Treadmill exercise exerts a synergistic effect with bone marrow mesenchymal stem cell-derived exosomes on neuronal apoptosis and synaptic-axonal remodeling.

Treadmill exercise and mesenchymal stem cell transplantation are both practical and effective methods for the treatment of cerebral ischemia. However, whether there is a synergistic effect between the two remains unclear. In this study, we established rat models of ischemia/reperfusion injury by occlusion of the middle cerebral artery for 2 hours and reperfusion for 24 hours. Rat models were perfused with bone marrow mesenchymal stem cell-derived exosomes (MSC-exos) via the tail vein and underwent 14 successive days of treadmill exercise. Neurological assessment, histopathology, and immunohistochemistry results revealed decreased neuronal apoptosis and cerebral infarct volume, evident synaptic formation and axonal regeneration, and remarkably recovered neurological function in rats subjected to treadmill exercise and MSC-exos treatment. These effects were superior to those in rats subjected to treadmill exercise or MSC-exos treatment alone. Mechanistically, further investigation revealed that the activation of JNK1/c-Jun signaling pathways regulated neuronal apoptosis and synaptic-axonal remodeling. These findings suggest that treadmill exercise may exhibit a synergistic effect with MSC-exos treatment, which may be related to activation of the JNK1/c-Jun signaling pathway. This study provides novel theoretical evidence for the clinical application of treadmill exercise combined with MSC-exos treatment for ischemic cerebrovascular disease.

[Effects of adipose-derived stem cells transplantation on the neuronal apoptosis and the expression of Bcl-2 and caspase-12 in the brain post focal cerebral ischemia in rats].

AIM: To investigate the effects of adipose-derived stem cells (ADSCs) transplantation on neuronal apoptosis in the brain after focal cerebral ischemia in rats. METHODS: 72 male adult Sprague-Dawley rats were randomly divided into 4 groups: Sham-operated group , Middle cerebral artery occlusion (MCAO) group, Vehicle group and ADSC-treated group (n=18). MCAO model was established with the modified Longa's method. One day after right MCAO, 30 µL of cell suspension containing 1×10(6); cells were injected into the lateral ventricle of ADSC-treated group and the same dose of PBS was given to the vehicle group. At 4 d, 7 d and 14 d after MCAO, the apoptosis of neuron was detected by terminal deoxynucleotidyl transferase-mediated DNA nick-end labeling (TUNEL) method and the expression of Bcl-2 and caspase-12 in ischemic region was detected by immunohistochemistry and RT-PCR. RESULTS: TUNEL-positive cells in ischemic region of ADSC-treated group were less than that in MCAO group and Vehicle group at 4 d, 7 d and 14 d post MCAO (P<0.05). Compared with MCAO group and Vehicle group, the expression of Bcl-2 significantly up-regulated while caspase-12 expression significantly decreased in ADSC-treated group at any time point post MCAO (P<0.05). CONCLUSION: The transplantation of ADSCs can reduce neuronal apoptosis of rats with cerebral ischemic injury partly by promoting the expression of Bcl-2 which participates in apoptotic signals after mitochondrial damage and inhibiting the expression of caspase-12 which mediates endoplasmic reticulum (ER) stress-induced apoptosis.

[The effects of adipose-derived stem cell transplantation on the expression of IL-10 and TNF-alpha after cerebral ischaemia in rats].

AIM: To explore the effects of adipose-derived stem cell (ADSC) transplantation on the expression of IL-10 amd TNF-alpha after cerebral ischaemia in Middle cerebral artery occlusion (MCAO) rats. METHODS: 72 male adult Sprague-Dawley rats were randomly divided into 4 groups: sham group, MCAO group, Vehicle group and ADSC group (n=18). Rat's cerebral ischemia model was established by MCAO with Longa's method. ADSC were labeled by DAPI before transplantation. One day after MCAO, 30 microL of cell suspension containing 1 x 10(6); ADSCs were injected into the lateral ventricle of ADSC group and the same dose of PBS was given to the Vehicle group. At day 4, day 7 and day 14 after MCAO, the rats were decapitated to detect the expression of IL-10 and TNF-alpha in ischaemic rat's brain by ELISA, immunohistochemistry and RT-PCR. RESULTS: Compared with sham group, the expression of IL-10 and TNF-alpha significantly up-regulated at 4 d, 7 d of MCAO group(P<0.05). There was no statistical difference of IL-10 and TNF-alpha expression between MCAO and vehicle group ant any time point(P>0.05). Compared with Vehicle group, the expression of IL-10 significantly up-regulated while TNF-alpha expression significantly decreased of ADSC-treated group at any timepoint post MCAO(P<0.05). CONCLUSION: The transplantation of ADSC could up-regulate the expression of IL-10 and down-regulate the expression of TNF-alpha in MCAO rat's brain, which might contribute to its protective role upon cerebral ischaemia.

[Effects of adipose-derived stem cell transplantation on the angiogenesis and the expression of bFGF and VEGF in the brain post focal cerebral ischemia in rats].

AIM: To investigate the effects of adipose-derived stem cell (ADSC) transplantation on the angiogenesis in the brain post focal cerebral ischemia in rats. METHODS: 72 male adult Sprague-Dawley rats were randomly divided into 4 groups: sham-operated group, middle cerebral artery occlusion (MCAO) group, vehicle group and MCAO+ADSC-treated group (n=18). A permenant focal cerebral ischemia model was established with the modified Longa's method. ADSC were labeled by DAPI before transplantation. One day after right MCAO, 30 muL of cell suspension containing 1x10(6) cells were injected into the lateral ventricle of MCAO+ADSC-treated group and the same dose of PBS was given to the vehicle group. On D4, D7 and D14 after MCAO, the rats were killed to detect the regeneration of microvessel and the expression of bFGF and VEGF in ischemic region by immunohistochemistry and RT-PCR. RESULTS: A lot of microvessel proliferate in the injured cortex reached peak in 2 weeks. The microvessel density in the brain tissues of rats treated with ADSC was higher than that in MCAO group and vehicle group (P<0.01). The expression of bFGF and VEGF in the brain tissues of MCAO+ADSC-treated group was higher than that in MCAO group and vehicle group on D4, D7 and D14 post MCAO. CONCLUSION: The transplantation of ADSC can promote the revascularization of cerebral ischemia in rats partly by enhancing bFGF and VEGF synthesis in brain.