| Objective:To observe the activity of morphology and structure of rat mitochondria and the mitochondrial respiratory chain complex Ⅰ-Ⅴ element in migraine rats,and to provide a theoretical basis for the prevention and treatment of migraine by acupuncture.Methods:We choose healthy male SD rats with body weight of 200 ～ 250 g,which were divided into blank group,sham operation group,model group(3 groups were no intervention group),western medicine pretreatment group,acupuncture pretreatment group,acupuncture plus western medicine(pretreatment group),We have 8 rats in each group,the grouping method according to random number table method.Acupuncture pretreatment group and was treated with electroacupuncture intervention(density wave,2/15 Hz,1m A,20min/ once,once a day)at Taichong and Yanglingquan acupoints 14 days before modeling.Western medicine group was given intraperitoneal injection of flunarizine hydrochloride capsule(20mg/kg,once a day)on the 14 th day of modeling.Each group was modeled after 14 days of intervention(CSD model),and cervical vein blood was collected immediately after non-invasive blood flow test,and brain was taken after severed head.The morphology and structure of mitochondria in the frontal cortex of CSD model rats were observed by transmission electron microscopy.The activity of the mitochondrial respiratory chain complex Ⅰ-Ⅴ was determined by enzymological method.The expression of c-fos in the frontal cortex and the trigeminal spinal tract nucleus was measured by immunofluorescence method,and the content of no in rat serum was determined by microplate method。The expression of c-fos in the frontal cortex and the trigeminal spinal tract nucleus 、he content of no in rat serum are effect indicators.Results:1 Transmission electron microscopy showed that the morphology and structure of mitochondria in the affected side of the cortex of rats in the model group were moderate to severe swelling,forming vacuoles,suggesting that the function was reduced or disappeared,and the quantity percentage of abnormal mitochondria in model group was noticeably higher than the data in the blank group and sham group(P < 0.05),while compared blank group with sham group,P>0.05.Compared with model group,the abnormal percentage of acupuncture and western medicine pretreatment group were lower than it(P < 0.05),while compared with blank group,P>0.05.2 Improvement of activity of Ⅰ-Ⅴ of cortical mitochondrial respiratory chain complex in migraine model rats by acupuncture preconditioning(1)Results of The activity of mitochondrial complex Ⅰ :The activity in model group was much lower than the data in blank group and sham group(P < 0.05),while compared blank group with sham group,P< 0.05.Compared with model group,the activity of acupuncture and western medicine pretreatment group showed obvious increased trend(P < 0.05),while compared with blank group,P>0.05.There was no obviously difference in the activity of complex I between the acupuncture and western medicine pretreatment group(P > 0.05).(2)Results of the activity of mitochondrial complex Ⅱ :Compared to model group,the activity in blank group was higher than the activity of model group,but the variance was not obviously significant(P > 0.05).The activity of mitochondrial complex Ⅱ in acupuncture and western medicine pretreatment group were obviously higher than the data in model group,and the dissimilarity was statistically significant(P < 0.05),while compared to the blank group,P > 0.05.There was no obviously difference in the activity of complex Ⅱ between the acupuncture and western medicine pretreatment group(P > 0.05).(3)Results of the activity of mitochondrial complex Ⅲ :The activity of the model group was lower than the data of the sham operation group and the blank group,but the dissimilarity was not obviously different(P > 0.05).Compared to model group,the activity between the acupuncture and western medicine pretreatment group had significant trend of increasing,but the dissimilarity was not obviously different(P >0.05).There was no obviously difference in the activity of complex Ⅲ between the acupuncture and western medicine pretreatment group(P > 0.05).(4)Results of the activity of mitochondrial complex Ⅳ: The activity in model group was much lower than the data in blank group(P < 0.05),but compared to sham operation group,P > 0.05,while compared blank group with sham operation group,P< 0.05.Compared to model group,the activity of the intervention groups showed an increasing trend,P < 0.05.Compared with the activity of the acupuncture group and the western medicine group was significantly increased,the dissimilarity was not obviously different(P > 0.05).(5)Results of the activity of mitochondrial complex Ⅴ: The activity in model group was much lower than the data in blank group and sham operation group(P < 0.05),while compared blank group with sham group,P< 0.05.Compared with model group,the activity of complex Ⅴ in acupuncture and western medicine pretreatment group showed obvious increased trend(P < 0.05),while compared with blank group,P>0.05.There was no obviously difference in the activity of complex Ⅴ between the acupuncture and western medicine pretreatment group(P > 0.05).3 Acupuncture preconditioning improved the expression of c-fos in cortex and trigeminal spinal tract nucleus and the concentration of NO in serum of migraine model rats(1)Results of serum NO content: The content of model group was obviously higher than the data in the blank group and sham operation group(P < 0.05),while compared blank group with sham operation group,P > 0.05.Compared with the model group,the content of acupuncture and western medicine pretreatment group were obviously decreased(P < 0.05),while compared the content of the intervention groups wihe the blank group,P > 0.05.There was no obviously difference in the content of NO between the acupuncture and western medicine pretreatment group(P > 0.05).(2)Results of C-fos expression positive cell count: The C-fos expression positive cell count in the cortex and trigeminal spinal nucleus of rats in model group was much higher than that in sham operation group and blank group(P < 0.05),while compared blank group with sham operation group,P > 0.05.Compared to the model group,the positive cell count of C-fos expression was obviously decreased in the acupuncture and western medicine pretreatment group(P < 0.05),while compared intervention groups to blank group,P > 0.05.There was no obviously difference in the C-fos expression positive cell count between the acupuncture and western medicine pretreatment group(P > 0.05).Conclusion:1)Acupuncture and Western Medicine(flunarizine hydrochloride)pretreatment can prevent the emergence of CSD.2)The mechanism of acupuncture and Western Medicine(flunarizine hydrochloride)in preventing the emergence of CSD may be through protecting mitochondrial structure and improve the activity of mitochondrial respiratory chain complex I-V,thus reducing the expression of c-fos in cortex and spinal trigeminal nucleus and the content of no in serum of CSD model rats. |
PDF Full Text Request