Objective To observe the angiogenesis effect of electroacupuncture(EA)at Shuigou acupoint(GV 26)in the treatment of cerebral ischemia,and explore the value of miRNA-7(miR-7)in it.Methods First,48 mice were randomly di...Objective To observe the angiogenesis effect of electroacupuncture(EA)at Shuigou acupoint(GV 26)in the treatment of cerebral ischemia,and explore the value of miRNA-7(miR-7)in it.Methods First,48 mice were randomly divided into sham operation,middle cerebral artery occlusion(MCAO)model,and EA treatment groups.Then 9 mice were divided into carrier control group,miR-7 knockout group and miR-7 overexpression group(n=3 each group).Finally,20 mice were divided into model and carrier control group,model and miR-7 knockout group,EA treatment and carrier control group and EA treatment and miR-7 overexpression group,with 3–6 mice in each group.The MCAO model was established in the MCAO and EA groups.Neurological deficit score and 2,3,5-triphenyltetrazolium chloride(TTC)staining were used to evaluate the severity of cerebral ischemia.Hematoxylin-eosin staining was used to describe basic pathological changes.Immunohistochemistry was used to quantify cerebral microvessel density.Real-time PCR and Western blot were used to detect the expression of miR-7 and its downstream target genes Krüppel-like factor 4/vascular endothelial growth factor(KLF4/VEGF)and angiopoietin-2(ANG-2)in the ischemic cerebral cortex.Results After EA,neurological deficit scores and infarction volumes decreased,and the density of cerebral microvessels increased.In the MCAO group,miR-7 expression was higher than that in the sham group(P<0.01).After EA at GV 26,miR-7 expression decreased(P<0.01)and the expression of downstream target genes KLF4/VEGF and ANG-2 increased as compared with the MCAO group(P<0.01).After EA combined with overexpression of miR-7,the expression of downstream target genes KLF4/VEGF and ANG-2 decreased compared to the control EA group(P<0.01).After miR-7 knockdown,the expression of KLF4/VEGF and ANG-2 increased(P<0.05 or P<0.01).Conclusions EA could promote angiogenesis in MCAO mice likely by inhibiting the expression of miR-7 and relieving inhibition of downstream target genes KLF4/VEGF and ANG-2.展开更多
基金Supported by the National Natural Science Foundation of China(No.81874505)。
文摘Objective To observe the angiogenesis effect of electroacupuncture(EA)at Shuigou acupoint(GV 26)in the treatment of cerebral ischemia,and explore the value of miRNA-7(miR-7)in it.Methods First,48 mice were randomly divided into sham operation,middle cerebral artery occlusion(MCAO)model,and EA treatment groups.Then 9 mice were divided into carrier control group,miR-7 knockout group and miR-7 overexpression group(n=3 each group).Finally,20 mice were divided into model and carrier control group,model and miR-7 knockout group,EA treatment and carrier control group and EA treatment and miR-7 overexpression group,with 3–6 mice in each group.The MCAO model was established in the MCAO and EA groups.Neurological deficit score and 2,3,5-triphenyltetrazolium chloride(TTC)staining were used to evaluate the severity of cerebral ischemia.Hematoxylin-eosin staining was used to describe basic pathological changes.Immunohistochemistry was used to quantify cerebral microvessel density.Real-time PCR and Western blot were used to detect the expression of miR-7 and its downstream target genes Krüppel-like factor 4/vascular endothelial growth factor(KLF4/VEGF)and angiopoietin-2(ANG-2)in the ischemic cerebral cortex.Results After EA,neurological deficit scores and infarction volumes decreased,and the density of cerebral microvessels increased.In the MCAO group,miR-7 expression was higher than that in the sham group(P<0.01).After EA at GV 26,miR-7 expression decreased(P<0.01)and the expression of downstream target genes KLF4/VEGF and ANG-2 increased as compared with the MCAO group(P<0.01).After EA combined with overexpression of miR-7,the expression of downstream target genes KLF4/VEGF and ANG-2 decreased compared to the control EA group(P<0.01).After miR-7 knockdown,the expression of KLF4/VEGF and ANG-2 increased(P<0.05 or P<0.01).Conclusions EA could promote angiogenesis in MCAO mice likely by inhibiting the expression of miR-7 and relieving inhibition of downstream target genes KLF4/VEGF and ANG-2.
