| Objective: By observing the changes of the average area percentage of cerebral infarction,the permeability of blood-brain barrier(BBB),the water content of brain tissue,the expression of AQP4 and AQP1 induced by Xingnao Kaiqiao acupuncture therapy,BYHWD,Xingnao Kaiqiao acupuncture therapy combined with BYHWD in MCAO rats,we exploring the possible mechanisms of the treatment of cerebral infarction by combination of acupuncture therapy and medicine.Methods: A total number of 220 SPF grade SD rats,220-250 g,half of male and half of female,were randomly divided into the sham group(group A),the control group(group B),the acupuncture group(group C),the BYHWD group(group D),and the acupuncture+BYHWD combination group(group E).The middle cerebral artery occlusion(MCAO)model was subjected to the group of control,acupuncture,BYHWD and the acupuncture+BYHWD combination.Behavioral scores were performed on the neurological defects of the rats after successful modeling.In the group of the sham,only blunt vessel separation was performed.The group of the sham and the group of acupuncture+BYHWD combination were treated with acupuncture+traditional Chinese medicine,the group of acupuncture was treated with Xingnao kaiqiao acupuncture therapy,the group of BYHWD was oral administrated with BYHW decoction,and the model group was without any treatment.Operation: Xingnao kaiqiao acupuncture therapy was carried out at bilateral Neiguan(PC6),Shuigou(GV26),Sanyinjiao(SP6)on affected side and the needles were remained for 20 min,at twice a day and for 5 days consecutively.The acupoints were referred to Experimental Acupuncture.All rats in the groups were taken at two time points of 3d and 5d,and 6 animals were taken at each time point respectively.The average area percentage of cerebral infarction was determined by TTC staining method,the permeability of BBB was determined by Evans blue staining method,the water content of brain tissue was determined by wet and dry weight method,and the expression of AQP4 and AQP1 was determined by Western Blot method.Results: 1.The average area percentage of cerebral infarction: the overall data showed that the average cerebral infarct area ratio in the group of acupuncture,BYHWD,and acupuncture +BYHWD combination was smaller than that in the model group,and group of acupuncture +BYHWD combination was smaller than that in the group of acupuncture and BYHWD,in which there was no statistically significant difference between the groups at 3d(P>0.05),and the difference between the group of acupuncture + BYHWD combination and acupuncture at 5d was statistically significant(P<0.05).2.Permeability of BBB:(1)3d after surgery,compared with the model group,the Evans blue content in brain tissue gradually decreased in the group of acupuncture,BYHWD and acupuncture + BYHWD combination,and combination group was the lowest,but the difference was not statistically significant(P > 0.05);(2)5d after surgery,compared with the model group,the differences of EB content in brain tissue in the group of acupuncture,BYHWD and acupuncture + BYHWD combination was statistically significant(P< 0.01),the EB content in brain tissue was significantly reduced after treatment with acupuncture,BYHWD,and acupuncture and acupuncture + BYHWD;(3)from 3d to 5d,the EB content in brain tissue generally showed a decreasing trend compared with the model group.3.Water content of brain tissue:(1)3d after surgery,compared with the model group,there was a statistically significant difference only in the group of acupuncture + BYHWD combination(P <0.01);meanwhile there was no significant difference between only the combination group and the sham group(P > 0.05),that is,the water content of brain tissue in combination group was similar to that in sham group.Compared with acupuncture group,the difference in the group of BYHWD and acupuncture + BYHWD combination was statistically significant(P < 0.05),and there was a strongly statistically significant between the combination group and acupuncture group(P < 0.01).That means the water content of brain tissue in the group of BYHWD and acupuncture + BYHWD combination was significantly reduced compared with the acupuncture group.However,the water content of brain tissue in combination group was lower than that in BYHWD group,but the difference was with no statistically significant(P > 0.05).(2)From 3days to 5 days after surgery,the water content of brain tissue generally decreased,with the lowest value in the acupuncture and drug combination group.4.Expression of AQP4 and AQP1:(1)Expression of AQP4:First,compared with B group at 3 and 5 days after surgery,there were statistically significant differences between the group of acupuncture,BYHWD and acupuncture+ BYHWD combination(P < 0.05).The expression of AQP4 in the groups above was lower than that in the model group,and the combination group was the lowest.Secondly,compared with the acupuncture group,there were statistically significant differences between the group of BYHWD and the group of acupuncture + BYHWD combination(P < 0.05),and the difference between the combination group was strongly statistically significant(P < 0.01),and the expression of AQP4 in the combination group was significantly lower than that in the acupuncture group and BYHWD group.Thirdly,there was a decreasing trend in the group of acupuncture + BYHWD combination compared with the BYHWD group,even though there was no statistical significance between them(P > 0.05).(2)AQP1 expression: 3 days after operation,compared with model group,AQP1 expression in all treatment groups showed a decreasing trend,the difference of the acupuncture group was statistically significant(P < 0.05),the difference of the acupuncture + BYHWD combination group showed a enormously statistically significant(P <0.01),and the expression of AQP1 in the combination group was the lowest;Compared with the BYHWD group,the difference was statistically significant(P < 0.05),and the expression of AQP1 in the group of acupuncture + BYHWD combination was lower than that in the BYHWD group.Conclusion: 1.Acupuncture combined with BYWH decoction can inhibit the expression of AQP4 and AQP1,reduce permeability of BBB and improve the edema of ischemic brain tissue to reduce the area of cerebral infarction,and its curative effect is better than that of acupuncture and BYHW decoction alone.2.Combined with the pathological mechanism of BBB destruction and ischemic brain edema at different stages after cerebral infarction,the intervention of acupuncture,BYHW decoction,and the combination of acupuncture and BYHW decoction should be implemented in the early stage of cerebral infarction as soon as possible.3 days after cerebral infarction may be the key time point for treatment,and AQP4 is the key target. |
