Résumé
Objective To explore the mechanism of the treatment of atrial fibrillation(AF)with traditional Chinese medicine compound of eliminating phlegm and removing blood stasis.Methods Sixty male SD rats were randomly divided into the blank group and the model group.Ach(66 μg·mL-1)-CaCl2(10 mg·mL-1)was injected into the tail vein for 7 consecutive days to establish the rat AF model.The rats with successful modeling were randomly divided into model group,high,medium and low dose groups of Chinese herbal compound and verapamil group.The high,medium and low dose groups of Chinese herbal compound were given 12.38 mg,6.18 mg and 3.10 mg·kg-1·d-1 Chinese herbal compound for removing phlegm and removing blood stasis solution by gavage,while the verapamil group was given 8.31 mg·kg-1·d-1 verapamil solution by gavage,and the blank group and model group were given equal volume distilled water by gavage.During this period,the rats were still given tail vein injection for 14 consecutive days.The duration of atrial fibrillation in lead Ⅱ of rats was measured by electrophysiological recorder,the ultrastructural changes of rat atrial muscle were observed by transmission electron microscope,the relative expression of CAV1.2,CaM,CaMKⅡ mRNA in rat atrial muscle was detected by RT-PCR,and the expression of CAV1.2,CaM,CaMKⅡ and downstream proteins RyR2,P-RyR2 in rat atrial muscle was detected by Western blot.Results Compared with the blank group,the rats in the model group showed typical atrial fibrillation ECG.Compared with the model group,the duration of atrial fibrillation in the compound Chinese medicine group decreased.The arrangement of myofilaments was relatively neat,and the structure of mitochondria was relatively complete;CAV1.2 mRNA and protein expression increased(P<0.05),CaM,CaMKⅡ mRNA and protein expression decreased(P<0.01),downstream protein P-RyR2 expression decreased(P<0.01),RyR2 protein expression had no difference(P>0.05).Conclusion The Chinese herbal compound for removing phlegm and removing blood stasis can shorten the duration of atrial fibrillation in rats,inhibit the ultrastructure damage of atrial myocytes,and its mechanism may be related to regulating the expression of CAV1.2/CaM/CaMKⅡ signal pathway and improving the disorder of calcium regulation.
Résumé
Objective To investigate the role of CaM/CaMK-Ⅱ signaling pathways in inflammatory pain in mice.Methods Sixty male C57BL6 mice,weighing 25-27 g,were randomly divided into 3 groups (n =20):control group (group C),complete freunds adjuvant (CFA) group (group F) and KN-93+CFA group (group KF).Saline 50 μl were injected into the right side of the claw in group C.CFA 50 μl were injected into the right claw foot for the preparation of inflammatory pain models in group F.KN-93 45 nmol was injected i.c.v.30 min before CFA injection in group KF.The thermal withdrawal latency (TWL) were measured 30 min before injection,1 h and 4 h after injection.The protein expressions of CaMK-Ⅱ,c-fos and CREB in the spinal cord were measured at above time by Western blot.Results Compared with group C,TWL were lower in groups F and KF 1 h and 4 h after injection (P<0.05).Compared with groups F,TWL in group KF were higher 1 h and 4 h after injection (P<0.05).Compared with group C,the protein expressions of p-CaMK-Ⅱ,p-CREB,e-fos and mRNA expression of CaMK-Ⅱ,CREB,c-fos were higher in group F and KF 1 h and 4 h after injection (P<0.05).Compared with group F,the protein expression of p-CaMK-Ⅱ,p-CREB,c-fos and mRNA expressions of CaMK-Ⅱ,CREB,c-fos in group KF were lower 1 h and 4 h after injection (P<0.05).Conclusion CaM/CaMK-Ⅱ signaling pathways involved in inflammatory pain in mice.