Effects Of Shouwuyizhi Capsule On Angiogenesis In Vascular Cognitive Impairment

Objective:(1) To observe the effects on proliferation,migration, capillary-like structures ofHUVECs and expresstion of VEGF,Ang-1,Tie-2,Notch4treatment with ShouWuYiZhicapsule,then to study the effect of ShouWuYiZhi capsule on angiogenesis and investigatethe mechanisms of ShouWuYiZhi capsule.(2)To investigate the effect ShouWuYiZhi capsule on angiogenesis through observingthe expression of CD34of VCI model rats.(3) To investigate the potential mechanisms of ShouWuYiZhi capsule on angiogenesisthrough observing the expression of the genes of VEGF,Ang-1,Tie-2,Notch4of VCI modelrats.Methods:(1) At the cellular level,HUVECs were cultured,we observed the catalpol effects onHUVECs prolifersion,migration,capillary-like structures using cell counting,MTT,respectively.In addition,the expression of the genes of VEGF,Ang-1,Tie-2,Notch4onHUVECs induced by ShouWuYiZhi capsule were observed with real time PCR.(2)Rats were used and the VCI model was established with MCAO.80rats werescreened after Morris water maze test and randomly divided into4groups: sham operationgroup, model group, Nimodipine group and Shouwuyizhi Capsule group, and each grouphas20rats. After4-week administration, behavioral change of rat was recorded by Morriswater maze. Ultrastructure alternation of vascular endothelial cells was observed viatransmission electric microscope.CD34expression was measured by immunohistochemistrymethod and the expression of the genes of VEGF,Ang-1,Tie-2,Notch4were measured using real time PCR.Results:1. Results in vitro(1) ShouWuYiZhi capsule caused a significant increase in MTT test of HUVECs(OD(0.99±0.03) vs (0.54±0.02) in control group (p <0.01)). The incubation withShouWuYiZhi capsule led to a significantly augmented migratory rate(94.83%vs70.94%in cont rol group (p <0.01)).ShouWuYiZhi capsule induced the differentiation ofendothelial cells into capillary-like structures ((23.42±2.83) mm vs (15.23±3.44)mm incontrol group (p <0.01)).(2) The content of cerebral tissue of angiogenesis factors (VEGF,Ang-1,Tie-2,Notch4)of rats model with vascular congnitive impairment were higher than the rats of sham group(p <0.01);After the treatment,the content of theses factors were further highter thanmodel group(p <0.01, p <0.05).Except VEGF,Tie-2,the content of Ang-1,Notch4ofShouwuyizhi group were highter than Nimodipine group(p <0.05).2. Results in vivo(1)The results of Morris water maze showed that assessment of orientation test andspatial probe test from ShouWuYiZhi capsule group were significantly higher than modelgroup and Nimodipine group (p <0.01, p <0.05).(2) About the microvessel dencity of ischemia cerebral tissue,all of the other threegroups were higher than sham group with the significant difference (p <0.01, p <0.05)；Compared with model group, Nimodipine group and Shouwuyizhi group had thesignificant difference (p <0.01, p <0.05)；Shouwuyizhi group had no difference withNimodipine group(p>0.05).(3) The changes of BCECs microstructure were observed by transmission electronmicroscopy.Compared with sham group and model group, ShouWuYiZhi capsule reducedthe BCECs edema significantly.The number of chondriosome in ShouWuYiZhi capsulegroup is more than that of in model group, and near normal levels.(4)The content of cerebral tissue of angiogenesis factors (VEGF,Ang-1,Tie-2,Notch4)of rats model with vascular congnitive impairment were higher than the rats of sham group(p <0.01); After the treatment,the content of theses factors were further highter thanmodel group(p <0.01, p <0.05).Except Tie-2,the content of VEGF,Ang-1,Notch4of Shouwuyizhi group were highter than Nimodipine group(p <0.05).Conclusion:(1) ShouWuYiZhi capsule enhences endothelial cell prolifersion,stimulatesendothelial cell migration,capillary-like structures accompanied by upregulating theexpression of the genes of VEGF, Ang-1, Tie-2,Notch4.(2) ShouWuYiZhi capsule given after pMCAO in adult rats provides significantneuroprotection and angiogenesis,mediated partly by activation of the the expression ofthe genes of VEGF, Ang-1, Tie-2,Notch4.(3) ShouWuYiZhi capsule could improve the learning and memory ability of VCImodel rats, and ShouWuYiZhi capsule is a valid therapy to VCI,which might be moreeffective than Nimodipine.