ABSTRACT
OBJECTIVE To expl ore the mechanism of effe ctive fractions from Xiongma decoction in the treatment of migraine with hyperactivity of liver-yang and blood stasis. METHODS Totally 70 male SD rats were randomly divided into normal group ， model group ，positive control group （Flunarizine hydrochloride capsules 0.9 mg/kg），low-dose and high-dose groups of Xiongma decoction effective fractions （ethyl acetate extract 0.87，3.46 g/kg，n-butanol extract 1.80，7.20 g/kg）. Except for normal group ， rats in other groups were given aconite decoction （2 g/kg），once a day ，for 4 consecutive weeks to establish the hyperactivity model of liver-yang. On the 15th day of modeling ，all administration groups were given corresponding drugs intragastrically at the same time ，once a day ，for 2 consecutive weeks. On the 29th day of modeling ，rats trigeminal ganglion was stimulated to establish the migraine model with hyperactivity of liver-yang and blood stasis ，then the medication was maintained for another time according to the above method. The macroscopic signs and behavior of the rats were observed ；positive expression ，mRNA and protein expression of transient receptor potential vanilloid 1 （TRPV1），calcitonin generelated peptide （CGRP），calcitonin receptor-like receptor （CRLR） and receptor-associated membrane proteins 1 （RAMP1） in trigeminal cervical spinal complex （TCC） were detected by immunohistochemistry ，RT-qPCR and Western blot assay. RESULTS Rats in model group showed macrophysical signs and behaviora l manifestations related to migraine with hyperactivity of liver-yang and blood stasis. Thirty minutes after last administration ，the above conditions of rats in Xiongma decoction effective fraction groups were improved significantly. Compared with normal group ， positive expression，mRNA and protein expression of TRPV 1，CGRP， CRLR and RAMP 1 in TCC of rats in the model group were significantly increased （P＜0.05）. Compared with model zengguirong@hnse.org group， most of above indicators in Xiongma decoction effective fraction groups were significantly decreased （P＜0.05）. CONCLUSIONS The mechanism of Xiongma decoction in preventing and treating migraine with hyperactivity of liver-yang and blood stasis may be related to inhibit the activity of TRPV1-CGRP/CGRP receptor signaling pathway in TCC.
ABSTRACT
Objective To explore the effects of bioactive parts of Xiongma Decoction (parts of ethyl acetate and n-butanol extract) on the CGRP-CRLR/RAMP1 signal pathway so as to clarify its therapeutic mechanism on migraine.Methods We randomly divided 36 male SD rats into 6 groups with 6 in each:blank group,model group,groups of low-,medium-and high-dose Xiongma Decoction bioactive parts,and Sumatriptan group.By giving hypodermic injection of 10 mg/kg nitroglycerin,the migraine rat model was copied;Only 18 rats were found to have positive expressions of CGRP,CRLR,and RAMP1 in TCC with immunohistochemistry staining after heart perfusion.For the remaining 18 rats,TCC was stripped directly from the whole brain and divided into two parts,one part used to detect CGRP,CRLR,RAMP1 mRNA expressions by qPCR,and the other part to detect CGRP,CRLR,RAMP1 protein expressions by Western blot.Results The number of CGRP,CRLR and RAMP1 immunoreactive cells,the mRNA and protein expressions on TCC in model group were effectively increased,compared with those in the blank group (P＜0.05),indicating that the model copying was successful.Compared with those in the model group,the number of CGRP,CRLR and RAMP1 immunoreactive cells in Xiongma Decoction bioactive parts was significantlv decreased,and the expressions of CGRP,CRLR and RAMP1 mRNA and protein were reduced (P＜0.05).Conclusion The bioactive parts of Xiongma Decoction can reduce the activity of CGRP-CRLR/RAMP1 signal pathway in TCC of migraine rats.