Preconditioning of bone marrow-derived mesenchymal stromal cells by tetramethylpyrazine enhances cell migration and improves functional recovery after focal cerebral ischemia in rats

Abstract

BackgroundTransplantation of bone marrow-derived mesenchymal stem cells (BMSCs) is one of the new therapeutic strategies for treating ischemic stroke. However, the relatively poor migratory capacity of BMSCs toward infarcted regions limited the therapeutic potential of this approach. Pharmacological preconditioning can increase the expression of CXC chemokine receptor 4 (CXCR4) in BMSCs and enhance cell migration toward the injury site. In the present study, we investigated whether tetramethylpyrazine (TMP) preconditioning could enhance BMSCs migration to the ischemic brain and improve functional recovery through upregulating CXCR4 expression.MethodsBMSCs were identified by flow cytometry analysis. BMSCs migration was evaluated in vitro by transwell migration assay, and CXCR4 expression was measured by quantitative reverse transcription-polymerase chain reaction and western blot analysis. In rats with focal cerebral ischemia, the neurological function was evaluated by the modified neurological severity score, the adhesive removal test and the corner test. The homing BMSCs and angiogenesis were detected by immunofluorescence, and expression of stromal cell-derived factor-1 (SDF-1) and CXCR4 was measured by western blot analysis.ResultsFlow cytometry analysis demonstrated that BMSCs expressed CD29 and CD90, but not CD34 and CD45. TMP pretreatment dose-dependently induced BMSCs migration and CXCR4 expression in vitro, which was significantly inhibited by AMD3100, a CXCR4 antagonist. In rat stroke models, we found more TMP-preconditioned BMSCs homing toward the infarcted regions than nonpreconditioned cells, leading to improved neurological performance and enhanced angiogenesis. Moreover, TMP-preconditioned BMSCs significantly upregulated the protein expression of SDF-1 and CXCR4 in the ischemic boundary regions. These beneficial effects of TMP preconditioning were blocked by AMD3100.ConclusionTMP preconditioning enhances the migration and homing ability of BMSCs, increases CXCR4 expression, promotes angiogenesis, and improves neurological performance. Therefore, TMP preconditioning may be an effective strategy to improve the therapeutic potency of BMSCs for ischemic stroke due to enhanced BMSCs migration to ischemic regions.

Tetramethylpyrazine Protects Bone Marrow-Derived Mesenchymal Stem Cells against Hydrogen Peroxide-Induced Apoptosis through PI3K/Akt and ERK1/2 Pathways

“…Passage 3 (P3) BMSCs showed typical spindle-shaped cell morphology. 13) P3 BMSCs were used for the following experiments. The cells were pretreated with TMP (10, 25, 50, 100, and 200 µmol/L) for 24 h and then treated with H 2 O 2 (500 µmol/L) for 24 h. 14,15) Wortmannin (an inhibitor of PI3K, 200 nmol/L) and PD98059 (an inhibitor of ERK1/2, 25 µmol/L) were incubated for 1 h after TMP pretreatment.…”

“…13) Briefly, the bone marrow was flushed out with no serum DMEM/F12. The cell suspension was plated in DMEM/F12 medium plus 10% FBS, 1% penicillin/streptomycin at 37°C in a humidified atmosphere containing 5% CO 2 and 95% O 2 .…”

Nanomorphological and mechanical reconstruction of mesenchymal stem cells during early apoptosis detected by atomic force microscopy

“…Newly generated cells derived from BMSCs could replace dead cells in injured hearts and vessels. However, BMSC transplantation and engineered tissue construction are hampered by poor survival of stem cells in the recipient sites [Wang et al, 2013;Li et al, 2017]. Apoptosis has become a bottleneck for the further development of stem cell transplantation and tissue engineering.…”

Preconditioning with far-infrared irradiation enhances proliferation, cell survival, and migration of rat bone marrow-derived stem cells via CXCR4-ERK pathways

“…The present study also does not provide evidence as to whether FIR-preconditioned BMSCs can preferentially migrate to damaged cardiac tissue. Several reports have suggested that CXCR4-enhanced BMSC preconditioning may beneficially effect the mobilization or transplantation of cells into damaged areas in rats 24 – 26 . For example, preconditioning with tetramethylpyrazine (TMP) significantly upregulated the protein levels of SDF-1 and CXCR4 in BMSCs 24 .…”