Danshen Injection On Rat Cerebral SVZ Stem Cell Proliferation

Objective:To investigate the effects of Danshen injection on promote the proliferation of neural stem cells in the bilateral subventricular zone (SVZ) of rats after mobilization with recombinant human granulocyte colony stimulating factor (rhG-CSF) after cerebral infarction, and to explore it’s mechanism, illustrate the possible pathway for the Danshen injection combined with rhG-CSF to treat the cerebral infarction.Methods:The48successful middle cerebral artery occlusion (MCAO) adult male Sprague-Dawley (SD) rat models were randomly devided into a rhG-CSF mobilization+Danshen injection group, a Danshen injection group, a rhG-CSF mobilization group, a saline group. In each group,12rats were randomly allocated into groups. Each group according to experimental purpose is divided into TTC staining group and frozen section group.(6rats in each subgroups); and randomly selected12normal rats served as sham operation group. Neurological deficits were evaluated using the modified neurological severity score (mNSS) before treatment and on the7th day after treatment. TTC staining group rats were used to detected infarction area; BrdU/nestin, GFAP/nestin cells in the SVZ were detected by immunofluorescence techniques in frozen section group, and were compared between groups. BrdU/NeuN positive cells were detected in rhG-CSF mobilization+Danshen injection group and Danshen injection group. Result:1. Except for the sham operation group, each group of rats’s mNSS scores were lower than before used drug administration, the difference was statistically significant (P<0.05); RhG CSF mobilization+Danshen injection group rats’s mNSS score was decreased compared with Danshen injection group, rhG-CSF mobilization group and saline group, the difference was statistically significant (P<0.05); Danshen injection group has no significant difference compared with rhG-CSF mobilization group(P>0.05); the two groups compared with the saline group, the difference was statistically significant (P<0.05).2. TTC staining group rats showed no focal infarction in sham operation group; after used the drugs according to the experimental purpose, we found that the infarction area in rhG-CSF mobilization+Danshen injection group was significantly decreased compared with the other three groups.3. In each group, the number of Brdu+cells was no statistical difference in bilateral SVZ (P>0.05); the number of BrdU+/nestin+cells in sham operation group was lower than the other of groups in the SVZ, the difference was statistically significant (P<0.05); The number of BrdU+/nestin+cells in rhG-CSF mobilization+Danshen injection group was increased significantly than the other groups,, the difference was statistically significant (P<0.05); Danshen injection group compared with rhG-CSF mobilization group has no difference (P>0.05), the two group’s BrdU+/nestin+cells were increased compared with the saline group, and the difference was statistically significant (P>0.05); in each group, the ratio of BrdU+/nestin+cells count of BrdU1+cells had no statistical difference in bilateral SVZ (P>0.05); there han’t found GFAP+/nestin+ cells in the bilateral SVZ.4. In rhG-CSF mobilization+Danshen injection group and Danshen injection group, a large number of BrdU+/NeuN+cells were detected in the bilateral SVZ.Conclusion:1. With the passage of time, the neural function were improved after cerebral infarction in rats; used rhG-CSF alone or Danshen injection can improve the neural function improvement; rhG-CSF combined with Danshen injection can significantly improve the neural function in rats with cerebral infarction.2. Rats after cerebral infarction, the number of NSCs proliferation in the SVZ.3. Used rhG-CSF alone or Danshen injection can promote the number of NSCs proliferation in the SVZ; rhG-CSF combined with Danshen injection can significantly promote the number of NSCs proliferation, and make the more NSCs differentiation into neurons.