Int J Stem Cells.  2022 May;15(2):217-226. 10.15283/ijsc21136.

Effects and Mechanisms of Bone Marrow Mesenchymal Stem Cell Transplantation for Treatment of Ischemic Stroke in Hypertensive Rats

• Liu Y
• Zhao Y
• Min Y
• Guo K
• Chen Y
• Huang Z
• Long C

• KMID: 2529836
• DOI: http://doi.org/10.15283/ijsc21136

Abstract

Background and Objectives
Stroke is the most common cause of human death and functional disability, resulting in more than 5 million deaths worldwide each year. Bone marrow mesenchymal stem cells (BMSCs) are a kind of stem cell that are able to self-renew and differentiate into many types of tissues. Therefore, BMSCs have the potential to replace damaged neurons and promote the reconstruction of nerve conduction pathways and connective tissue. However, it remains unknown whether transplanted BMSCs promote angiogenesis or improve the tissue microenvironment directly or indirectly through paracrine interactions. This study aimed to determine the therapeutic effect of BMSCs on ischemic stroke with hypertension in a rodent model and to explore the possible mechanisms underlying any benefits.
Methods and Results
Middle cerebral artery occlusion was used to establish the experimental stroke model. The area of cerebral infarction, expression of vascular endothelial growth factor (VEGF) and glial cell line-derived neurotrophic factor (GDNF), and increment of astrocyte were measured by TTC staining, western blot, real-time quantitative polymerase chain reaction (RT-qPCR) and immunocytochemistry. The results showed a smaller area of cerebral infarction and improved neurological function scores in animals treated with BMSCs compared to controls. The results of RT-qPCR and western blot assays showed higher expression of VEGF and GDNF in BMSC-treated animals compared with controls. Our study also showed that one round of BMSCs transplantation significantly promoted the proliferation of subventricular zone and cortical cells, especially astrocytes, on the ischemic side following cerebral ischemia.
Conclusions
Above findings support that BMSCs have therapeutic effects for ischemic stroke complicated with hypertension, which may occur via up-regulated expression of VEGF and GDNF and reduction of neuronal apoptosis, thereby promoting the recovery of nerve function.

Keyword

Bone marrow mesenchymal stem cells; Ischemia; Stroke; Hypertensive

Figure

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  Fig. 1 Neurobehavioral scores of rats in each group (*p<0.05 vs Stroke+Vehicle). mNSS test were assessed on day 1, 3, 7, and 14 after surgery respectively. The testing score of BMSC-treated rat was significantly better than the score of vehicle-treated stroke mice. *p<0.05 (mNSS: modified Neurological Severity Scores).

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  Fig. 2 BMSCs transplantation and neuroprotective effect after focal ischemia. (A) 2,3,5-Triphenyltetrazolium chloride (TTC) staining of coronal brain sections from vehicle-treated and BMSCs-treated barrel cortex stroke rat reveals the infarct area (white) in the cortex. (B) Quantified data show that the infarct volume (mean±SD) of BMSC-treated brains was signifi-cantly reduced compared with the vehicle-treated brain (*p<0.05 vs Stro-ke＋Vehicle) (TTC: 2,3,5-Triphenylttra-zolium chloride).

• 

  Fig. 3 Expression of GDNF and VEGF proteins in the peri-infarct cortex. The protein expression levels of GDNF and VEGF in each group were analyzed with Western blot on days 3, 7, and 14 respectively (A). Representative bands of growth associated GDNF and VEGF expression levels (B, C). Quantitative determination of immune proteins of GDNF and VEGF, GDNF and VEGF were quantitatively analyzed on days 3, 7 and 14 in each group. Compared with the sham group and model group, BMSCs therapy significantly enhanced the expression of GDNF and VEGF (*p<0.05, **p<0.01) (VEGF: vascular endothelial growth factor; GDNF: glial cell line-derived neurotrophic factor).

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  Fig. 4 Expression of GDNF and VEGF mRNA in the peri-infarct cortex. The mRNA expression levels of GDNF and VEGF were assessed by RT-PCR at different time periods (A, B). GDNF and VEGF were quantitatively analyzed on days 3, 7 and 14 in each group. Compared with the sham group and model group, BMSCs group significantly increased the expression of GDNF and VEGF (*p<0.05, **p<0.01) (VEGF: vascular endothelial growth factor; GDNF: glial cell line-derived neurotrophic factor).

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  Fig. 5 Proliferation rate of newborn cells in cortex area of ischemic side of rats in each group and at different time points, the method of immunofluorescence staining was used. (A, B) The proliferation of newborn cells was detected in the SVZ area of the ischemic side of rats. Compared with the model group and the sham group, the cell proliferation rate of the BMSCs group was significantly higher than that of the model group and the control group in SVZ area. (C, D) The proliferation of newborn cells was detected in the cortex area of the ischemic side of rats. Compared with the model group and the sham group, the cell proliferation rate of the BMSCs group was significantly higher than that of the model group and the control group in cortex area. (E, F) Proliferation rate of GFAP in SVZ area of ischemic side of rats in each group and at different time points. Compared with the model group and the sham group, the increment rate of astrocytes of the BMSCs group was significantly higher than that of the model group and the control group in SVZ area. (G, H) Proliferation rate of GFAP in cortex area of ischemic side of rats in each group and at different time points. Compared with the model group and the sham group, the increment rate of astrocytes of the BMSCs group was significantly higher than that of the model group and the control group in cortex area (*p<0.05, **p<0.01) (SVZ: Subventricular Zone).

Reference

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