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.
ABSTRACT
Background & Objective: The calcitonin gene-related peptide (CGRP) has a central role in the pathogenesis of migraine, but variations in CGRP-related genes, including the calcitonin gene-related polypeptide-alpha (CALCA) gene and the receptor activity modifying 1 (RAMP1) gene, have not been found to link with migraine in Australian population. The goals of this study were to determine whether variants in the two genes are related to migraine in Chinese population. Methods: Using a case-control approach, rs3781719 and rs145837941 in the CALCA gene and rs3754701 and rs7590387 at the RAMP1 locus was analyzed in a cohort of 504 migraine cases and 529 ethnically matched controls. Genotyping was performed using Sequenom MALDI-TOF mass spectrometry iPLEX platform. Results: The CALCA gene rs145837941 variant was not found in migraine or control group. No significant difference in genotypic and allelic distribution was observed in the other three polymorphisms between migraine cases and controls. All the three SNPs were also not selected as significant factors that independently contributed to susceptibility to migraine in multivariate analysis. In the subgroup analysis, the CALCA rs3781719 seemed to be a significant risk for migraine with aura, but was not statistically significant after FDR correction. Moreover, there was no synergistic relationship between the three SNPs in the multifactor dimensionality reduction analysis for explore locus–locus interactions. Conclusion: Our data suggested that variants in CALCA gene and RAMP1 gene were not associated with migraine in the Han-Chinese population.
Subject(s)
Calcitonin Gene-Related Peptide , Migraine DisordersABSTRACT
Objective:To investigate the effect of blocking the expression of receptor activity modifying protein 1 (RAMP1 )on calcito-nin gene-related peptide(CGRP)-induced MG-63 cell proliferation.Methods:RAMP1 siRNA was synthesized and screened by tran-scription in vitro.The subcultured MG-63 cells were divided into the following groups:RAMP1 siRNA interference group,empty vector group and blank control group.The mRNA expression and the membrane distribution changes of the calcitonin receptor-like receptor (CRLR)and the receptor component protein (RCP)in MG-63 cells were examined by real-time PCR and immunofluorescence method respectively.Results:RAMP1 and CRLR mRNA and the fluorescence intensity of MG-63 cells decreased after transfection by RAMP1 siRNA(P <0.05).In RAMP1 interference group,the expression of RCP mRNA and the fluorescence intensity were higher than those in the other two groups(P <0.05).After RAMP1 siRNA interference,the proliferation of MG-63 cells was inhibited(P <0.05). Conclusion:RAMP1 siRNA transfection may reduce CRLR expression and inhibite the proliferation of MG-63 cell.