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OBJECTIVE@#To explore the effect of electroacupuncture (EA) intervention on the vasoconstriction of cerebral artery smooth muscle cells after cerebral infarction.@*METHODS@#Male Wistar rats were randomly divided into 3 groups by a random number table: the model group (n=24), the EA group (n=24), and the normal group (n=6). The model and the EA groups were divided into different time subgroups at 0.5, 1, 3, and 6 h after middle cerebral artery occlusion (MCAO), with 6 rats in each subgroup. MCAO model was established using intraluminal suture occlusion method. The EA group was given EA treatment at acupoint Shuigou (GV 26) instantly after MCAO for 20 min. The contents of cerebrovascular smooth muscle MLCK, the 3 subunits of myosin light chain phosphatase (MLCP) MYPT1, PP1c-δ and M20, as well as myosin-ATPase activity were detected using immunohistochemistry and Western blotting.@*RESULTS@#The overall expression level of the MYPT1 and PP1c-δ in the model group was significantly higher (P<0.01). After EA intervention, the 0.5 h group expression level was close to that of the normal group (P>0.05), and the other subgroups were still significantly higher than the normal group (P<0.01). After EA intervention, the expression level of each subgroup was significantly lower than the corresponding model group. There was a significant difference between the 0.5 and 1 h subgroups (P<0.01), while a difference was also observed between the 3 and 6 h subgroups (P<0.05). The dynamic change rule gradually increased with the prolongation of infarction time within 6 h after infarction.@*CONCLUSION@#EA intervention can inhibit contraction of cerebral vascular smooth muscle cells and regulate smooth muscle relaxation by regulating MLCK pathway.
Subject(s)
Rats , Male , Animals , Rats, Wistar , Electroacupuncture , Cerebral Infarction/metabolism , Muscle, Smooth , Acupuncture Points , Brain Ischemia/therapyABSTRACT
OBJECTIVE@#To investigate the effect of electro-acupuncture (EA) on vasomotor symptoms in rats with acute cerebral infarction, by observing the changes in the expression of factors related to the phosphatidylinositol (PI) system.@*METHODS@#Forty-two Wistar rats were randomly divided into 3 groups by a random number table: the control group (n=6), the model group (n=18) and the EA group (n=18). The EA group was given EA treatment at Shuigou (GV 26) instantly after modeling with middle cerebral artery occlusion (MCAO) method, while the model and control groups were not given any treatment. The degrees of neurological deficiency were evaluated using neurological severity scores (NSS) and the brain blood flow was evaluated by a laser scanning confocal microscope. Western blot analysis was conducted to detect the expression levels of G-protein subtype (Gq) and calmodulin (CaM). Competition for protein binding was conducted to detect the expression level of inositol triphosphate (IP3). Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol (DAG). The expression level of intracellular concentration of free calcium ion ([Ca@*RESULTS@#The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO (P<0.01), while the EA group was significantly lower than the model group at 6 h (P<0.01). The cerebral blood flow in the model group was significantly lower than the control group at 1, 3 and 6 h after MCAO (P<0.01), while for the EA group it was remarkably higher than the model group at the same time points (P<0.01). The expressions of Gq, CaM, IP3, DAG and [Ca@*CONCLUSION@#EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction, improve cerebral autonomy movement, and alleviate cerebral vascular spasm.
Subject(s)
Animals , Rats , Acupuncture Therapy , Brain Ischemia , Cerebral Infarction/therapy , Electroacupuncture , Phosphatidylinositols , Rats, WistarABSTRACT
OBJECTIVE: To explore the mechanism of electroacupuncture (EA) underlying improvement of cerebral infarction (CI) by investigating its influence on expression of cerebral Wnt7a, lymphoid enhancer factor-1 (LEF1), glycogen synthase kinase 3β(GSK-3β) and Dickkopf-1(DKK1) mRNA and proteins in CI rats. METHODS: A total of 280 male Wistar rats were randomly divided into blank control (n＝10), sham-operation, model and EA groups，and 90 rats of the last 3 groups were further divided into 1, 3, 6, 9, 12 and 24 h, and 3, 7 and 12 d subgroups with 10 rats in each subgroup. The CI model was established by occlusion of the middle cerebral artery (MCAO). The sham-operation group received the same surgical operation but without thread embolus insertion. EA (2 Hz/15 Hz, 2 mA) was applied to "Shuigou" (GV26) for 20 min, once a day for 1, 3, 7 and 12 d, respectively. The neurological deficit was evaluated by using Neurological Severity Scores (NSS). The expression levels of Wnt7a，LEF1, GSK-3β and DKK1 mRNAs and proteins in the right ischemic brain tissues were detected by Quantative real-time PCR and Western blot, respectively. RESULTS: After MCAO, the NSS score was significantly increased in the model and EA groups relevant to the blank control and sham-operation groups (P0.05). CONCLUSION: EA of GV26 can significantly improve the neurological deficit symptoms in MCAO rats, which may be associated with its effects in up-regulating the expression of Wnt7a and LEF1 mRNAs and proteins, and in down-regulating the expression of GSK-3β and DKK1 mRNAs and proteins.
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OBJECTIVE: To observe the change of neovascular morphology and angiogenesis related factors in the ischemic cerebral area after cerebral infarction and the intervention effect of electroacupuncture (EA). METHODS: Wistar rats were randomly divided into model group(n=90), EA group(n=90), sham operation group(n=90) and control group(n=10). The first three groups were further divided into 1 h, 3 h, 6 h, 9 h, 12 h, 24 h, 3 d, 7 d and 12 d subgroups(n=10 in each subgroup). The cerebral infarction model was established by middle cerebral artery occlusion(MCAO). EA(15 Hz, 2 mA) was applied to "Shuigou"(GV26) for 20 min in the EA group. The 1, 3, 6, 9, 12, 24 h subgroups were treated immediately after modeling, the 3, 7, 12 d subgroups were treated once daily for 3, 7 or 12 days. The neovascular endothelial cells were displayed by immunofluorescence double labeling staining. Quantitive real-time PCR and Western blot were applied to detect the changes of basic fibroblast growth factor (bFGF), angiogenin (Ang) -1, 2, platelet-derived growth factor b (PDGF-b) in ischemic brain tissue, respectively. RESULTS: After modeling, CD31 and Ki67 positive cells were first observed at 24 h in the model group, and reached the peak at 3 d, decreased at 7 d. While in the EA group, the CD31 and Ki67 positive cells were first observed at 12 h, and reached the peak at 3 d, and gradually decreased until 12 d. Compared with the control group, the mRNA expressions of bFGF at 9 h-12 h, Ang-1 at 12 h-12 d, Ang-2 at 1 h-12 d and PDGF-b at 1 h, 6 h, 9 h, 24 h-12 d were increased in the model group(P<0.01, P<0.05). After EA, the mRNA expressions of bFGF at 24 h-12 d, Ang-1 at 3 d-12 d, Ang-2 at 3 h-24 h and PDGF-b at 3 h, 6 h, 3 d-12 d were increased(P<0.05, P<0.01). In comparison with the control group, the proteins of bFGF at 24 h, Ang-1 at 6 h-12 d, Ang-2 at 1 h-12 d and PDGF-b at 1 h-7 d were increased in the model group(P<0.05, P<0.01). After EA, the proteins of bFGF at 3 d-12 d, Ang-1 at 3 d-12 d, Ang-2 at 3 h-12 h and PDGF-b at 6 h, 3 d-12 d were increased compared with the model group(P<0.05, P<0.01). CONCLUSION: EA can up-regulate the expression of angiogenesis-related factors in MCAO rats, which has an important role in the establishment of blood vessel regeneration and collateral circulation, and thus promote the recovery of neurological function.
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Objective To summarize the rules in the application of Shuigou (GV 26) by sorting the ancient literatures involving Shuigou (GV 26).Method A database was established based on systemically rigidly sorting out the indications, paired acupoints, indications of paired acupoints, acupuncture and moxibustion methods, and stimulation dose of Shuigou (GV 26) by retrieving the 1000 books indexed byZhong Hua Yi Dian (Chinese Canon of Medicine) (4th edition).Result A total of 425 items about the indications of Shuigou (GV 26) were concluded, involving 171 ancient books, the frequency of using Shuigou (GV 26) was 240 times, and there were 109 acupoints used in pairs with it.Conclusion The top 10 acupoints used in pairs with Shuigou (GV 26) were Hegu (LI 4), Baihui (GV 20), Zusanli (ST 36), Shangxing (GB 23), Chengjiang (CV 24), Qihai (CV 6), Jiache (ST 6), Fengfu (GV 16), Zhongwan (CV 12), and Weizhong (BL 40). The common depth for acupuncture was 0.4 cun and the dose of moxibu- stion was 3 cones.
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Objective To summarize the rules in the application of Shuigou (GV 26) by sorting the ancient literatures involving Shuigou (GV 26).Method A database was established based on systemically rigidly sorting out the indications, paired acupoints, indications of paired acupoints, acupuncture and moxibustion methods, and stimulation dose of Shuigou (GV 26) by retrieving the 1000 books indexed byZhong Hua Yi Dian (Chinese Canon of Medicine) (4th edition).Result A total of 425 items about the indications of Shuigou (GV 26) were concluded, involving 171 ancient books, the frequency of using Shuigou (GV 26) was 240 times, and there were 109 acupoints used in pairs with it.Conclusion The top 10 acupoints used in pairs with Shuigou (GV 26) were Hegu (LI 4), Baihui (GV 20), Zusanli (ST 36), Shangxing (GB 23), Chengjiang (CV 24), Qihai (CV 6), Jiache (ST 6), Fengfu (GV 16), Zhongwan (CV 12), and Weizhong (BL 40). The common depth for acupuncture was 0.4 cun and the dose of moxibu- stion was 3 cones.