The Influence Of Salvia Miltiorrhiza Bunge F.alba On Neurogenesis After Cerebral Ischemia Reperfusion

ObjectiveTo investigate the influence of Salvia miltiorrhiza Bunge f.alba on neurogenesis in ratsafter cerebral ischemia/reperfusion(I/R), and explore the possible mechanism and functionof Salvia miltiorrhiza Bunge f.alba in cerebral ischemic injury in order to provide theexperimental evidence for patients with ischemic cerebrovascular diseases.Methods1. Animal models preparation: The middle cerebral artery occlusion/reperfusion(MCAO) model of rats were established by the modified Longa occlusion method. Maleadult Sprague-Dawley (SD) rats were anesthetized with10﹪chloral hydrate byintraperitoneal injection. The right common carotid artery (CCA), internal carotid artery(ICA) and external carotid artery (ECA) were isolated via a cervical midline incision. Anylon suture was introduced into ECA and advanced into ICA,18.0±0.5mm in depth fromthe CCA branch, via middle cerebral artery, all the ways to the root of anterior cerebralartery (ACA), in order to block the blood of middle cerebral artery. After1hours ofischemia, the nylon suture was drawn out to restore reperfusion for14days. In shamoperation group, the nylon suture was inserted around ECA approaching the ICA branchwithout occlusion.2. BrdU injection: To begin to inject BrdU (100mg/kg) into the enterocoelia threedays before drawing materials, three times a day, with the last injection2hours earlier.3. Salvia miltiorrhiza Bunge f.alba solution preparation: The root was washed anddried, immersed for2hours, boiled for30minutes, then filtered. Three to five times waterwas added then, boiling for20minutes, filtered. The filtrate was then mixed with95%ethanol, precipitating one night for the filtrate. All the above was repeated for three times.The final filtrate was then condensed and reserved at4celsius degrees. 4. Immunohistochemistry to test BrdU positive cells in the brain: Male SD rats wererandomly divided into sham group, I/R group and I/R+Salvia miltiorrhiza Bunge f.albagroup. After ischemia1hour and reperfusion for14days, the rats were killed to preparefrozen sections. Then immunohistochemistry was used to detect BrdU positive cells.5. Water maze test: Five days before the operation, rats were trained for water maze,recording respectively the escape latency before and after ischemia1h/reperfusion14d.Result1. Expression of BrdU positive cells:（1）Neurogenesis in corpora striata of the injured side: The I/R group expressed morethan sham group (p<0.01), but less than I/R+Salvia miltiorrhiza Bunge f.alba group(p<0.01).（2）Neurogenesis in the lateral cerebral ventricle of the injured side: The I/R groupexpressed more than sham group (p<0.01), but less than I/R+Salvia miltiorrhiza Bungef.alba group (p<0.01).（3）Neurogenesis in hippocampus of the injured side: The I/R group expressed morethan sham group (p<0.01), but less than I/R+Salvia miltiorrhiza Bunge f.alba group(p<0.01).2. Water maze test: Escape latency(EL) of I/R group was longer than that of shamgroup (p<0.01), while that of Salvia miltiorrhiza Bunge f.alba was much shorter thanI/R(p<0.01).Conclusion1. Neurogenesis were increased after the acute focal cerebral ischemia injury in rats，brain.2. Based on the first conclusion, neurogenesis was considerably increased with the useof Salvia miltiorrhiza Bunge f.alba, especially in corpora striata, lateral cerebral ventricleand hippocampus.3. The use of Salvia miltiorrhiza Bunge f.alba strengthened the learning and memoryability of rats.