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Hydrogen sulfide acts as a mediator of inflammation in acute pancreatitis: in vitro studies using isolated mouse pancreatic acinar cells.

Tamizhselvi R, Moore PK, Bhatia M - J. Cell. Mol. Med. (2007 Mar-Apr)

Bottom Line: Caerulein increased the levels of H(2)S and CSE mRNA expression while CBS mRNA expression was decreased.Inhibition of endogenous production of H(2)S by PAG significantly suppressed SP concentration, PPT-A expression and NK1-R expression in the acini.To determine whether H(2)S itself provoked inflammation in acinar cells, the cells were treated with H(2)S donor drug, sodium hydrosulphide (NaHS), (10, 50 and 100 muM), that resulted in a significant increase in SP concentration and expression of PPT-A and NK1-R in acinar cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, National University of Singapore, Yong Loo Lin School of Medicine, Centre for the Sciences, 28 Medical Drive, Singapore 117456.

ABSTRACT
Hydrogen sulphide (H(2)S) is synthesized from L-cysteine via the action of cystathionine-gamma-lyase (CSE) and cystathionine-beta-synthase (CBS). We have earlier shown that H(2)S acts as a mediator of inflammation. However the mechanism remains unclear. In this study, we investigated the presence of H(2)S and the expression of H(2)S synthesizing enzymes, CSE and CBS, in isolated mouse pancreatic acini. Pancreatic acinar cells from mice were incubated with or without caerulein (10(-7) M for 30 and 60 min). Caerulein increased the levels of H(2)S and CSE mRNA expression while CBS mRNA expression was decreased. In addition, cells pre-treated with DL-propargylglycine (PAG, 3 mM), a CSE inhibitor, reduced the formation of H(2)S in caerulein treated cells, suggesting that CSE may be the main enzyme involved in H(2)S formation in mouse acinar cells. Furthermore, substance P (SP) concentration in the acini and expression of SP gene (preprotachykinin-A, PPT-A) and neurokinin-1 receptor (NK-1R), the primary receptor for SP, are increased in secretagogue caerulein-treated acinar cells. Inhibition of endogenous production of H(2)S by PAG significantly suppressed SP concentration, PPT-A expression and NK1-R expression in the acini. To determine whether H(2)S itself provoked inflammation in acinar cells, the cells were treated with H(2)S donor drug, sodium hydrosulphide (NaHS), (10, 50 and 100 muM), that resulted in a significant increase in SP concentration and expression of PPT-A and NK1-R in acinar cells. These results suggest that the pro-inflammatory effect of H(2)S may be mediated by SP-NK-1R related pathway in mouse pancreatic acinar cells.

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Cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS) mRNA expression in control and caerulein-treated mouse pancreatic acinar cells. (A) RT-PCR of CSE in control and caerulein (10−7 M for 30 and 60 min) treated acinar cells. The graphs represent the optical density of the bands of CSE generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (60 min). (B) RT-PCR of CBS in control and caerulein treated acinar cells. The graphs represent the optical density of the bands of CBS generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (0 hr).
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fig02: Cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS) mRNA expression in control and caerulein-treated mouse pancreatic acinar cells. (A) RT-PCR of CSE in control and caerulein (10−7 M for 30 and 60 min) treated acinar cells. The graphs represent the optical density of the bands of CSE generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (60 min). (B) RT-PCR of CBS in control and caerulein treated acinar cells. The graphs represent the optical density of the bands of CBS generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (0 hr).

Mentions: Pancreatic acinar cells were found to express the gene for the H2S forming enzyme CSE and CBS, as evidenced by RT-PCR extracted from these cells (Fig. 2A and B). In addition, incubation of acinar cell homogenates with L-cysteine resulted in the formation of H2S, showing the presence of H2S synthesizing enzyme activity in the cells (Fig. 3A).


Hydrogen sulfide acts as a mediator of inflammation in acute pancreatitis: in vitro studies using isolated mouse pancreatic acinar cells.

Tamizhselvi R, Moore PK, Bhatia M - J. Cell. Mol. Med. (2007 Mar-Apr)

Cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS) mRNA expression in control and caerulein-treated mouse pancreatic acinar cells. (A) RT-PCR of CSE in control and caerulein (10−7 M for 30 and 60 min) treated acinar cells. The graphs represent the optical density of the bands of CSE generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (60 min). (B) RT-PCR of CBS in control and caerulein treated acinar cells. The graphs represent the optical density of the bands of CBS generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (0 hr).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822830&req=5

fig02: Cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS) mRNA expression in control and caerulein-treated mouse pancreatic acinar cells. (A) RT-PCR of CSE in control and caerulein (10−7 M for 30 and 60 min) treated acinar cells. The graphs represent the optical density of the bands of CSE generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (60 min). (B) RT-PCR of CBS in control and caerulein treated acinar cells. The graphs represent the optical density of the bands of CBS generated from six independent experiments that were normalized with the expression of 18S. ***P < 0.001 when caerulein treated cells (60 min) were compared with control cells (0 hr).
Mentions: Pancreatic acinar cells were found to express the gene for the H2S forming enzyme CSE and CBS, as evidenced by RT-PCR extracted from these cells (Fig. 2A and B). In addition, incubation of acinar cell homogenates with L-cysteine resulted in the formation of H2S, showing the presence of H2S synthesizing enzyme activity in the cells (Fig. 3A).

Bottom Line: Caerulein increased the levels of H(2)S and CSE mRNA expression while CBS mRNA expression was decreased.Inhibition of endogenous production of H(2)S by PAG significantly suppressed SP concentration, PPT-A expression and NK1-R expression in the acini.To determine whether H(2)S itself provoked inflammation in acinar cells, the cells were treated with H(2)S donor drug, sodium hydrosulphide (NaHS), (10, 50 and 100 muM), that resulted in a significant increase in SP concentration and expression of PPT-A and NK1-R in acinar cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, National University of Singapore, Yong Loo Lin School of Medicine, Centre for the Sciences, 28 Medical Drive, Singapore 117456.

ABSTRACT
Hydrogen sulphide (H(2)S) is synthesized from L-cysteine via the action of cystathionine-gamma-lyase (CSE) and cystathionine-beta-synthase (CBS). We have earlier shown that H(2)S acts as a mediator of inflammation. However the mechanism remains unclear. In this study, we investigated the presence of H(2)S and the expression of H(2)S synthesizing enzymes, CSE and CBS, in isolated mouse pancreatic acini. Pancreatic acinar cells from mice were incubated with or without caerulein (10(-7) M for 30 and 60 min). Caerulein increased the levels of H(2)S and CSE mRNA expression while CBS mRNA expression was decreased. In addition, cells pre-treated with DL-propargylglycine (PAG, 3 mM), a CSE inhibitor, reduced the formation of H(2)S in caerulein treated cells, suggesting that CSE may be the main enzyme involved in H(2)S formation in mouse acinar cells. Furthermore, substance P (SP) concentration in the acini and expression of SP gene (preprotachykinin-A, PPT-A) and neurokinin-1 receptor (NK-1R), the primary receptor for SP, are increased in secretagogue caerulein-treated acinar cells. Inhibition of endogenous production of H(2)S by PAG significantly suppressed SP concentration, PPT-A expression and NK1-R expression in the acini. To determine whether H(2)S itself provoked inflammation in acinar cells, the cells were treated with H(2)S donor drug, sodium hydrosulphide (NaHS), (10, 50 and 100 muM), that resulted in a significant increase in SP concentration and expression of PPT-A and NK1-R in acinar cells. These results suggest that the pro-inflammatory effect of H(2)S may be mediated by SP-NK-1R related pathway in mouse pancreatic acinar cells.

Show MeSH