Studies Of Cerebral Protection Of Huo Xue Kai Qiao Granule And Its Clinical Efficacy

Objective: 1.To investigate the clinical efficacy and safety of Huo Xue Kai Qiao-Granule on patients with acute cerebral infarction,providing clinical research evidence for Huo Xue Kai Qiao-Granule’s application.2.To build the middle cerebral artery occlusion(MCAO)on rats,and investigate the infulance of different dose of Huo Xue Kai Qiao-Granule on infarct volume,the degree of improvement of neurological function and brain histopathological changes in MCAO model rats.And then establish the optimal dose of Huo Xue Kai Qiao-Granule,and laid the foundation for the subsequent experiments.3.Discussion Huo Xue Kai Qiao-Granule neuroprotective mechanism of its clinical efficacy may exist,and research the influence of Huo Xue Kai Qiao-Granule on adenosine monophosphate-activated protein kinase(AMPK),brain derived neurophic factor(BDNF)and astrocytes in cerebral infarction area of MCAO model rats at different time points(2h,24 h,and 48h).Materials and Methods: 1.In Tianjin Nankai Hospital Integrative encephalopathy wards,patients with acute cerebral infarction and metting the criteria for admittance row were randomly divided into treatment group and control group,each group contains 40 patients.Patients in treatment group received Huo Xue Kai Qiao-Granule and conventional drug treatment,patients in control group only received conventional drug treatment with the course of 14 days.Before and after treatment,patients were examed through many clinical indicators including neurological deficits,daily living skills,infarct volume,cerebral blood flow and blood rheology.Differences between the two groups before and after treatment of each index were compared.2.Using the classic Zea-Longa suture to establish MCAO model rats.The rats were randomly divided into six groups: Huo Xue Kai Qiao-Granule high-dose group,Huo Xue Kai Qiao-Granule dose group,Huo Xue Kai Qiao-Granule low dose group,positive drug nimodipine group,MCAO model group and the sham-operated group(Sham).The infarct volume and MCAO modeling of stability was measured by 2,3,5-Triphenyl Tetrazolium Chloride(TTC)staining.Then each group pathological brain changes were observed through hematoxylin-eosin staining(HE).The rats were also observed at different time(6h,24 h,48h)by neuromuscular function score.3.Using the classic Zea-Longa suture to build MCAO model rats.SD rats were randomly divided into four groups: Huo Xue Kai Qiao-Granule group,positive drug nimodipine group,MCAO model group and the sham group.Treatment group and positive control group of rats fed before the 3 days modeling were pretreated with drugs.After modeling,the expression of AMPK and p AMPK in the ischemic brain tissue were analysesd using Western Blot and RT-PCR at three different time points(2h,24 h,48h).BDNF,GFAP,and connexin(Cx)in schemic area of MCAO models were measured by immunohistochemical staining.All of these methods were employed to discussion Huo Xue Kai Qiao-Granule neuroprotective mechanism of its clinical efficacy may exist.Results: 1.Compared with the control group,NIHSS and BI score difference was statistically significant in treatment group(P<0.05).After treatment,all the patients had some degree of infarct volume shrinks,in the subgroup of small volume of cerebral infarction(CI),the difference was statistically significant infarct volume(P <0.05)between treatment and control group,but in the large volume CI volume CI subgroup,there was no significant difference between the two groups of patients before and after treatment.2.The MCAO model was successfully built with high stability.Compared with the model group,after 24 h and 48 h of the MCAO model building,significant reduction of the proportion of neurological function and cerebral infarction was observed in Huo Xue Kai Qiao-Granule group(P<0.01).Compared with the model group,HE staining showed that necrotic nerve cells decreased,pyramidal cells become normal morphology,edema lighter nuclei than the rule,especially in treatment group with 35.2g crude drug / kg(high dose group).3.Compared with the MCAO control group,AMPK and p AMPK were low expressed in ischemic region of rat brain tissue of Huo Xue Kai Qiao-Granule group and inmodipine group.AMPK and p AMPK expression levels showed increased gradually in MCAO group from 2h to 48 h.AMPK peaked at 24 h in Huo Xue Kai Qiao-Granule group,and then gradually reduce the amount of expression.p AMPK particles in Huo Xue Kai Qiao-Granule group showed a decreasing trend.Compared with the model group,BDNF content increased significantly in hippocampus from ischemic area in Huo Xue Kai Qiao-Granule group,and over time,the content was increased gradually.In addition,Huo Xue Kai Qiao-Granule can inhibit the excessive proliferation of AS.Compared with the control group at each time point,AS in Huo Xue Kai Qiao-Granule group was significantly decreased.And with the passage of time,astrocytes content showed a decreasing trend in Huo Xue Kai Qiao-Granule group.Huo Xue Kai Qiao-Granule can also inhibit the expression of x43.Compared with the control group,the expression of Cx43 was reduced in treatment group,and with the passage of time,Cx43 showed a increasing trend in Huo Xue Kai Qiao-Granule group.