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Cysathionine γ-lyase (CSE) is a core enzyme for the synthesis of endogenous H
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Objective To observe the levels of serum reactive oxygen species (ROS) and hydrogen sulfide(H2S) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD), and nicotinamide adenine dinucleotide phosphate?reduced (NADPH) oxidase 4 (NOX4) and cystathionine?γ?lyase (CSE) in lung tissue of patients with stable chronic obstructive pulmonary disease (COPD). Methods (1) A total of 60 patients with AECOPD admitted to the Department of Respiratory Medicine at Ningxia Hui People′s Hospital from November 2015 to December 2016 were recruited. According to the results of pulmonary function and echocardiography, the participants were divided into AECOPD?related pulmonary hypertension (PH) group(A) and AECOPD non?PH group (B).Other 30 healthy subjects were selected as the control group (C).Serum ROS and H2S of group A, B and C were detected by enzyme?linked immunosorbent assay (ELISA).(2)The lung tissues of patients undergoing lobectomy for lung cancer from November 2012 to April 2017 were collected, who were divided into COPD?related PH group (D), COPD non?PH group (E) and negative control (F). The expression of NOX4 and CSE protein in lung tissue was detected by immunohistochemistry and the thickness of pulmonary arteriole wall was measured. Results (1)The serum ROS level in group A was higher than group B and C which were (613.52±69.66)IU/ml,(565.76±71.33)IU/ml, (294.63±60.39)IU/ml, respectively with that in group B higher than that in group C (P<0.05). Serum H2S level in group A was lower than group B and C, with that in group B lower than group C [(18.59±5.50) nmol/ml, (20.49±4.97) nmol/ml, (38.03±4.43) nmol/ml, respectively P<0.05]. ROS level was positively correlated with pulmonary systolic pressure (PASP) (r=0.59, P<0.05), H2S level was negatively correlated with PASP(r=-0.62, P<0.05).(2)The lung tissue expression of NOX4 in group D was higher than group E and F (P<0.05), which were 0.08±0.01,0.06±0.01,0.03±0.01, respectively,while the level of NOX4 in group E was higher than group F (P<0.05). The expression of CSE between group D, E and F were all significantly different (P<0.05),which were 0.03±0.01, 0.07±0.02,0.12±0.02, respectively.(3)Smooth muscle thickness of pulmonary arterioles as a percentage of vascular diameter (WT%) between group D, E and F was all different(P<0.05), which were (40.58±6.63)%,(36.87±5.60)%,(31.27±6.24)%, respectively; so was smooth muscle area of pulmonary arterioles as a percentage of total vascular area(WA% ) with (32.33 ± 6.27)% , (30.20±5.28)%, (25.20±4.31)%, respectively (P<0.05). (4)The expression of NOX4 was positively correlated with WT% and WA% , r was 0.81 and 0.66, respectively (P<0.05). The expression of CSE was negatively correlated with WT% and WA%, r was-0.55 and-0.39 respectively (P<0.05). Conclusions NOX4/ROS and CSE/H2S signaling pathways may play an important role in the pathogenesis of COPD related PH.
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Objective To investigate the expression changes of the hydrogen sulfide synthases cystathionineγ-lyase (CSE), cystathionineβ-synthase (CBS), and 3-mercaptopyruvate sulfurtransferase (3-MST), after optic nerve crush (ONC) in rat the retina.Methods The rat model of ONC was established.Rats were divided into normal control, ONC, and sham control groups.Histopathologic changes in retina, the number of retinal ganglion cells (RGC) and retinal thickness of inner part (RTIP) were measured.The changes of CSE, CBS and 3-MST mRNA expression were detected with quantitative real-time PCR.Results The retinal histostructure was normal in normal controls and with minor changes in sham controls, respectively.Compared with sham group, significant retina damages were found in the ONC group:a time-dependent reduction of RGC number and RTIP.Expressions of CSE, CBS and 3-MST mRNA in rat retina were detected in normal control.Compared with normal controls, the expressions of CSE, CBS and 3-MST mRNA did not show any significant changes in the sham controls.Compared with sham controls, CBS mRNA expressions showed a time-dependent increase at 3 d, 7 d and 14 d after crush in the ONC group;CSE mRNA expressions increased to the peak at 3 d and then slightly reduced at 14 d after crush;3-MST mRNA expressions showed the trend of increase at 3 d and 7 d and then enhanced remarkably at 14 d after crush.Conclusion Hydrogen sulfide synthases CSE, CBS and3-MST expressions were up-regulated in rat retina following ONC, which may cause an increase in local endogenous hydrogen sulfide production in the retina and a compensatory protective effect.
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PURPOSE: Hydrogen sulfide (H2S) is an endogenous gaseous molecule with important physiological roles. It is synthesized from cysteine by cystathionine γ-lyase (CGL) and cystathionine β-synthase (CBS). The present study examined the benefits of exogenous H2S on renal ischemia reperfusion (IR) injury, as well as the effects of CGL or CBS inhibition. Furthermore, we elucidated the mechanism underlying the action of H2S in the kidneys. MATERIALS AND METHODS: Thirty male Sprague-Dawley rats were randomly assigned to five groups: a sham, renal IR control, sodium hydrosulfide (NaHS) treatment, H2S donor, and CGL or CBS inhibitor administration group. Levels of blood urea nitrogen (BUN), serum creatinine (Cr), renal tissue malondialdehyde (MDA), and superoxide dismutase (SOD) were estimated. Histological changes, apoptosis, and expression of mitogen-activated protein kinase (MAPK) family members (extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38) were also evaluated. RESULTS: NaHS attenuated serum BUN and Cr levels, as well as histological damage caused by renal IR injury. Administration of NaHS also reduced oxidative stress as evident from decreased MDA, preserved SOD, and reduced apoptotic cells. Additionally, NaHS prevented renal IR-induced MAPK phosphorylation. The CGL or CBS group showed increased MAPK family activity; however, there was no significant difference in the IR control group. CONCLUSION: Exogenous H2S can mitigate IR injury-led renal damage. The proposed beneficial effect of H2S is, in part, because of the anti-oxidative stress associated with modulation of the MAPK signaling pathways.
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Animals , Humans , Male , Rats , Apoptosis , Blood Urea Nitrogen , Creatinine , Cystathionine , Cysteine , Hydrogen Sulfide , Hydrogen , Ischemia , JNK Mitogen-Activated Protein Kinases , Kidney , Malondialdehyde , Oxidative Stress , Phosphorylation , Phosphotransferases , Protein Kinases , Rats, Sprague-Dawley , Reperfusion , Reperfusion Injury , Sodium , Superoxide Dismutase , Tissue DonorsABSTRACT
This study aimed to observe changes in the hydrogen sulfide (H2S) system in the blood and liver tissue of rats with hepatic cirrhosis at different stages by studying the effect of H2S on the course of hyperdynamic circulation in rats with hepatic cirrhosis.H2S concentration in the blood from the portal vein and inferior vena cava of hepatic cirrhosis rat model induced with carbon tetrachloride was detected on the 15th,30th,and 52nd day.The expression of cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) protein,and CBS and CSE mRNA in the liver was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR),respectively.The results indicated that H2S concentration in the blood from the portal vein and inferior vena cava of rats with hepatic cirrhosis was significantly lower than that in the control group.H2S was gradually decreased with the development of the disease and significantly lower in the blood from portal vein than in the blood of inferior vena cava at the mid-stage and the late stage groups.The expression levels of CBS and CSE protein,and CBS and CSE mRNA in the livers with hepatic cirrhosis at different stages were all higher than those in the control group,and the expression gradually increased with the development of the disease.The expression of CBS was lower than CSE in the same stages.The results indicated that the CSE mRNA was expressed predominantly in the cirrhosis groups as compared with CBS rnRNA.Among experimental rats,the H2S system has an important effect on the occurrence and development of hyperdynamic circulation in rats with hepatic cirrhosis.This finding adds to the literature by demonstrating that H2S protects vascular remodelling in the liver,and that CSE is indispensable in this process.
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Hydrogen sulphide is an endogenous inflammatory mediator produced by cystathionine-γ-lyase (CSE) in macrophages. To determine the role of H2S and macrophages in sepsis, we used small interference RNA (siRNA) to target the CSE gene and investigated its effect in a mouse model of sepsis. Cecal ligation puncture (CLP)-induced sepsis is characterized by increased levels of myeloperoxidase (MPO) activity, morphological changes in liver and pro-inflammatory cytokines and chemokines in the liver and lung. SiRNA treatment attenuated inflammation in the liver and lungs of mice following CLP-induced sepsis. Liver MPO activity increased in CLP-induced sepsis and treatment with siRNA significantly reduced this. Similarly, lung MPO activity increased following induction of sepsis with CLP while siRNA treatment significantly reduced MPO activity. Liver and lung cytokine and chemokine levels in CLP-induced sepsis reduced following treatment with siRNA. These findings show a crucial pro-inflammatory role for H2S synthesized by CSE in macrophages in sepsis and suggest CSE gene silencing with siRNA as a potential therapeutic approach for this condition.
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Objective To investigate the relationsheep between colonic H2 S‐producing enzyme cystathionine‐γ‐lyase(CSE) and colonic motility in a rat model of diabetes mellitus(DM) .Methods To obtained diabetic rat models ,all rats were injected intra‐peritoneally (ip) 1% streptozotocin (STZ ,65 mg/kg) ,type 1 diabetes model was established ,nornmol conditions were observed pe‐riodically .Ten days after treatment ,production of colonic longitudinal smooth muscle strips ,organ bath recordings were used to test the colonic motility ;immunofluorescence and double immunofluorescence lableling method were performed to detect the distribution of CSE;Western blot were performed on rat colonic samples devoid of mucosa and submucosa to detect the expression of CSE .Re‐sults DM rats decreased the colonic motility(P0 .05);DM rats increased the expression of CSE in the colon devoid of mucosa and sub‐mucosa(P<0 .05) .Conclusion The decrease of colonic motility in diabetic rat may be associated with the increased production of H2 S‐producing enzyme CSE .
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AIM:To investigate the effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced injury of the mouse podocyte cell line MPC5.METHODS: The cultured MPC5 cells were randomly divided into 4 groups: HG group, normal glucose (NG) group, NG+DL-propargylglycine (PPG) group, and HG+NaHS group.After treated for a certain time, the cells were collected for further detection .The expression of zonula occludens-2 (ZO-2), nephrin,β-cate-nin and cystathionine γ-lyase ( CSE) was determined by Western blotting .RESULTS:High glucose significantly reduced the expression of nephrin, ZO-2 and CSE (P<0.05), while the level of β-catenin was elevated obviously (P<0.05), all in a time-dependent manner.NG+PPG inhibited the levels of ZO-2 and nephrin significantly (P<0.05), and increased the level of β-catenin (P<0.05), all in a PPG concentration-dependent manner.HG+NaHS induced a more significant increase in the levels of ZO-2 and nephrin as compared with HG group (P<0.01), whereas a severe reduction of β-cate-nin in HG+NaHS group was observed as compared with HG group .Compared with NG group , the expression of ZO-2 and nephrin was decreased obviously , and the level of β-catenin was increased in HG +NaHS group.CONCLUSION:Down-regulation of CSE contributes to hyperglycemia-induced podocyte injury .Exogenous H 2 S protects against hyperglycemia-in-duced podocyte injury , possibly through up-regulation of ZO-2 and subsequent suppression of Wnt/β-catenin pathway .
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Aims To observe the changes of endoge-nous hydrogen sulfide/cystathionine-γ-lyase (H2 S/CSE)system and to study the effects of H2 S on cardiac function,H2 S/CSE and myocardial infarct volumes in acute myocardial ischemic injury in isolated hearts of rats.Methods The myocardial ischemic injury model was established by the ligation of coronary artery.The hemodynamic parameters,such as the left ventricular developed pressure (LVDP),±dp/dtmax and coronary arterial flow(CF),were respectively recorded to evalu-ate the cardiac function.The content of H2 S and the activity of CSE in cardiac tissue were detected respec-tively at each time point after ischemia.Infarct vol-umes in isolated rat hearts were determined by dual staining with Evans-blue and TTC.Results (1 )Com-pared with those of the sham group,LVDP,±dp/dtmax and CF were significantly decreased at 30 min,1,2,3, 4 h after ischemia(P<0.01),there were no statistical-ly significant differences in the content of H2 S and the activity of CSE in cardiac tissue at 3 0 min after ische-mia.But during the periods from 1 h to 4h after ische-mia,the content of H2 S and the activity of CSE in car-diac tissue were significantly decreased,the infarct volumes were greatly increased compared with those of the sham control group (P<0.05 or P<0.01 ).(2) Compared with those of the ischemia 4h group,LVDP, ±dp/dtmax and CF were significantly increased in the NaHS low,middle and high dose groups (P<0.05 or P<0.01),the content of H2S and the activity of CSE in cardiac tissue were significantly increased in the NaHS low,middle and high dose groups(P<0.05 or P<0.01 ),and the infarct volumes were significantly decreased in NaHS middle and high dose groups(P<0.01 ).Conclusion H2S and CSE are involved in myocardial ischemic injury in isolated hearts of rats. Administration of NaHS could enhance the activity of CSE,increase the content of H2 S,and reduce infarct volumes.It could be suggested that H2 S has cardiopro-tective effects on acute myocardial ischemic injury.
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Endogenous hydrogen sulfide is the third gaseous signal molecule after nitric oxide and carbon monoxide; it is synthesized through cystathionine- β-synthase and cystathionine-γ-cleavage pathway. Hydrogen sulfide can directly react with the KATP channel, reduce extracellular Ca 2+ influx, and work together with testosterone, nitric oxide, carbon monoxide, relaxing corpus cavernosum smooth muscle and improving diastolic function of blood vessels. It can also inhibit vascular smooth muscle cell proliferation, promote apoptosis, and improve vascular remodeling. Therefore, in-depth study of H2 S in the regulating penile erection can provide a new way for the treatment of erectile dysfunction.