Effects Of Buyang Huanwu Decotion Combined With BMSCs Transplantation On Notch Signaling Pathway In Rats With Cerebral Ischemia

Objective: By setting up a cerebral ischeima rat model of middle cerebral artery occlusion,to observe the scores of neurological deficits,pathological changes,and Notch signaling pathways in rats with cerebral ischemia after Buyang Huanwu Decoction combined with BMSCs transplantation,to study the changes of Notch signaling pathway after cerebral ischemia,and to explore whether the neuroprotective effect of Buyang Huanwu Decoction and BMSCs transplantation on cerebral ischemia is related to Notch signaling pathway,which may provide a new approach for the treatment of cerebral ischeima.methods:The animals were randomly divided into 5 groups,the rat model of middle cerebral artery occlusion was established by suture emboli method,divided into blank group,model group,Buyang Huanwu Decoction group,BMSCs transplantation group,Buyang Huanwu Decoction and BMSCs transplantation combined group.BMSCs were extracted by Whole bone marrow culture method,and then cultured and passaged until the cell morphology basically becomes slender spindle shape,Identified by flow cytometry,labeled with Brdu before trasplantation.BMSCs were transplanted through the tail vein.Brdu-DCX immunofluorescence double-label detection was performed on the transplantation group and the combination group.The neurological deficit score was evaluated by Zea-Longa method.The pathological changes of the ischemic area were observed by HE staining,immunohistochemistry method was used to determine the positive expression of the Notch signaling pathway Notch1 ? Hes1 and Jagged1 in the ischemic parts of the brain tissue.Results:(1)Results of neurological deficit score: At the 24 h time point,compared with the model group,the scores of the combined group decreased,and the difference was statistically significant(P<0.05)..At 7d and 14 d time point,compared with the model group,the Buyang Huanwu Decoction group,BMSCs transplantation group,the scores of the combined group decreased significantly(P<0.05);at the 14 d time point,compared with the BMSCs transplantation group,the scores in the Buyang Huanwu Decoction group decreased significantly(P<0.05).(2)HE staining showed that,At 24 hours,The arrangement of cells was loose,the number was decreased,the morphology was abnormal,there were vacuoles and triangles,and the nuclear condensation,interstitial staining was sparse,and the cytoplasm stained deeper.Compared with the model group,the Buyang Huanwu Decoction group,the BMSCs transplantation group and the combined group all reduced the abnormal morphological changes to varying degrees.By the 7th and 14 th day,the pathological abnormalities of each group gradually decreased,but the combined The group's cell morphology improved more significantly.(3)Notch signaling pathway immunohistochemistry results:(1)The positive cell expression of notch1 in each group:At 24 h,7d,and 14 d,compared with the blank group,the positive expression of Notch1 protein in the ischemic region of every model rats increased significantly(p<0.05);compared with the model group,the number of positive cells of the combined group increased significantly(p<0.05);Compared with Buyang Huanwu Decoction group and BMSCs transplantation group,the combined group increased significantly(p<0.05);at 7d and 14 d time points,compared with the model group,the number of positive cells in the Buyang Huanwu Decoction group and the BMSCs transplantation group increased significantly(p<0.05).(2)The positive cell expression of jagged1 in each group:At 24 h,7d,and 14 d,compared with the blank group,the positive expression of jagged1 protein in the ischemic area of every model rats increased significantly(p<0.05);Compared with the the model group,Buyang Huanwu Decoction group and the BMSCs transplantation group,the number of positive cells in the combined group increased significantly(p<0.05);at the 7d and14 d,compared with the BMSCs transplantation group,the number of positive cells in the Buyang Huanwu Decoction group increased significantly(p<0.05);compared with the model group,the number of positive cells in the Buyang Huanwu Decoction group and the BMSCs transplantation group increased significantly(p<0.05).(3)The positive cell expression of jagged1 in each group:At 24 h,7d,and 14 d,compared with the blank group,the positive expression of jagged1 protein in the ischemic area of every model rats increased significantly(p<0.05);Compared with the the model group,Buyang Huanwu Decoction group and the BMSCs transplantation group,the number of positive cells in the combined group increased significantly(p<0.05);Compared with the the BMSCs transplantation group,the number of positive cells in the Buyang Huanwu Decoction group group increased significantly(p<0.05);at the 7d and14 dcompared with the model group,the number of positive cells in the Buyang Huanwu Decoction group and the BMSCs transplantation group increased significantly(p<0.05).conclusions: Buyang Huanwu Decoction combined with BMSCs transplantation in treatment of cerebral ischemia can promote the recovery of pathological changes and neurological deficits in cerebral ischemia.The mechanism may be related to the increase in the expression level of Notch1,hes1 and jagged1 and promote the proliferation of neural stem cells.