The effects of H2 S and its synthetase inhibitor on the autophagy in acute pancreatitis mice / 中华胰腺病杂志

Objective To investigate the effect of H 2 S and its synthetase inhibitor propargylglycine ( PAG) on the autophagic function in caerulein-induced acute pancreatitis ( AP) mice.Methods A total of 60 male BALB/c mice were randomly divided into control , AP, NaHS and PAG group using random number method.AP was induced in mice via hourly intraperitoneal injection of caerulein (50 μg/kg) continuously for 6 hours.NaHS and PAG group received NaHS (10 mg/kg) or PAG (50 mg/kg) 1 h before the AP induction . A equal volume of normal saline solution was injected in control group and AP group .All the mice were killed at 12 h after the first caerulein injection and blood sample was collected for the detection of serum amylase and lipase content.Deproteinization spectrometry was used to detect serum H 2 S content, and pancreatic tissue was pathological examined and scored . Real-time PCR detected mRNA expression of CSE , and the protein expression of LC3-Ⅱ/LC3 Ⅰand p62 was measured using Western blot .Results Serum amylase, lipase, H2S, CSE mRNA, LC3Ⅱ/LC3Ⅰand p62 were (2 700 ±100)U/L, (70 ±20)U/L,(22.9 ±1.7)mmol/L, 1.0 ±0.1,0.419 ±0.080, 0.227 ±0.140 in control group; (17 290 ±500)U/L,(520 ±40)U/L, (31.3 ± 3.0)mmol/L, 5.4 ±0.4, 1.184 ±0.120, 1.985 ±0.210 in AP group; (27 784 ±1 200)U/L, (900 ± 80)U/L,(38.6 ±3.3)mmol/L, 6.9 ±0.9,1.600 ±0.210, 4.229 ±0.050 in NaHS group; (13 750 ± 2 000)U/L,(370 ±20)U/L, (24.5 ±2.1)mmol/L, 4.2 ±0.5, 0.745 ±0.130, 1.203 ±0.080 in PAG group.All those biomarkers detected above in AP group significantly increased compared with control group , which were much lower than those in NaHS group , but higher than those in PAG group , and the differences were statistically significant (all P<0.05).Pancreatic histological damage in NaHS group was more severe than that in AP group , which in PAG group was less severe than that in AP group .Conclusions PAG could greatly decrease serum amylase and lipase level , and reduce the damage on autophagy and the severity of AP .

The protective role of hydrogen sulfide in a mouse model of viral myocarditis induced by Coxsachie-virus B3 / 中华微生物学和免疫学杂志

Objective To investigate the protective role of hydrogen sulfide and the expression of cystathionine gamma-lyase/hydrogen sulfide pathway in a mouse model of myocarditis induced by Coxsachie -virus B3(CVB3).Methods A total of 110 five-week-old BALB/c male mice were randomly divided into four groups:the control group, viral myocarditis group, sodium bisulfide (NaHS) group (50 μmol/kg) and DL-propargylglycine (PAG) group (40 mg/kg).The experimental model of viral myocarditis was induced by intraperitoneal injection of CVB 3.Then the four groups were respectively administered with PBS , PBS, NaHS and PAG from day 1 to day 10 after infection.Blood and heart specimens were harvested from 10 mice of each group on day 4 and day 10 for evaluation of myocardial edema .The pathological changes in heart tis-sues were observed through hematoxylin-eosin staining.Levels of H2 S, IL-6 and TNF-αwere measured by ELISA.The expressions of CSE and CVB 3 at mRNA level were determined by quantitative real time PCR ( qRT -PCR ) analysis and the expression of CSE at protein level was detected by Western blot .Results Compared with the control group , the levels of H2 S and the expressions of CSE at mRNA and protein levels were down-regulated in mice with CVB 3-induced myocarditis .With the treatment of NaHS , the levels of H 2 S in serum and tissue were both up-regulated , and the histopathological damage was alleviated .However , PAG as an irreversible CSE inhibitor inhibited the expressions of H 2 S and CSE and aggravated myocardial injury , inflammatory cells infiltration and interstitial edema .Moreover , the RT-PCR analysis also showed that the expression of CVB3 at mRNA level was inhibited by NaHS but enhanced by PAG .Conclusion The expres-sion of CSE/H2 S pathway is down-regulated in mice with CVB 3-induced viral myocarditis .PAG could pro-mote virus propagation and exacerbate the disease through inhibiting the production of endogenous H 2 S, while NaHS as a H2 S donor has a protective effect on infected myocardium by suppressing virus replication at an early stage .

Effect of endogenous hydrogen sulfide inhibition on structural and functional renal disturbances induced by gentamicin

Animal models of gentamicin nephrotoxicity present acute tubular necrosis associated with inflammation, which can contribute to intensify the renal damage. Hydrogen sulfide (H2S) is a signaling molecule involved in inflammation. We evaluated the effect of DL-propargylglycine (PAG), an inhibitor of endogenous H2S formation, on the renal damage induced by gentamicin. Male Wistar rats (N = 8) were injected with 40 mg/kg gentamicin (im) twice a day for 9 days, some of them also received PAG (N = 8, 10 mg·kg-1·day-1, ip). Control rats (N = 6) were treated with saline or PAG only (N = 4). Twenty-four-hour urine samples were collected one day after the end of these treatments, blood samples were collected, the animals were sacrificed, and the kidneys were removed for quantification of H2S formation and histological and immunohistochemical studies. Gentamicin-treated rats presented higher sodium and potassium fractional excretion, increased plasma creatinine [4.06 (3.00; 5.87) mg percent] and urea levels, a greater number of macrophages/monocytes, and a higher score for tubular interstitial lesions [3.50 (3.00; 4.00)] in the renal cortex. These changes were associated with increased H2S formation in the kidneys from gentamicin-treated rats (230.60 ± 38.62 µg·mg protein-1·h-1) compared to control (21.12 ± 1.63) and PAG (11.44 ± 3.08). Treatment with PAG reduced this increase (171.60 ± 18.34), the disturbances in plasma creatinine levels [2.20 (1.92; 4.60) mg percent], macrophage infiltration, and score for tubular interstitial lesions [2.00 (2.00; 3.00)]. However, PAG did not interfere with the increase in fractional sodium excretion provoked by gentamicin. The protective effect of PAG on gentamicin nephrotoxicity was related, at least in part, to decreased H2S formation.

Impact of endogenous hydrogen sulfide on pulmonary vascular structural remodeling and vasoactive peptides in rats with high pulmonary blood flow / 中国药理学通报

Aim To explore the impact of endogenous hydrogen sulfide on pulmonary vascular structural remodeling and vasoactive peptides in rats with high pulmonary blood flow. Methods Thirty-two male SD rats, weighing 120~140 g, were randomly divided into shunt group (n=8), shunt+PPG (propargylglycine)group (n=8), control group (n=8) and control+PPG (n=8). Rats in shunt group and shunt+PPG group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. Rats in shunt+PPG group and control+PPG group were intraperitoneally injected with an inhibitor of endogenous H2S generation enzyme-PPG at a dose of 37.5 mg?kg-1 each day. After 4 weeks of experiment, the morphologic changes including micro-and ultra-structural changes of pulmonary arteries of rats were observed under optical microscope and electro-microscope, respectively. H2S concentration in lung tissue was evaluated by sensitive modified sulfide electrode method. Endothelin-1 (ET-1), atrial natriuretic peptide (ANP), calcitonin gene related peptide (CGRP) and adrenomedullin (ADM) were calculated by radioimmunoussay kit. Results After 4 weeks of shunt, lung tissue H2S level increased significantly (P