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
OBJECTIVE: To identify the intervention mechanism of the effect of electroacupuncture on the expression of Ang/Tie-2 m RNA and protein in rats with acute cerebral infarction induced by middle cerebral artery occlusion...OBJECTIVE: To identify the intervention mechanism of the effect of electroacupuncture on the expression of Ang/Tie-2 m RNA and protein in rats with acute cerebral infarction induced by middle cerebral artery occlusion(MCAO).METHODS: Altogether 120 Wistar rats were subjected to MCAO by inserting a nylon filament, andthen divided into 3 groups: control group, injured group and electroacupuncture group. The injured and electroacupuncture groups were further divided into the following 7 subgroups according to the time after MCAO: 3, 6, 12, 24 h, 3, 7 and 12 d, with 8 rats in each subgroup. The electroacupuncture group was given electroacupuncture treatment at Shuigou(GV 26) instantly after operation. The rats were killed at different time points according to their groups, and then the expression levels of Ang/Tie-2 m RNA and protein were detected using Real-Time polymerase chain reaction and immunohistochemical staining.RESULTS: The m RNA and protein expression levels of Ang/Tie-2 in the electroacupuncture group were significant higher than that in the injured group.CONCLUSION: The results suggested that electroacupuncture could significantly regulate the expression of Ang/Tie-2 m RNA and protein in the rats with acute cerebral infarction induced by MCAO, and enhance angiogenesis after ischemic penumbra.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.
基金Supported by Tianjin Natural Science Foundation of Key Projects:Acupuncture Adjustment Th17/Treg Balance Against Nerve Inflammation Damage(No.16JCZDJC36200)And the National Natural Science Foundation of China:Acupuncture Regulation VD Brain Nerve Inflammation and Peripheral Immune Suppression of the Effect and Mechanism(No.81473766)+1 种基金Based on ACE/ACE2 axis and VEGF-Dll4/Notch Pathway to Study the Molecular Mechanism of Acupuncture Intervention in the Establishment of Collateral Circulation of Cerebral Infarction(No.81674056)Molecular Mechanism of Neurotoxicity Induced by Acupuncture in Rats with Cerebral Infarction based on Shh and Wnt Signaling Pathway(No.81473765)
文摘OBJECTIVE: To identify the intervention mechanism of the effect of electroacupuncture on the expression of Ang/Tie-2 m RNA and protein in rats with acute cerebral infarction induced by middle cerebral artery occlusion(MCAO).METHODS: Altogether 120 Wistar rats were subjected to MCAO by inserting a nylon filament, andthen divided into 3 groups: control group, injured group and electroacupuncture group. The injured and electroacupuncture groups were further divided into the following 7 subgroups according to the time after MCAO: 3, 6, 12, 24 h, 3, 7 and 12 d, with 8 rats in each subgroup. The electroacupuncture group was given electroacupuncture treatment at Shuigou(GV 26) instantly after operation. The rats were killed at different time points according to their groups, and then the expression levels of Ang/Tie-2 m RNA and protein were detected using Real-Time polymerase chain reaction and immunohistochemical staining.RESULTS: The m RNA and protein expression levels of Ang/Tie-2 in the electroacupuncture group were significant higher than that in the injured group.CONCLUSION: The results suggested that electroacupuncture could significantly regulate the expression of Ang/Tie-2 m RNA and protein in the rats with acute cerebral infarction induced by MCAO, and enhance angiogenesis after ischemic penumbra.
