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Urotensin II Protects Cardiomyocytes from Apoptosis Induced by Oxidative Stress through the CSE/H2S Pathway.

Gong H, Chen Z, Zhang X, Li Y, Zhang J, Chen Y, Ding Y, Zhang G, Yang C, Zhu Y, Zou Y - Int J Mol Sci (2015)

Bottom Line: UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2.SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects.In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China. ghui1975@163.com.

ABSTRACT
Plasma urotensin II (UII) has been observed to be raised in patients with acute myocardial infarction; suggesting a possible cardiac protective role for this peptide. However, the molecular mechanism is unclear. Here, we treated cultured cardiomyocytes with H2O2 to induce oxidative stress; observed the effect of UII on H2O2-induced apoptosis and explored potential mechanisms. UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2. SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects. Further analysis revealed that UII increased the production of hydrogen sulfide (H2S) and the level of cystathionine-γ-lyase (CSE) by activating the ERK signaling in H2O2-treated-cardiomyocytes. Si-CSE or ERK inhibitor not only greatly inhibited the increase in CSE level or the phosphorylation of ERK induced by UII but also reversed anti-apoptosis of UII in H2O2-treated-cadiomyocytes. In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress. These results suggest that increased plasma UII level may protect cardiomyocytes at the early-phase of acute myocardial infarction in patients.

No MeSH data available.


Related in: MedlinePlus

UII increases the production of H2S by enhancing the level of CSE in cardiomyocytes exposed to H2O2. (A) H2S production was examined in culture supernatant of cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. 0 h group; #p < 0.05 vs. 12 h group; (B) H2S production was examined in culture medium of cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. si-UT: Cardiomyocytes (CMs) were transfected with siRNA-UT, or siRNA-scramble as control. Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. H2O2 group; ##p < 0.01 vs. H2O2 + UII group; (C) RT-PCR analysis of CBS and CSE mRNA expression in cultured cardiomyocytes. The expression of CBS and CSE mRNAs in liver tissue was detected as positive control; (D) Western blot analysis of CSE level in cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05 vs. 0 h group; #p < 0.05 vs. 12 h group; (E) Western blot analysis of CSE level in cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. CMs: Cardiomyocytes. Si-UT: CMs were transfected with siRNA-UT; Si-scram: CMs were transfected with siRNA-scramble as control. Values are mean ± SEM. *p < 0.05 vs. H2O2 group; #p < 0.05 vs. H2O2 + UII group. All experiments were repeated independently at least three times.
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ijms-16-12482-f002: UII increases the production of H2S by enhancing the level of CSE in cardiomyocytes exposed to H2O2. (A) H2S production was examined in culture supernatant of cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. 0 h group; #p < 0.05 vs. 12 h group; (B) H2S production was examined in culture medium of cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. si-UT: Cardiomyocytes (CMs) were transfected with siRNA-UT, or siRNA-scramble as control. Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. H2O2 group; ##p < 0.01 vs. H2O2 + UII group; (C) RT-PCR analysis of CBS and CSE mRNA expression in cultured cardiomyocytes. The expression of CBS and CSE mRNAs in liver tissue was detected as positive control; (D) Western blot analysis of CSE level in cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05 vs. 0 h group; #p < 0.05 vs. 12 h group; (E) Western blot analysis of CSE level in cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. CMs: Cardiomyocytes. Si-UT: CMs were transfected with siRNA-UT; Si-scram: CMs were transfected with siRNA-scramble as control. Values are mean ± SEM. *p < 0.05 vs. H2O2 group; #p < 0.05 vs. H2O2 + UII group. All experiments were repeated independently at least three times.

Mentions: Recently, H2S has been reported to protect against apoptosis in cardiomyocytes stimulated with isoproterenol [29] or oxidative stress [30,31]. We therefore studied the effect of UII on H2S production in cardiomyocytes. UII treatment sharply increased the production of H2S in cardiomyocytes in time-dependent manner. After 24 h, H2S production increased more than three-fold in UII-treated-cardiomyocytes, when compared with control (Figure 2A). H2O2 has little effect on the production of H2S in cardiomyocytes, but UII-pretreatment greatly increased H2S production, and si-UT partly abolished the effect of UII in H2O2-treated-cardiomyocytes (Figure 2B). Endogenous H2S is usually produced by CBS or CSE in mammalian systems. CSE is the major H2S-producing enzyme in the cardiovascular system [10,12]. Reverse transcription PCR showed that CSE mRNA, not CBS mRNA, was detected in cultured cardiomyocytes (Figure 2C). Western blot analysis revealed that UII significantly upregulated the level of CSE in cardiomyocytes in a time-dependent manner (Figure 2D). At two days after myocardial infarction, UII mRNA level was increased about three-fold compared to that in the sham group, and CSE expression was upregulated accordingly after myocardial infarction (Figure S1B,D). In cultured cardiomyocytes, H2O2 did not affect the CSE level but UII greatly increased the level in the presence of H2O2, and the effect was partly abolished by si-UT (Figure 2E). These data revealed that UII promoted H2S production by upregulating CSE level in H2O2-treated-cardiomyocytes.


Urotensin II Protects Cardiomyocytes from Apoptosis Induced by Oxidative Stress through the CSE/H2S Pathway.

Gong H, Chen Z, Zhang X, Li Y, Zhang J, Chen Y, Ding Y, Zhang G, Yang C, Zhu Y, Zou Y - Int J Mol Sci (2015)

UII increases the production of H2S by enhancing the level of CSE in cardiomyocytes exposed to H2O2. (A) H2S production was examined in culture supernatant of cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. 0 h group; #p < 0.05 vs. 12 h group; (B) H2S production was examined in culture medium of cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. si-UT: Cardiomyocytes (CMs) were transfected with siRNA-UT, or siRNA-scramble as control. Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. H2O2 group; ##p < 0.01 vs. H2O2 + UII group; (C) RT-PCR analysis of CBS and CSE mRNA expression in cultured cardiomyocytes. The expression of CBS and CSE mRNAs in liver tissue was detected as positive control; (D) Western blot analysis of CSE level in cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05 vs. 0 h group; #p < 0.05 vs. 12 h group; (E) Western blot analysis of CSE level in cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. CMs: Cardiomyocytes. Si-UT: CMs were transfected with siRNA-UT; Si-scram: CMs were transfected with siRNA-scramble as control. Values are mean ± SEM. *p < 0.05 vs. H2O2 group; #p < 0.05 vs. H2O2 + UII group. All experiments were repeated independently at least three times.
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Related In: Results  -  Collection

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ijms-16-12482-f002: UII increases the production of H2S by enhancing the level of CSE in cardiomyocytes exposed to H2O2. (A) H2S production was examined in culture supernatant of cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. 0 h group; #p < 0.05 vs. 12 h group; (B) H2S production was examined in culture medium of cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. si-UT: Cardiomyocytes (CMs) were transfected with siRNA-UT, or siRNA-scramble as control. Values are mean ± SEM. *p < 0.05, **p < 0.01 vs. H2O2 group; ##p < 0.01 vs. H2O2 + UII group; (C) RT-PCR analysis of CBS and CSE mRNA expression in cultured cardiomyocytes. The expression of CBS and CSE mRNAs in liver tissue was detected as positive control; (D) Western blot analysis of CSE level in cardiomyocytes treated with UII at different time points (0, 12, 24, 48 h). Values are mean ± SEM. *p < 0.05 vs. 0 h group; #p < 0.05 vs. 12 h group; (E) Western blot analysis of CSE level in cardiomyocytes transfected with si-UT or siRNA-scramble for 48 h, then treated with UII (0.1 μM) or PBS for 30 min and exposed to 100 μM H2O2 for 24 h. CMs: Cardiomyocytes. Si-UT: CMs were transfected with siRNA-UT; Si-scram: CMs were transfected with siRNA-scramble as control. Values are mean ± SEM. *p < 0.05 vs. H2O2 group; #p < 0.05 vs. H2O2 + UII group. All experiments were repeated independently at least three times.
Mentions: Recently, H2S has been reported to protect against apoptosis in cardiomyocytes stimulated with isoproterenol [29] or oxidative stress [30,31]. We therefore studied the effect of UII on H2S production in cardiomyocytes. UII treatment sharply increased the production of H2S in cardiomyocytes in time-dependent manner. After 24 h, H2S production increased more than three-fold in UII-treated-cardiomyocytes, when compared with control (Figure 2A). H2O2 has little effect on the production of H2S in cardiomyocytes, but UII-pretreatment greatly increased H2S production, and si-UT partly abolished the effect of UII in H2O2-treated-cardiomyocytes (Figure 2B). Endogenous H2S is usually produced by CBS or CSE in mammalian systems. CSE is the major H2S-producing enzyme in the cardiovascular system [10,12]. Reverse transcription PCR showed that CSE mRNA, not CBS mRNA, was detected in cultured cardiomyocytes (Figure 2C). Western blot analysis revealed that UII significantly upregulated the level of CSE in cardiomyocytes in a time-dependent manner (Figure 2D). At two days after myocardial infarction, UII mRNA level was increased about three-fold compared to that in the sham group, and CSE expression was upregulated accordingly after myocardial infarction (Figure S1B,D). In cultured cardiomyocytes, H2O2 did not affect the CSE level but UII greatly increased the level in the presence of H2O2, and the effect was partly abolished by si-UT (Figure 2E). These data revealed that UII promoted H2S production by upregulating CSE level in H2O2-treated-cardiomyocytes.

Bottom Line: UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2.SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects.In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China. ghui1975@163.com.

ABSTRACT
Plasma urotensin II (UII) has been observed to be raised in patients with acute myocardial infarction; suggesting a possible cardiac protective role for this peptide. However, the molecular mechanism is unclear. Here, we treated cultured cardiomyocytes with H2O2 to induce oxidative stress; observed the effect of UII on H2O2-induced apoptosis and explored potential mechanisms. UII pretreatment significantly reduced the number of apoptotic cardiomyocytes induced by H2O2; and it partly abolished the increase of pro-apoptotic protein Bax and the decrease of anti-apoptotic protein Bcl-2 in cardiomyocytes induced by H2O2. SiRNA targeted to the urotensin II receptor (UT) greatly inhibited these effects. Further analysis revealed that UII increased the production of hydrogen sulfide (H2S) and the level of cystathionine-γ-lyase (CSE) by activating the ERK signaling in H2O2-treated-cardiomyocytes. Si-CSE or ERK inhibitor not only greatly inhibited the increase in CSE level or the phosphorylation of ERK induced by UII but also reversed anti-apoptosis of UII in H2O2-treated-cadiomyocytes. In conclusion, UII rapidly promoted the phosphorylation of ERK and upregulated CSE level and H2S production, which in turn activated ERK signaling to protect cardiomyocytes from apoptosis under oxidative stress. These results suggest that increased plasma UII level may protect cardiomyocytes at the early-phase of acute myocardial infarction in patients.

No MeSH data available.


Related in: MedlinePlus