Limits...
Asymmetric synthesis and evaluation of danshensu-cysteine conjugates as novel potential anti-apoptotic drug candidates.

Pan LL, Wang J, Jia YL, Zheng HM, Wang Y, Zhu YZ - Int J Mol Sci (2014)

Bottom Line: Our results show that DSC and its two diastereoisomers exert similar protective effects in hydrogen peroxide (H2O2)-induced cellular injury in SH-SY5Y cells, as evidenced by the increase of cell viability, superoxide dismutase (SOD), and reduced glutathione (GSH) activity, and glutathione peroxidase (GPx) expression, and the decrease of cellular morphological changes and nuclear condensation, lactate dehydrogenase (LDH) release, and malondialdehyde (MDA) production.In H2O2-stimulated human umbilical vein endothelial cells (HUVEC), DSC concentration-dependently attenuates H2O2-induced cell death, LDH release, mitochondrial membrane potential collapse, and modulates the expression of apoptosis-related proteins (Bcl-2, Bax, caspase-3, and caspase-9).Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Bioactive Small Molecules, Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China. panlilong@fudan.edu.cn.

ABSTRACT
We have previously reported that the danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) is a potent anti-oxidative and anti-apoptotic agent. Herein, we further design and asymmetrically synthesize two diastereoisomers of DSC and explore their potential bioactivities. Our results show that DSC and its two diastereoisomers exert similar protective effects in hydrogen peroxide (H2O2)-induced cellular injury in SH-SY5Y cells, as evidenced by the increase of cell viability, superoxide dismutase (SOD), and reduced glutathione (GSH) activity, and glutathione peroxidase (GPx) expression, and the decrease of cellular morphological changes and nuclear condensation, lactate dehydrogenase (LDH) release, and malondialdehyde (MDA) production. In H2O2-stimulated human umbilical vein endothelial cells (HUVEC), DSC concentration-dependently attenuates H2O2-induced cell death, LDH release, mitochondrial membrane potential collapse, and modulates the expression of apoptosis-related proteins (Bcl-2, Bax, caspase-3, and caspase-9). Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

Show MeSH

Related in: MedlinePlus

The danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) attenuated H2O2-induced cell damage in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the cell viability (A) and LDH (B) release were measured, respectively. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, * p < 0.05 compared with H2O2-stimulated cells. Data were from at least three independent experiments, each performed in duplicate.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4307265&req=5

ijms-16-00628-f005: The danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) attenuated H2O2-induced cell damage in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the cell viability (A) and LDH (B) release were measured, respectively. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, * p < 0.05 compared with H2O2-stimulated cells. Data were from at least three independent experiments, each performed in duplicate.

Mentions: Oxidative stress is one of the mechanisms that cause widespread endothelial dysfunction in cardiovascular diseases and disorders [15]. To examine the endothelial protective effects of Danshensu-cysteine conjugates on oxidative stress-induced cytotoxicity, HUVEC were conditioned with Danshensu-cysteine conjugates for 4 h prior to H2O2 stimulation. Taking into account the difficulty of chemical synthesis, only DSC was chosen to study the protective effects of Danshensu-cysteine conjugates on H2O2-induced cellular injury. As shown in Figure 5A, H2O2 (200 μM) stimulation for 12 h significantly decreased cell viability, which was attenuated by DSC (5–100 μM) in a concentration-dependent manner. Meanwhile, DSC (5–100 μM) also decreased H2O2-induced LDH release in a concentration dependent manner (Figure 5B). In addition, DSC (5–200 μM) treatment for 24 h did not affect cell viability of HUVEC as checked by MTT assay (data not shown). As a positive control, NAC also markedly reduced H2O2-induced cellular damage in HUVEC (Figure 5A,B).


Asymmetric synthesis and evaluation of danshensu-cysteine conjugates as novel potential anti-apoptotic drug candidates.

Pan LL, Wang J, Jia YL, Zheng HM, Wang Y, Zhu YZ - Int J Mol Sci (2014)

The danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) attenuated H2O2-induced cell damage in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the cell viability (A) and LDH (B) release were measured, respectively. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, * p < 0.05 compared with H2O2-stimulated cells. Data were from at least three independent experiments, each performed in duplicate.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307265&req=5

ijms-16-00628-f005: The danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) attenuated H2O2-induced cell damage in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the cell viability (A) and LDH (B) release were measured, respectively. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, * p < 0.05 compared with H2O2-stimulated cells. Data were from at least three independent experiments, each performed in duplicate.
Mentions: Oxidative stress is one of the mechanisms that cause widespread endothelial dysfunction in cardiovascular diseases and disorders [15]. To examine the endothelial protective effects of Danshensu-cysteine conjugates on oxidative stress-induced cytotoxicity, HUVEC were conditioned with Danshensu-cysteine conjugates for 4 h prior to H2O2 stimulation. Taking into account the difficulty of chemical synthesis, only DSC was chosen to study the protective effects of Danshensu-cysteine conjugates on H2O2-induced cellular injury. As shown in Figure 5A, H2O2 (200 μM) stimulation for 12 h significantly decreased cell viability, which was attenuated by DSC (5–100 μM) in a concentration-dependent manner. Meanwhile, DSC (5–100 μM) also decreased H2O2-induced LDH release in a concentration dependent manner (Figure 5B). In addition, DSC (5–200 μM) treatment for 24 h did not affect cell viability of HUVEC as checked by MTT assay (data not shown). As a positive control, NAC also markedly reduced H2O2-induced cellular damage in HUVEC (Figure 5A,B).

Bottom Line: Our results show that DSC and its two diastereoisomers exert similar protective effects in hydrogen peroxide (H2O2)-induced cellular injury in SH-SY5Y cells, as evidenced by the increase of cell viability, superoxide dismutase (SOD), and reduced glutathione (GSH) activity, and glutathione peroxidase (GPx) expression, and the decrease of cellular morphological changes and nuclear condensation, lactate dehydrogenase (LDH) release, and malondialdehyde (MDA) production.In H2O2-stimulated human umbilical vein endothelial cells (HUVEC), DSC concentration-dependently attenuates H2O2-induced cell death, LDH release, mitochondrial membrane potential collapse, and modulates the expression of apoptosis-related proteins (Bcl-2, Bax, caspase-3, and caspase-9).Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Bioactive Small Molecules, Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China. panlilong@fudan.edu.cn.

ABSTRACT
We have previously reported that the danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) is a potent anti-oxidative and anti-apoptotic agent. Herein, we further design and asymmetrically synthesize two diastereoisomers of DSC and explore their potential bioactivities. Our results show that DSC and its two diastereoisomers exert similar protective effects in hydrogen peroxide (H2O2)-induced cellular injury in SH-SY5Y cells, as evidenced by the increase of cell viability, superoxide dismutase (SOD), and reduced glutathione (GSH) activity, and glutathione peroxidase (GPx) expression, and the decrease of cellular morphological changes and nuclear condensation, lactate dehydrogenase (LDH) release, and malondialdehyde (MDA) production. In H2O2-stimulated human umbilical vein endothelial cells (HUVEC), DSC concentration-dependently attenuates H2O2-induced cell death, LDH release, mitochondrial membrane potential collapse, and modulates the expression of apoptosis-related proteins (Bcl-2, Bax, caspase-3, and caspase-9). Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

Show MeSH
Related in: MedlinePlus