| Objective: To observe the effect of Lu Tong Granul on the behavior of migraine rat and the expression of NF-κB,P-IκBα,IL-1β,TNF-a and COX-2 protein,and to explore the possible mechanism.Methods: 1.Grouping: 60 healthy male rats were randomly divided into normal group,model group,sumatriptan group,high-dose group of Lu Tong Granul(short-term high-dose group),middle-dose group of Lu Tong Granul(short-term middle-dose group),low-dose group of Lu Tong Granul(short-term low-dose group),10 rats in each group.2.Dosing: The high,medium,and low dose groups of Lu Tong Granul were given intragastrically at doses of 4g,2g,1g/kg/d,and the sumatriptan dose was 6mg/kg/d.normal group and model group were given equal volume of distilled water,gavage once a day for 7 days.3.Modeling: The migraine model was replicated after the last intragastric administration for 1 hour.Rats in the normal group were subcutaneously injected with 10mg/kg of back physiologic saline,and the rest of the rats were subcutaneously injected with 10mg/kg of nitroglycerin(NTG).4.Detection indicators:(1)Behavioral Change of rats: Observe and record the occurrence and disappearance of ear-redness in 60 rats within 180 min after successful replication of the migraine model.At the same time,every 30 minutes was used as a time period to record the number of scratching heads and climbing Cages in 60 rats within 180 minutes.(2)NF-κB p65,P-IκBα protein expression: 2% sodium pentobarbital 50mg/kg abdominal anaesthesia in rats,each group randomly selected 5 rats decapitated fresh brain tissue,The tissues were placed on ice,referring to the rat brain stereotaxic map,we isolate the brain stems.The brain stems were placed in an EP tube and stored in a refrigerator at-80°C.and used for Western Blot to detect the expression of NF-κB p65 and P-IκBα protein(3)Brain stem IL-1β,TNF-a,and COX-2 positive cells: Rats were perfused to obtain brains.Brain stems were isolated by referring to the rat brain stereotaxic map,and used for immunohistochemistry to detect the immunoreactive positive cell number of IL-1β,TNF-a,COX-2.Results:1.Behavioral changes: Compared with the model group,the duration of ear redness was shortened in the high-,middle-,and low-dose group of Lu Tong Granul and the sumatriptan group,and the number of scratching heads and climbing cages was reduced.The difference was statistically significant(P<0.05).Compared with the low dose group,the duration of ear redness was shortened and the number of scratching heads and climbing cages was reduced in the high-,medium-,and the sumatriptan group.The difference was statistically significant(P<0.05).Compared with the middle-dose group,the duration of ear redness was shortened in the high-dose group of Lu Tong Granul and the sumatriptan group,and the number of scratching heads and climbing cages was reduced,the difference was statistically significant(P<0.05).Compared with the sumatriptan group,although the ear redness time was shortened and the number of disturbing heads and climbing cages was reduced in the high-dose group of Lu Tong Granul,there was no statistically significant difference(P>0.05).2.The NF-κB p65 content of Brain stem: Compared with the normal group: The expression level of NF-κB p65 protein in the brainstem of rats in the model group and each administration group was increased(P<0.05).Compared with the model group,the expression level of NF-κB p65 protein in the brainstem of rats with high,middle,and low-dose group of Lu Tong Granul and the sumatriptan group was reduced,the difference was statistically significant(P<0.05).Compared with the low dose group,the expression level of NF-κB p65 protein in the brainstem of rats in the middle,high-dose group of Lu Tong Granul and the sumatriptan group was reduced,the difference was statistically significant(P<0.05).Compared with the middle-dose group,the expression level of NF-κB p65 protein in the brainstem of rats in the high-dose group of Lu Tong Granul and the sumatriptan group was decreased,and the most significant decrease was observed in the high-dose group(P<0.05).There was no significant difference in the high-dose group of Lu Tong Granul and the sumatriptan group(P>0.05).3.The P-IκBα content of Brain stem: Compared with the normal group,the expression level of P-IκBα protein in the brainstem of rats in the model group and each administration group was increased(P<0.05).Compared with the model group,the expression level of P-IκBα protein in the brainstem of rats in the high,middle,and low-dose group of Lu Tong Granul and the sumatriptan group was reduced,the difference was statistically significant(P<0.05).Compared with the low dose group,the expression level of P-IκBα protein in the brainstem of rats in the middle,high-dose group of Lu Tong Granul and the Sumatriptan group was reduced,the difference was statistically significant(P<0.05).Compared with the middle-dose group,the expression level of P-IκBα protein in the brainstem of rats in the high-dose group of Lu Tong Granul and the sumatriptan group was decreased,and the most significant decrease was observed in the high-dose group(P<0.05).There was no significant difference in the high-dose group of Lu Tong Granul and the sumatriptan group(P>0.05).4.The expression of IL-1β,TNF-a,and COX-2 positive cells in brain stem: Compared with normal group,the number of positive cells of IL-1β,TNF-a,and COX-2 in brainstem of rats in the model group and each administration group was increased(P<0.05).Compared with the model group,the number of positive cells of IL-1β,TNF-a,and COX-2 in the brainstem of rats in the high,middle,and low-dose group of Lu Tong Granul and the sumatriptan group was reduced,the difference was statistically significant(P<0.05).Compared with the low dose group,the number of positive cells of IL-1β,TNF-a,and COX-2 in the brainstem of rats in the middle,high-dose group of Lu Tong Granul and the Sumatriptan group was reduced,the difference was statistically significant(P<0.05).Compared with the middle dose group,the number of positive cells of IL-1β,TNF-a,and COX-2 in the brain stem of rats in the high-dose group of Lu Tong Granul and the sumatriptan group was decreased,and the most significant decrease was observed in the high-dose group(P<0.05).There was no significant difference in the high-dose group of Lu Tong Granul and the sumatriptan group(p>0.05).Conclusion: 1.Lu Tong Granul can shorten the time of ear redness in migraine model rat,reduce the number of scratching heads and climbing cages,and have the effect of treating migraine.2.Lu Tong Granul can reduce the expression of NF-κB p65,P-IκBα,IL-1β,TNF-a and COX-2 proteins in brain stem of migraine model rat.3.The activation of NF-κB signaling pathway is involved in the pathogenesis of migraine,and the mechanism of Lu Tong Granul may be related to the inhibition of the activation of NF-κB signaling pathway.4.Lu Tong Granul has a dose-dependent effect on migraine model rat,with the best dosage is high dose. |
PDF Full Text Request