Limits...
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.

Show MeSH

Related in: MedlinePlus

Effects of homocysteine on intracellular ROS and mitochondrial membrane potential of BMSCs.(a) Intracellular ROS level was measured in BMSCs treated with homocysteine 30, 100 and 300 µM for 24 h by DCFH-DA staining. The ROS level was gradually increased with the increase of homocysteine concentration. (b) Homocysteine induced an obvious depolarization of mitochondrial membrane potential in BMSCs apoptosis by JC-1 staining.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3646804&req=5

pone-0063561-g003: Effects of homocysteine on intracellular ROS and mitochondrial membrane potential of BMSCs.(a) Intracellular ROS level was measured in BMSCs treated with homocysteine 30, 100 and 300 µM for 24 h by DCFH-DA staining. The ROS level was gradually increased with the increase of homocysteine concentration. (b) Homocysteine induced an obvious depolarization of mitochondrial membrane potential in BMSCs apoptosis by JC-1 staining.

Mentions: It is well documented that reactive oxygen species (ROS) is involved in apoptosis of many cell types [24]. Oxidative stresses caused by ROS are shown to initiate or promote apoptosis via oxidizing mitochondrial membrane phospholipids and depolarizing mitochondrial membrane potential which produces more ROS [24], [25]. We therefore investigated the influences of homocysteine on the production of ROS and mitochondrial membrane potential by DCFH-DA staining and JC-1 staining, respectively. As shown in Figure 3a, DCFH-DA staining showed that both the intensity of green inflorescence and the percentage of ROS-positive cells were significantly increased in the presence of homocysteine 300 µM for 24 h. Moreover, treatment of BMSCs with homocysteine for 24 h was able to cause the obvious depolarization of mitochondrial membrane potential (Figure 3b). These indicate that ROS-mediated mitochondrial dysfunction is involved in homocysteine-induced BMSCs apoptosis.


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 homocysteine on intracellular ROS and mitochondrial membrane potential of BMSCs.(a) Intracellular ROS level was measured in BMSCs treated with homocysteine 30, 100 and 300 µM for 24 h by DCFH-DA staining. The ROS level was gradually increased with the increase of homocysteine concentration. (b) Homocysteine induced an obvious depolarization of mitochondrial membrane potential in BMSCs apoptosis by JC-1 staining.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0063561-g003: Effects of homocysteine on intracellular ROS and mitochondrial membrane potential of BMSCs.(a) Intracellular ROS level was measured in BMSCs treated with homocysteine 30, 100 and 300 µM for 24 h by DCFH-DA staining. The ROS level was gradually increased with the increase of homocysteine concentration. (b) Homocysteine induced an obvious depolarization of mitochondrial membrane potential in BMSCs apoptosis by JC-1 staining.
Mentions: It is well documented that reactive oxygen species (ROS) is involved in apoptosis of many cell types [24]. Oxidative stresses caused by ROS are shown to initiate or promote apoptosis via oxidizing mitochondrial membrane phospholipids and depolarizing mitochondrial membrane potential which produces more ROS [24], [25]. We therefore investigated the influences of homocysteine on the production of ROS and mitochondrial membrane potential by DCFH-DA staining and JC-1 staining, respectively. As shown in Figure 3a, DCFH-DA staining showed that both the intensity of green inflorescence and the percentage of ROS-positive cells were significantly increased in the presence of homocysteine 300 µM for 24 h. Moreover, treatment of BMSCs with homocysteine for 24 h was able to cause the obvious depolarization of mitochondrial membrane potential (Figure 3b). These indicate that ROS-mediated mitochondrial dysfunction is involved in homocysteine-induced BMSCs apoptosis.

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