible for the ratiometric spectral and colour change. The detection response was quick and the LOD calculated as 18.2 µM. Sensor 1 was also explored in the detection of H2S in biological fluids such as human serum and mouse serum. Moreover, for the first time, we have shown the applicability of H2S for the construction of half subtractor molecular logic gate.

Hydrogen sulfide induced supramolecular self-assembly in living cells.

Here we developed a critical gasotransmitter (H2S) mediated reduction to induce supramolecular self-assembly in multiple living cell lines. Specifically, the reduction converted an azido group to an amine which allowed the formation of intramolecular hydrogen bonds. The hydrogen bonds planarized the assembling molecules and promoted the supramolecular self-assembly.

H2S Delivery from Aromatic Peptide Amphiphile Hydrogels.

Hydrogels are materials composed mostly of water that have found use in a wide variety of applications, including tissue engineering and regenerative medicine. Aromatic peptide amphiphiles can be designed to self-assemble in aqueous solution into one-dimensional aggregates that entangle to form hydrogels with very high water content (>99 wt. %). Here, we describe the synthesis of an aromatic peptide amphiphile designed to release hydrogen sulfide (H2S), a vital biological signaling gas with significant therapeutic potential. Peptide synthesis, purification, aliquotting, and procedures for measuring H2S release are detailed.

A review of hydrogen sulfide (H2S) donors: Chemistry and potential therapeutic applications.

Hydrogen sulfide (H2S) is a ubiquitous small gaseous signaling molecule, playing an important role in many physiological processes and joining nitric oxide and carbon monoxide in the group of signaling agents termed gasotransmitters. Endogenous concentrations of H2S are generally low, making it difficult to discern precise biological functions. As such, probing the physiological roles of H2S is aided by exogenous delivery of the gas in cell and animal studies. This need for an exogenous source of H2S provides a unique challenge for chemists to develop chemical tools that facilitate the study of H2S under biological conditions. Compounds that degrade in response to a specific trigger to release H2S, termed H2S donors, include a wide variety of functional groups and delivery systems, some of which mimic the tightly controlled endogenous production in response to specific, biologically relevant conditions. This review examines a variety of H2S donor systems classified by their H2S-releasing trigger as well as their H2S release profiles, byproducts, and potential therapeutic applications.

A two-photon excitable and ratiometric fluorogenic nitric oxide photoreleaser and its biological applications.

We report for the first time the development of a two-photon excitable NO photoreleaser, CNNO, for ratiometric imaging and tracking of NO release in live cells. CNNO exhibits the merits of spatiotemporal control in both the site-specific NO release in the selected cell culture region and the controllable vasodilation of mouse aorta ex vivo.

Iminothioethers as Hydrogen Sulfide Donors: From the Gasotransmitter Release to the Vascular Effects.

The gasotransmitter hydrogen sulfide (H2S) is an important tuner of the cardiovascular homeostasis, and its deficiency is etiologically associated with a number of cardiovascular diseases. Therefore, the research of original moieties able to release H2S represents a timely issue for drug discovery. In this work, we developed a collection of iminothioethers (ITEs), exhibiting H2S-releasing properties and producing vasorelaxing effects on rat aortic rings. Derivatives 4 and 11, selected as representative of slow and fast rate H2S donors, respectively, produced a complete recovery of the basal coronary flow, reverting the AngII-induced effects in isolated rat hearts. In addition, studies on human aortic smooth muscle cells (HASMCs) demonstrated membrane hyperpolarizing effects, well related to the intracellular generation of H2S. Taken together, the results obtained support ITEs 4 and 11 as new pharmacological tools, as well as effective and innovative H2S donors for cardiovascular drug discovery.

Spectroscopic investigation of the reaction of metallo-protoporphyrins with hydrogen sulfide.

Hydrogen sulfide (H2S) is the most recently discovered gasotransmitter molecule joining nitric oxide and carbon monoxide. In addition to being biologically important gases, these gasotransmitters also provide distinct modes of reactivity with biomimetic metal complexes. The majority of previous investigations on the reactivity of H2S with bioinorganic models have focused on Fe-based porphyrin systems, whereas investigations with other metals remains underinvestigated. To address this gap, we report here an examination of the reactions of H2S, HS-, and S8 with MgII, CuII, CoII, ZnII, CrII, SnIV, and MnII/III protoporphyrins.

Click and Release: A Chemical Strategy toward Developing Gasotransmitter Prodrugs by Using an Intramolecular Diels-Alder Reaction.

Prodrug strategies have been proven to be a very effective way of addressing delivery problems. Much of the chemistry in prodrug development relies on the ability to mask an appropriate functional group, which can be removed under appropriate conditions. However, developing organic prodrugs of gasotransmitters represent unique challenges. This is especially true with carbon monoxide, which does not have an easy "handle" for bioreversible derivatization. By taking advantage of an intramolecular Diels-Alder reaction, we have developed a prodrug strategy for preparations of organic CO prodrugs that are stable during synthesis and storage, and yet readily release CO with tunable release rates under near physiological conditions. The effectiveness of the CO prodrug system in delivering a sufficient quantity of CO for possible therapeutic applications has been studied using a cell culture anti-inflammatory assay and a colitis animal model. These studies fully demonstrate the proof of concept, and lay a strong foundation for further medicinal chemistry work in developing organic CO prodrugs.

Signaling of hydrogen sulfide and polysulfides.

It has been almost two decades since the first demonstration of hydrogen sulfide (H2S) as a physiological mediator of cognitive function and vascular tone. H2S is physiologically important because it protects various organs from ischemia-reperfusion injury besides regulating inflammation, oxygen sensing, cell growth, and senescence. The production, metabolism, and regulation of H2S have been studied extensively. H2S modulates target proteins through sulfhydration (or sulfuration) or by the reduction of cysteine disulfide bonds. A large number of novel H2S-donating compounds are being developed owing to the therapeutic potential of H2S. Recently, polysulfides, rather than H2S, have been identified as molecules that sulfhydrate (or sulfurate) their target proteins.

H2S preconditioning of human adipose tissue-derived stem cells increases their efficacy in an in vitro model of cell therapy for simulated ischemia.

AIMS: A major limitation of cell-based therapies for ischemia-reperfusion injury is the excessive loss of administered cells. We investigated whether H2S can improve the survival and efficacy of therapeutic cells in an in vitro model of cell-based therapy for simulated ischemia. MAIN METHODS: H9c2 rat cardiomyoblasts were exposed to oxygen-glucose deprivation and NaHS (3-30 µM) pretreated human adipose tissue derived stem cells (hASCs) were added after reoxygenization. Viability of both cell lines was assessed with flow cytometry after 24h. The effects of H2S on antioxidant defense, proliferation, AKT and ERK1/2 phosphorylation and mitochondrial activity were analyzed in hASCs. Proliferation was evaluated using propargylglycine, an inhibitor of endogenous H2S synthesis. KEY FINDINGS: NaHS pretreatment decreased the ratio of necrotic therapeutic cells by 41.8% in case of 3 µM NaHS and by 34.3% with 30 µM NaHS. The ratio of necrotic postischemic cardiomyocytes decreased by 35%, but only with the use of 3 µM NaHS. Antioxidant defense mechanisms and ERK-phosphorylation were enhanced after 3 µM NaHS treatment while AKT-phosphorylation was suppressed. NaHS dose-dependently increased the proliferation of hASCs while pretreatment with propargylglycine decreased it. SIGNIFICANCE: NaHS pretreatment can increase the survival of therapeutically used human adipose tissue-derived stem cells via increased antioxidant defense and improves the postischemic cardiac derived cells' survival as well. Proliferation of human adipose tissue-derived stem cells is enhanced by H2S. The underlying mechanisms involve enhanced ERK-phosphorylation and decreased AKT-phosphorylation. Pretreatment with NaHS may represent a simple pharmacological step that may enhance the efficacy of cell-based therapies.

Hydrogen sulfide releasing capacity of natural isothiocyanates: is it a reliable explanation for the multiple biological effects of Brassicaceae?

Hydrogen sulfide is an endogenous pleiotropic gasotransmitter, which mediates important physiological effects in the human body. Accordingly, an impaired production of endogenous hydrogen sulfide contributes to the pathogenesis of important disorders. To date, exogenous compounds, acting as hydrogen sulfide-releasing agents, are viewed as promising pharmacotherapeutic agents. In a recent report, the hydrogen sulfide-releasing properties of some synthetic aryl isothiocyanate derivatives have been reported, indicating that the isothiocyanate function can be viewed as a suitable slow hydrogen sulfide-releasing moiety, endowed with the pharmacological potential typical of this gasotransmitter. Many isothiocyanate derivatives (deriving from a myrosinase-mediated transformation of glucosinolates) are well-known secondary metabolites of plants belonging to the family Brassicaceae, a large botanical family comprising many edible species. The phytotherapeutic and nutraceutic usefulness of Brassicaceae in the prevention of important human diseases, such as cancer, neurodegenerative processes and cardiovascular diseases has been widely discussed in the scientific literature. Although these effects have been largely attributed to isothiocyanates, the exact mechanism of action is still unknown. In this experimental work, we aimed to investigate the possible hydrogen sulfide-releasing capacity of some important natural isothiocyanates, studying it in vitro by amperometric detection. Some of the tested natural isothiocyanates exhibited significant hydrogen sulfide release, leading us to hypothesize that hydrogen sulfide may be, at least in part, a relevant player accounting for several biological effects of Brassicaceae.

A new fluorescent probe for gasotransmitter H2S: high sensitivity, excellent selectivity, and a significant fluorescence off-on response.

A fluorescent off-on probe for H2S was exploited by coupling the azide-based strategy with the excited-state intramolecular proton transfer (ESIPT) sensing mechanism, which exhibits a considerably high fluorescence enhancement (1150-fold), an extremely low detection limit (0.78 nM), and a relatively fast response time (3-10 min) as well as excellent selectivity.

Emerging role of gasotransmitters in renal transplantation.

Once patients with kidney disease progress to end-stage renal failure, transplantation is the preferred option of treatment resulting in improved quality of life and reduced mortality compared to dialysis. Although 1-year survival has improved considerably, graft and patient survival in the long term have not been concurrent, and therefore new tools to improve long-term graft and patient survival are warranted. Over the past decades, the gasotransmitters nitric oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H2S) have emerged as potent cytoprotective mediators in various diseases. All three gasotransmitters are endogenously produced messenger molecules that possess vasodilatory, anti-apoptotic, anti-inflammatory and anti-oxidant properties by influencing an array of intracellular signaling processes. Although many regulatory functions of gasotransmitters have overlapping actions, differences have also been reported. In addition, crosstalk between NO, CO and H2S results in synergistic regulatory effects. Endogenous and exogenous manipulation of gasotransmitter levels modulates several processes involved in renal transplantation. This review focuses on mechanisms of gas-mediated cytoprotection and complex interactions between gasotransmitters in renal transplantation.