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Apoptosis of bone marrow mesenchymal stem cells caused by homocysteine via activating JNK signal.

Cai B, Li X, Wang Y, Liu Y, Yang F, Chen H, Yin K, Tan X, Zhu J, Pan Z, Wang B, Lu Y - PLoS ONE (2013)

Bottom Line: Apoptosis of BMSCs in response to various pathological stimuli leads to the attenuation of healing ability of BMSCs.Furthermore, apoptotic appearance and mitochondrial membrane potential depolarization in homocysteine-treated BMSCs was significantly reversed by JNK inhibitor but not p38 MAPK and ERK inhibitors.Homocysteine treatment caused a significant reduction of BMSCs-secreted VEGF and IGF-1 in the culture medium.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Harbin Medical University, Harbin, Heilongjiang Province, China.

ABSTRACT
Bone marrow mesenchymal stem cells (BMSCs) are capable of homing to and repair damaged myocardial tissues. Apoptosis of BMSCs in response to various pathological stimuli leads to the attenuation of healing ability of BMSCs. Plenty of evidence has shown that elevated homocysteine level is a novel independent risk factor of cardiovascular diseases. The present study was aimed to investigate whether homocysteine may induce apoptosis of BMSCs and its underlying mechanisms. Here we uncovered that homocysteine significantly inhibited the cellular viability of BMSCs. Furthermore, TUNEL, AO/EB, Hoechst 333342 and Live/Death staining demonstrated the apoptotic morphological appearance of BMSCs after homocysteine treatment. A distinct increase of ROS level was also observed in homocysteine-treated BMSCs. The blockage of ROS by DMTU and NAC prevented the apoptosis of BMSCs induced by homocysteine, indicating ROS was involved in the apoptosis of BMSCs. Moreover, homocysteine also caused the depolarization of mitochondrial membrane potential of BMSCs. Furthermore, apoptotic appearance and mitochondrial membrane potential depolarization in homocysteine-treated BMSCs was significantly reversed by JNK inhibitor but not p38 MAPK and ERK inhibitors. Western blot also confirmed that p-JNK was significantly activated after exposing BMSCs to homocysteine. Homocysteine treatment caused a significant reduction of BMSCs-secreted VEGF and IGF-1 in the culture medium. Collectively, elevated homocysteine induced the apoptosis of BMSCs via ROS-induced the activation of JNK signal, which provides more insight into the molecular mechanisms of hyperhomocysteinemia-related cardiovascular diseases.

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Related in: MedlinePlus

Effects of difference concentrations of homocysteine on the morphological appearance and cellular viability of BMSCs.(a) The morphology of cultured BMSCs was observed after treatment with homocysteine 30, 100, 300 and 1000 µM for 24 h. Homocysteine caused aberrant morphological appearance of BMSCs. (b) Homocysteine significantly decreased the cellular viability of BMSCs in a concentration-dependent manner. * p<0.05 versus Control.
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pone-0063561-g001: Effects of difference concentrations of homocysteine on the morphological appearance and cellular viability of BMSCs.(a) The morphology of cultured BMSCs was observed after treatment with homocysteine 30, 100, 300 and 1000 µM for 24 h. Homocysteine caused aberrant morphological appearance of BMSCs. (b) Homocysteine significantly decreased the cellular viability of BMSCs in a concentration-dependent manner. * p<0.05 versus Control.

Mentions: Firstly, we determine if homocysteine can result in the morphological changes of BMSCs. As shown in Figure 1a, exposure of BMSCs to homocysteine 100, 300 and 1000 µM for 24 h caused obvious cellular morphological changes such as cellular shrinkage. Then, the influence of homocysteine on the cellular viability of BMSCs was assessed by MTT assay. As illuminated in Figure 1b, pretreatment with homocysteine 100, 300 and 1000 µM for 24 h exerted remarkably inhibitory effects on the cellular viability of BMSCs (p<0.05). The cellular viability of BMSCs were significantly decreased by homocysteine 100, 300 and 1000 µM to 85.59±4.69%, 82.82±4.08% and 69.27±9.97 after treatment for 24 h, respectively, but it was not altered by homocysteine 30 µM after treatment for 24 h (Figure 1b). Though the cellular viability of BMSCs was decreased by homocysteine, MTT can not represent the apoptosis of BMSCs induced by homocysteine. Thus, in order to confirm that homocysteine causes BMSCs apoptosis, AO/EB, Hoechest33342 and Live/Death staining were employed in this study.


Apoptosis of bone marrow mesenchymal stem cells caused by homocysteine via activating JNK signal.

Cai B, Li X, Wang Y, Liu Y, Yang F, Chen H, Yin K, Tan X, Zhu J, Pan Z, Wang B, Lu Y - PLoS ONE (2013)

Effects of difference concentrations of homocysteine on the morphological appearance and cellular viability of BMSCs.(a) The morphology of cultured BMSCs was observed after treatment with homocysteine 30, 100, 300 and 1000 µM for 24 h. Homocysteine caused aberrant morphological appearance of BMSCs. (b) Homocysteine significantly decreased the cellular viability of BMSCs in a concentration-dependent manner. * p<0.05 versus Control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3646804&req=5

pone-0063561-g001: Effects of difference concentrations of homocysteine on the morphological appearance and cellular viability of BMSCs.(a) The morphology of cultured BMSCs was observed after treatment with homocysteine 30, 100, 300 and 1000 µM for 24 h. Homocysteine caused aberrant morphological appearance of BMSCs. (b) Homocysteine significantly decreased the cellular viability of BMSCs in a concentration-dependent manner. * p<0.05 versus Control.
Mentions: Firstly, we determine if homocysteine can result in the morphological changes of BMSCs. As shown in Figure 1a, exposure of BMSCs to homocysteine 100, 300 and 1000 µM for 24 h caused obvious cellular morphological changes such as cellular shrinkage. Then, the influence of homocysteine on the cellular viability of BMSCs was assessed by MTT assay. As illuminated in Figure 1b, pretreatment with homocysteine 100, 300 and 1000 µM for 24 h exerted remarkably inhibitory effects on the cellular viability of BMSCs (p<0.05). The cellular viability of BMSCs were significantly decreased by homocysteine 100, 300 and 1000 µM to 85.59±4.69%, 82.82±4.08% and 69.27±9.97 after treatment for 24 h, respectively, but it was not altered by homocysteine 30 µM after treatment for 24 h (Figure 1b). Though the cellular viability of BMSCs was decreased by homocysteine, MTT can not represent the apoptosis of BMSCs induced by homocysteine. Thus, in order to confirm that homocysteine causes BMSCs apoptosis, AO/EB, Hoechest33342 and Live/Death staining were employed in this study.

Bottom Line: Apoptosis of BMSCs in response to various pathological stimuli leads to the attenuation of healing ability of BMSCs.Furthermore, apoptotic appearance and mitochondrial membrane potential depolarization in homocysteine-treated BMSCs was significantly reversed by JNK inhibitor but not p38 MAPK and ERK inhibitors.Homocysteine treatment caused a significant reduction of BMSCs-secreted VEGF and IGF-1 in the culture medium.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Harbin Medical University, Harbin, Heilongjiang Province, China.

ABSTRACT
Bone marrow mesenchymal stem cells (BMSCs) are capable of homing to and repair damaged myocardial tissues. Apoptosis of BMSCs in response to various pathological stimuli leads to the attenuation of healing ability of BMSCs. Plenty of evidence has shown that elevated homocysteine level is a novel independent risk factor of cardiovascular diseases. The present study was aimed to investigate whether homocysteine may induce apoptosis of BMSCs and its underlying mechanisms. Here we uncovered that homocysteine significantly inhibited the cellular viability of BMSCs. Furthermore, TUNEL, AO/EB, Hoechst 333342 and Live/Death staining demonstrated the apoptotic morphological appearance of BMSCs after homocysteine treatment. A distinct increase of ROS level was also observed in homocysteine-treated BMSCs. The blockage of ROS by DMTU and NAC prevented the apoptosis of BMSCs induced by homocysteine, indicating ROS was involved in the apoptosis of BMSCs. Moreover, homocysteine also caused the depolarization of mitochondrial membrane potential of BMSCs. Furthermore, apoptotic appearance and mitochondrial membrane potential depolarization in homocysteine-treated BMSCs was significantly reversed by JNK inhibitor but not p38 MAPK and ERK inhibitors. Western blot also confirmed that p-JNK was significantly activated after exposing BMSCs to homocysteine. Homocysteine treatment caused a significant reduction of BMSCs-secreted VEGF and IGF-1 in the culture medium. Collectively, elevated homocysteine induced the apoptosis of BMSCs via ROS-induced the activation of JNK signal, which provides more insight into the molecular mechanisms of hyperhomocysteinemia-related cardiovascular diseases.

Show MeSH
Related in: MedlinePlus