基于Notch/血小板源性生长因子B通路探讨藏医金针促进缺血性卒中大鼠血管新生的机制OA

Objective To observe the effect of Tibetan golden acupuncture on angiogenesis and the expression of Notch/PDGF-B pathway proteins in MCAO rats,so as to explore the mechanism of Tibetan golden acupuncture in promoting angiogenesis.Methods SD rats were randomly divided into sham-operation group,a model group,a Tibetan golden acupuncture group,and a Tibetan golden acupuncture plus inhibitor group,with 18 rats in each group.The MCAO model was established by Zea Longa method.The Tibetan golden acupuncture group and the Tibetan golden acupuncture+inhibitor group were treated once a day for seven consecutive times,and the Notch pathway inhibitor DAPT was given in-traperitoneal injection in the Tibetan golden acupuncture+inhibitor group.The sham-operation group and the model group were handled in parallel for the same duration without acupuncture.modified neurological severity scores(mNSS)score was used to detect the degree of neurological deficit,laser speckle contrast imaging was used to detect cerebral blood flow,TTC staining enabled the detection of cerebral infarction volume,immunohistochemistry was used to detect the number of platelet-derived growth factor B(PDGF-B)-positive cells.The protein expressions of platelet-derived growth factor receptor B(PDGFR-B),DLL-4,Jagged-1,Hey1,Hey2,Hes1,and Notch1 were detected by Western blot.PDGFR-B,DLL-4,Jagged-1,Hey1,Hey2,Hes1,Notch1 protein expression,and the mRNA expression of PDGFR-B,Hey1,Hey2,Hes1 were detected by RT-PCR.Results Compared with the sham-operation group,the ratio of left and right cerebral blood flow was decreased(P<0.01),TTC showed obvious cerebral infarction,and the number of PDGF-B-positive cells was increased in the model group(P<0.05).The protein expression of PDGFR-B,DLL-4,Jagged-1,Hey1,Hey2,Hes1 and Notch1 was increased(P<0.05 or P<0.01).The corresponding mRNA expressions of PDGFR-B,Hey1,Hey2 and Hes1 were increased(P<0.01).Compared with the model group,the mNSS score was decreased(P<0.01),the ratio of left and right cerebral blood flow was increased(P<0.01),the cerebral infarction volume was decreased(P<0.01),and the number of PDGF-B positive cells was increased(P<0.01)in the Tibetan golden acupuncture group.The protein expressions of PDGFR-B,DLL-4,Jagged-1,Hey1,Hey2,Hes1 and Notch1 were increased(P<0.01).The corresponding mRNA expressions of PDGFR-B,Hey1,Hey2 and Hes1 were in-creased(P<0.01).Compared with the model group,no significant differences were observed in the ratio of left and right cerebral blood flow,cerebral infarction volume,or the protein and mRNA expressions of PDGFR-B,DLL-4,Jagged-1,Hey1,Hey2,Hes1,and Notch1 in the Tibetan golden acupuncture plus inhibitor group(P>0.05),although the mNSS score was reduced(P<0.01).No significant differences were found in the mRNA expressions of PDGFR-B,Hey1,Hey2,and Hes1(P>0.05).Compared with the Tibetan golden acupuncture group,the mNSS score was increased(P<0.01),the ratio of left and right cerebral blood flow was decreased(P<0.05),the cerebral infarction volume was increased(P<0.01),and the number of PDGF-B positive cells was decreased(P<0.01)in the Tibetan golden acupuncture+inhibitor group.PDGFR-B,DLL-4,Jagged-1,Hey1,Hey2,Hes1,Notch1 protein expression decreased(P<0.01).The corresponding mRNA expressions of PDGFR-B,Hey1,Hey2 and Hes1 were decreased(P<0.01).Conclusion Tibetan golden acupuncture can upregulate the expressions of DLL-4,Jagged-1,Hey1,Hey2,Hes1 and Notch1 proteins in the Notch pathway,increase the expressions of PDGF-B and PDGFR-B,promote angiogenesis,increase cerebral blood flow on the affected side,reduce the infarct volume,and alleviate neurological damage in MCAO rats.