Tetramethylpyrazine Promotes Migration of Neural P

Authors: Xiangying Kong Micun Zhong Xiaohui Su Qingxia Qin Hongchang Su Hongye Wan Cuiling Liu Jiajia Wu Hongcai Shang Yanjun Zhang Na Lin

Publish Date: 2015/11/28

Volume: 53, Issue: 9, Pages: 6526-6539

PDF Link

Abstract

Stem cell therapy may provide a novel therapeutic method for the replacement and regeneration of damaged neural cells in the central nervous system However insufficient stem cell migrating into the injured regions limits its applications Although tetramethylpyrazine TMP originally isolated from Ligusticum walliichi Chuanxiong has been widely used to treat ischemic stroke in the clinic for many years because of its role in neuroprotection how TMP impacts the migration of neural progenitor/precursor cells NPCs and what is the underlying cellular and molecular mechanism remain largely unknown Here we found that TMP promoted NPC migration through increasing the expression and secretion of stromal cellderived factor 1 SDF1 a chemokine that has been well demonstrated to direct NPC cell trafficking in a dosedependent fashion as analyzed by using different methods The role of TMP in NPC migration could be inhibited by AMD 3100 a chemokine CXC motif receptor 4 CXCR4 antagonist Further investigation of the molecular mechanisms revealed that TMP treatment rapidly activated phosphatidylinositol 3kinase PI3K/Akt protein kinase C PKC and extracellular signalregulated kinase ERK but not Pyk2 in NPCs NPC migration could be blocked by using pharmacological inhibitors for these signaling pathways such as LY294002 a PI3K inhibitor MyrψPKC a PKC inhibitor and an ERK1/2 inhibitor Furthermore TMP enhanced NPC migration toward the ischemic region in the MCAO rat model Our findings provide mechanistic insights into the role of TMP in treating the neuropathological diseases which suggest that TMP may be used as a potent drug for improving NPC migration in stem cellbased therapyThe investigation was supported by Natural Science Foundation of China 30873394 the Fundamental Research Funds for the Central Public Welfare Research Institutes ZZ070825 ZXKT15014 National Major Scientific and Technological Special Project for ‘‘Significant New Drugs Creation’’ No 2013ZX09301307Involvement of the SDF1 and PI3K pathway in NPCs migration induced by TMP in vitro a NPCs were pretreated with AMD 3100 5 μg/mL for 2 h then incubated with LY294002 10 μM 32Aminoethyl54ethoxyphenylmethylene 24 thiazolidinedione hydrochloride ERK inhibitor 10 μM MyrψPKC 10 μM and PF431396 25 μM for 30 min at 37 °C followed by exposing to TMP 50 μg/mL for another 24 h Cell migration was monitored under a microscope The images are representative of three independent experiments b The data are expressed as the mean ± SD of three independent experiments P  001P  0001 versus Control group P  005 versus AMD+TMP group GIF 143 kbInvolvement of the SDF1 and PI3K pathway in NPCs migration induced by TMP in vivo a Schematic representation of the time course of experimental design Adults SD rats were subjected to cerebral ischemia operation then treated with or without TMP 40 mg/kg in the presence of AMD 3100 1 mg/kg or LY294002 10 mg/kg for 3 day Neurological deficit score b and infarction volume by TTC staining c in MCAO rats were shown Cells migration was assessed by showing BrdU green and DCX red doublelabeled cells d The microphotographs showed BrdU+/DCX+ cells at day 21 from the corpus striatum CS and the subgranular zone SGZ Quantitation of BrdU+/DCX+ cells in the CS e SGZ f and SVZ g at day 7 and 21 after MCAO Data are represented as the mean ± SD n = 48 P  005 P  001 versus sham group P  005 P  001 versus model group P  005 P  001 versus TMP only group GIF 102 kb

Keywords: