An Experimental Research On The Degranulation Of Mast Cells In The Dura Mater After The Intervention Of Yangxueqingnao Granules

Objectives:This study mainly appraised the behavior of NTG-induced migraine rats and detected the degranulation rate of mast cells in their dura mater after the intervention of Yangxueqingnao Granules, to explore its function in the pathogenesis of migraine and to investigate the experimental therapeutic action of Yangxueqingnao Granules on migraine rats.Methods:1. Healthy male Wistar rats were randomly divided into normal saline control group, NTG model group, Yangxueqingnao Granules high-dose-prevention group, Yangxueqingnao Granules low-dose-prevention group, Yangxueqingnao Granules high-dose-treatment group, Yangxueqingnao Granules low-dose-treatment group, and18rats were included in each group. Each was further divided into30min、60min and180min group, in which groups the rats would be killed at the corresponding time point after modeling.2. The migraine model was established by injecting NTG subcutaneous. The normal saline control group received the same volume of saline.3. Yangxueqingnao Granules high-dose (or low-dose)-treatment group was given high-dosage (or low) gavage of Yangxueqingnao Granules10min after modeling, while Yangxueqingnao Granules high-dose (or low-dose)-prevention group was given high-dosage (or low) of Yangxueqingnao Granules for7days by intragastric administration before modeling. 4. We observed the migraine relevant behavior, such as scratching head, red ears and climbing hutch, recorded the time when these behavior appeared or peaked or declined, acquired the occurrence frequency and duration. The migraine behavior scores of each group were continuously recorded at an interval of30minutes after modeling, according to the behavioral scoring standard.5. After the rats were killed, the dura matter was gained and specially stained using toluidine blue. The degranulation rate of mast cell in the dura was then studied. The level of significance was0.05.Results:1. Behavior scores: Compared with the normal saline control group, the scores of the treatment group were significantly higher(P<0.05), but there was no significant difference between the treatment group and the NTG model group(P>0.05). And compared with the NTG model group, the scores of the Yangxueqingnao Granules prevention group were significantly lower within30min after modeling(P<0.001), while there was no significant difference when compared with the normal saline control group (P>0.05), and the score of the normal saline group was higher during this period than any other periods. During30min to150min after modeling, the behavior scores showed a significant difference among the normal saline group, Yangxueqingnao Granules prevention group and the treatment group(P<0.001), and the difference between the Yangxueqingnao Granules high-dose-prevention group and the low-dose-prevention group was significant during90-120min. Analysis of variance of factorial design showed that the sequence of administration (prevention or treatment) influenced the behavior score more than the dosage and their interaction.2. Degranulation rate: Toluidine blue staining indicated that the degranulation rate in normal saline group was significantly lower than any other groups at each time point (P<0.05), and that fewer dural mast cells became degranulated in Yangxueqingnao Granules prevention groups than NTG model group(P<0.05), while there was no obvious difference between high-dose group and low-dose group(P>0.05). However, the dural mast cell degranulation in Yangxueqingnao Granules treatment group showed no serious difference from NTG model group (P>0.05). Analysis of variance of factorial design showed that the sequence of administration (prevention or treatment) influenced the degranulation rate more than the dosage and their interaction.Conclusion:1. NTG-induced migraine model in rat has certain similarities with that in human migraine, and NTG can also induce dural MC degranulation.2. Prophylactic use of Yangxueqingnao Granules significantly reduced the behavior score and MC degranulation rate of migraine rats, prompting that it has a superior preventive effect on migraine, and that it may contribute as the MC stabilizing agent.3. However, the Dose-effect relationship between Yangxueqingnao Granules and migraine still need further study.