The Effects Of Breviscapine Injection On Cerebral Ischemia Reperfusion Injury Of Rats And The Mechanism Research Induced By Micro Rna And CYP

Objective: To study the mechanism of Breviscapine injections on cerebral ischemia reperfusion injury by micro RNAs,CYP450 and oxidative stress inducing.Medthods:（1）The micro RNAs effects of curatively giving three different doses of Breviscapine injection on cerebral ischemia reperfusion（I/R）injury of rats: 54 male SD rats with SPF level,weight 240³00g,were randomly divided into five groups,as 7 in shame,10 in model,10 in Bre 12mg·kg-1,10 in Bre 6mg·kg-1,10 in Bre 3mg·kg-1.The cerebral ischemia rats were modeled by middle cerebral artery occlusion（MCAO）.The score of ethology,mortality,the expression of micro RNA-27a-3p,-9-5p,-153-5p and Bcl-2 in serum and the expression of micro RNA-27a-3p,-9-5p,-153-5p,-181a-5p,-93-5p,-335,-20a-5p,-290 and Hif-1α and the content of TNF-α,IL^-10 in brain tissue were examined to observe the effect of these three different doses of Breviscapine injections by intravenous injection after 2 hours ischemia and reperfusion immediately,once a day for 7 days.（2）The CYP450 effects of curatively giving three different doses of Breviscapine injection on cerebral ischemia reperfusion（I/R）injury of rats:the experiment subjects,groups and the doses of Breviscapine injection were as same as（1）.The expression of CYP4A1,4A3,4A8,4F4 and the content of e NOS in brain homogenate were examined to observe the effect of these three different doses of Breviscapine injections by intravenous injection after 2 hours ischemia and reperfusion immediately,once a day for 7 days.（3）The anti-oxidative stress effects of curatively giving three different doses of Breviscapine injection on cerebral ischemia reperfusion（I/R）injury of rats:146 male SD rats with SPF level,weight 240³00g,were randomly divided into five groups,as 20 in shame,40 in model,26 in Bre 12mg·kg-1,30 in Bre6mg·kg-1,30 in Bre 3mg·kg-1.The cerebral ischemia rats were modeled by middle cerebral artery occlusion（MCAO）.The score of ethology,the volume of cerebral infarction,mortality,superoxide dismutase（SOD）,malondialdehyde（MDA）,xanthine oxidase（XOD）,nitric oxide synthase（NOS）,constitutive nitric oxide synthase（TNOS）,inducible nitric oxide synthase（i NOS）,catalase cas（CAT）,glutathione（GSH）,glutathione peroxidase（GSH-PX）,glutathione reductase（GR）in serum and brain homogenate were examined to observe the effect by intravenous Breviscapine injection after 2 hours ischemia,and reperfusion immediately,once a day for7 days.Results:（1）Bre12 and Bre6 can effectively reduce behavioral score at 8,24,48,72,96,120,144 and 168 h and the volume of cerebral infarction at 168 h.Bre12 could up-regulate the expression of rno-mi R-290（P<0.05）,-335（P<0.05）,the content of IL^-10（P<0.01）in brain homogenate and reduce the content of TNF-α（P<0.01）.Bre6 had no effects on micro RNA in brain homogenate and serum,but could up-regulate the expression of Bcl-2（P<0.01）,increase the content of IL^-10（P<0.01）and reduce TNF-α（P<0.01）content in brain homogenate.Bre3 could down-regulate the expression of rno-mi R-153-5p（P<0.05）,reduce the content of TNF-α（P<0.01）in brain homogenate and up-regulate the expression of Bcl-2（P<0.01）in serum,but it had on effects on regulating the expression of rno-mi R-9-5p,-27a-3p,-181a-5p,-20a-3p,-290,-335,-93-5p,Bcl-2 and Hif-1α in brain homogenate and serum.（2）Bre12 can down-regular the expression of CYP4A8（P<0.05）in brain homogenate;Bre6 and Bre3 can effectively down-regular the expression of CYP4A3（P<0.01）,4A8（P<0.01）,but Bre3 had no effects on the expression of CYP4A1 and CYP4F4.（3）Bre12 and Bre6 can effectively reduce behavioral score at 8,24,48,72,96,120,144 and 168 h,the volume of cerebral infarction at 168 h and mortality.Bre12 could reduce NOS（P<0.01）in brain homogenate and the content of TNOS（P<0.05）（P<0.01）,i NOS（P<0.01）（P<0.05）,MDA（P<0.01）,XOD（P<0.01）（P<0.05）in brain homogenate and serum.Bre12 could increase the activity of GR（P<0.05）,GSH-PX（P<0.05）,CAT（P<0.05）in brain homogenate and GSH（P<0.05）（P<0.01）content and SOD（P<0.01）（P<0.05）activity in brain homogenate and serum.Bre6 could reduce i NOS（P<0.01）content in serum and SOD（P<0.01）activity in in brain homogenate.Bre6 could increase the content of MDA（P<0.01）（P<0.05）,XOD（P<0.01）,GSH（P<0.05）（P<0.01）in brain homogenate and serum.Bre3 had no effects on NOS,TNOS,i NOS,MDA,GSH content and SOD,GSH-PX,GR,CAT activity in serum and brain homogenate but to reduce content of MDA（P<0.01）and XOD（P<0.01）in brain homogenate and behavioral score at 120h（P<0.05）and 168h（P<0.05）.Conclusion:（1）The Breviscapine injection could reduce the cerebral ischemia reperfusion injury by regulating the expression of micro RNA to impact the inflammation factors and oxidant enzyme.（2）The Breviscapine injection could reduce the cerebral ischemia reperfusion injury by down-regularing the expression of CYP4A3 and 4A8 to reduce the contents of e NOS.（3）Curatively injecting Bre12 and Bre6 Breviscapine could effectively reduce cerebral ischemia reperfusion injury by anti-oxidative stress which has closed relationship with the mechanism of micro RNA and CYP inducing.