Optimizing Of Middle Cerebral Artery Occlusion Model In Mice And Investigation Of The Mechanism Of Danhong Intervention

Objective: The purpose of this study is to establish a robust method of focal cerebral ischemia model in different varieties of mice. Then based on the model, the pharmacodynamic evaluation of Danhong injection for the treatment of cerebral ischemia was explored, as well as its mechanism against cerebral ischemia.Methods: Each group of healthy adult KM and C57BL/6 mice were randomly divided into control group(n=10)and Middle cerebral artery occlusion(MCAO) group(n=10).The MCAO model of mice was obtained by intraluminal occlusion using monofilament. Middle cerebral artery blood flow was monitored during the operation.Twenty-four hours after operation,the neurologic function was evaluated,and the infarction volume was calculated by TTC staining to evaluate the reliability of the model.The efficacy evaluation of Danhong injection was performed on 64 male C57 BL / 6 mice, which were divided into the control and the treatment survival group. Six hours after constructing MCAO model,the neurologic function was evaluated,and the infarction volume was calculated. The condition of the treatment survival group for 7 days was observed, and mortality rates was processed.In this study, by using cerebral ischemia mice as models, we adopt new high – through hput strategies of specific enrichment of transcription factors, combining with network pharmacology analyze technology,for comprehensive analysis of relate d transcription factors and signal pathway during the occurrence and development of cerebral ischemia.Results: In the MCAO group, the base value of the cerebral blood flow down of KM and C57 BL / 6 mice respectively was(81.65±4.59)%,(83.68±6.25) %;the neurological deficit score respectively was(2.30±0.82),(2.50±0.80).TTC staining can clearly show the infarction area, and relatively stable, 24 hours of the survival rate of KM and C57 BL / 6 mice were 100% and 80% respectively. The blood pressure has no obvious changes after injection. Compared with control group, the cerebral infarction volume percent dropped 52.6%, 58.87% in different groups,the neurological deficit scores dropped 26.8%, 46.8% respectively.In this study,we find eight key transcription factors including PBX1, ATF1, NFYC, POU3F1, POU3F2, POU3F3, SATB1, SATB2. PBX1,NFYC and ATF1 have direct interaction relations with Danhong forecast targets. HTR2 C and ATP1A1 which interacts with ATF1 is not only forecast targets, but also the known therapeutic targets of stroke. Among the molecules of direct interaction with eight key transcription factors, MAPK1, MAPK11 and JUN are involved in the MAPK and Neurotrophin signaling pathways,while SMAD3, MAPK11 and NR3C1 are involved in NF-kB activated signaling pathways.Conclusion: The modified intraluminal occlusion of MCAO is a kind of reliable and simple method to establish experimental cerebral ischemia model in mice. Danhong injection has a very good efficacy on mice brain ischemia,its mechanism may be related to the regulation the expression of NFYC, POU3 F and STAB.