| ObjectiveStarting from the Trigeminal neuro vascular theory of the pathogenesis of migraine,the experimental rat model of nitroglycerin-type migraine was studied and the intervention was carried out using“Hejiezhitong prescription”.To observe the changes of pain-related factors,neuropeptides and Histopathology in plasma,midbrain and trigeminal ganglia of rats,and to explore the mechanism of multi-target action for treating migraine with "palliative analgesics".Methods1.To observe the effect of "Hejiezhitong prescription" on the behavioral characteristics of experimental migraine rats: 48 healthy Wistar rats,half male and half female,were randomly divided into 6 groups.Each group of 8: blank control group(A),migraine model group(B),Oriting control group(C),"Hejiezhitong prescription" high dose group(D),medium dose group(E),low dose group(F).Groups A and B were given saline lavage,and groups D,E,and F were given high,medium,and low doses of "reconciliation analgesics" lavage,once a day for 14 consecutive days;On the 14 th day after filling the stomach,rats in groups B,C,D,E,and F were given subcutaneous injection of nitroglycerin,and group A subcutaneous injection of saline;Group C injected nitroglycerin immediately gave Ole to stop filling the stomach.After 1 hour,he began to observe behavioral changes: scratching the head,biting the tail,climbing the cage,and returning symptoms.He observed for half an hour in a row,and scored and statistically analyzed the symptoms.2.To observe the effects of "Hejiezhitong prescription" on CGRP,5-HT,β-EP,SP,CCK in plasma of migraine rats: 2 hours after molding,the right heart and ear blood were collected.The content of CGRP,5-HT,β-EP,SP,CCK in plasma was detected by ELISA method,and statistical analysis was conducted.3.To observe the effects of "Hejiezhitong prescription" on CGRP,SP,and CCKmRNA in the midbrain and trigeminal ganglion of migraine rats: 2 hours after moulding,the middlebrain and trigeminal ganglion were taken in experimental rats.The expression of CGRP,SP and CCK mRNA in the middle brain and trigeminal ganglion were detected by PCR and statistical analysis was performed.4.To observe the effects of "Hejiezhitong prescription" on the pathology of the midbrain and trigeminal ganglion and the expression of CB1 receptor and COX-2 in migraine rats: 2 hours after moulding,take the middle brain and trigeminal ganglion of rats,make pathological sections,and perform HE staining.Morphological changes of middle brain and trigeminal ganglion were observed under light microscope.The expression of CB1 receptor and COX-2 protein in rat midbrain and trigeminal ganglion specimens was observed by immunohistochemical methods and statistical analysis was carried out.Results1.A model and behavioral observation of nitroglycerin experimental migraine rats(1)After injecting nitroglycerin,the rats in each group showed frequent forelimb scratching,increased number of climbing cages,and increased back and forth movement.The score of migraine model group was higher than that of blank control group,and there were significant differences.(2)Behavioral changes in the treatment group of "Hejiezhitong prescription" and Ole’s treatment group were significantly reduced compared with those of the migraine model group,indicating that each dose group of "Hejiezhitong prescription" could reduce migraine symptoms in rats.The effect is comparable to that of Ole’s treatment group.Compared with the blank control group,there was a significant difference between the low dose of the "Hejiezhitong prescription" and the blank control group,suggesting that the high and medium dose of the "Hejiezhitong prescription" were more effective in improving the behavioral symptoms of experimental rats.2.Effect of "Hejiezhitong prescription" on CGRP,5-HT,β-EP,SP and CCK in plasma of migraine rats(1)The plasma content of CGRP was significantly higher in the migraine model group than that in the blank control group,and the plasma content of CGRP in the high,mediumand low dose of the "Hejiezhitong prescription" group was significantly reduced,which was significantly different from that of the migraine model group.(2)The plasma content of 5-HT in the migraine model group was significantly lower than that of the blank control group,and the plasma content of 5-HT in the high dose group of "Hejiezhitong prescription" was significantly higher,which was significantly different from that of the migraine model group.(3)The plasma content of β-EP was significantly lower than that of the blank control group in the migraine model group,and there was no significant difference between the plasma content of β-EP and the migraine model group in the high,medium and low dose of the "Hejiezhitong prescription" group.(4)There was no significant difference between the SP content in the rat plasma of migraine model group and that of the migraine model group.(5)There was no significant difference in the plasma CCK content of migraine rats.3.Effect of "Hejiezhitong prescription" on expression of CGRP,SP and CCK mRNA in the middle brain and trigeminal ganglion of experimental migraine rats(1)The expression of CGRP mRNA was significantly higher in the migraine model group than in the blank control group.The high,medium and low dose treatment group of "Hejiezhitong prescription" was significantly lower than that of the migraine model group,with significant differences.(2)The expression of SP mRNA was significantly higher in migraine model rats than in blank control group.The high,medium and low dose treatment group of "Hejiezhitong prescription" was significantly lower than that of the migraine model group,with significant differences.(3)The expression of CCK mRNA in middle brain of migraine model rats was significantly higher than that in blank control group.The high,medium and low dose treatment group of "Hejiezhitong prescription" was significantly lower than that of the migraine model group,with significant differences.(4)The expression of CGRP mRNA in the trigeminal ganglion of migraine model ratswas significantly higher than that in the blank control group.There were significant differences between the high,medium and low dose treatment group and the migraine model group.(5)The expression of CCK mRNA in the trigeminal ganglion of migraine rats was significantly higher than that in the blank control group.The high-dose treatment group of "Hejiezhitong prescription" was significantly lower than the migraine model group,with a very significant difference.The low-dose group was significantly lower than the migraine model group,and there was a significant difference.Although the medium-dose treatment group had no significant difference compared with the migraine model group,However,there is a downward trend in the average number compared to the migraine model group.4.Effect of "Hejiezhitong prescription" on middle brain and trigeminal ganglion pathology and expression of CB1 receptor and COX-2 protein in experimental migraine rats.(1)HE staining of midbrain and trigeminal ganglion in migraine model rats was observed under light microscope: there were no obvious abnormalities in the morphology of midbrain neuron cells,trigeminal ganglion cells,and nerve fibers in migraine model rats,and there was no significant change compared with the blank control group.There was no significant change between the high,medium and low dose treatment group and the migraine model group.(2)Immune histochemical method to detect the expression of CB1 receptor in the middle brain and trigeminal ganglion of experimental migraine rats.There was no significant difference between the migraine model group and the blank control group under light microscope.The high,medium and low dose of the "Hejiezhitong prescription" group was significantly stronger than the blank control group and the nitroglycerin model group,with significant differences.(3)The immunohistochemical method for the detection of COX-2 expression in the middle brain and trigeminal ganglion of experimental migraine rats.The migraine model group was significantly stronger than the blank control group,with very significantdifferences.The high,medium and low dose group of "Hejiezhitong prescription" was significantly lower than the migraine model group,with significant differences.The effect of high and medium dose on decreasing the expression of the trigeminal ganglion COX-2was better.Conclusion1.The "Hejiezhitong prescription" can effectively improve the behavioral symptoms of nitroglycerin type migraine experimental rats,and there is no significant difference with Oriting effect,and the high and medium dose effects are better.2.The high,medium,and low doses of "Hejiezhitong prescription" can reduce the CGRP content in rat plasma,and the high dose can increase the content of 5-HT in rat plasma,thereby improving migraine symptoms.3.The "Hejiezhitong prescription" can improve migraine symptoms by reducing the expression of CGRP,SP,and CCK mRNA in the middle brain of rats,affecting the function of the endogenous pain modulation system.4.The "Hejiezhitong prescription" can improve migraine symptoms by reducing the expression of CGRP and CCK mRNA in the trigeminal ganglion of rats.5.The "Hejiezhitong prescription" can activate the expression of CB1 receptors in the middle brain and trigeminal ganglion in rats,reduce COX-2 protein expression,and thus reduce neurogenic inflammatory reactions to play a role in the treatment of migraines. |
PDF Full Text Request