| Objective:To observe the effects of Huayu Tongluo moxibustion on myelin-related protein expression and myelin ultrastructure in the corpus callosum region of rats with vascular dementia,and to provide a basis for moxibustion treatment of VD.Mothods:Experiment 1: Wistar rats were screened by Morris water maze and randomly divided into sham operation group,model group,moxibustion group and western medicine group,with 12 rats in each group.Except for the sham-operated group,the VD rat model was prepared by permanent ligation of the bilateral common carotid arteries in all the groups.In the moxibustion group,the rats were suspended from "Baihui","Dazhi" and "Shenting" for 20 min,once a day;in the western medicine group,the rats were gavaged with clomastine solution twice a day;The sham-operated group and the model group were only placed on the moxibustion frame without any other treatment,and the intervention time was the same as that of the moxibustion group.Above four groups 7d for a course of treatment,a total of 3 courses of treatment.After 3 sessions,the Morris water maze experiment was used to observe the changes in learning memory ability of each group of rats before and after the intervention.Experiment 2: The rats were subjected to brain tissue sampling after Morris water maze assay,and the ultrastructure of myelin sheath and the number of myelinated axons in the corpus callosum region of each group were observed by transmission electron microscopy,and the expression of myelin-related proteins MBP,MAG and caspr-Nav1.6-caspr in the corpus callosum region of rats were detected by immunofluorescence and Western blot.Results:(1)The results of the localization navigation experiment: after 3 d of modeling,compared with the sham-operated group,the evasion latency of the VD model group was prolonged(P<0.05),suggesting that the VD rat model was successfully prepared.3courses of treatment later,intra-group comparison: the evasion latency of the moxibustion and western medicine groups was shortened(P<0.05),and there was no significant change between the sham-operated and model groups(P>0.05).Between-group comparison: the escape latency was prolonged in the model group compared with the sham-operated group(P<0.05);the escape latency was shortened in the moxibustion and western medicine groups compared with the model group(P<0.05);there was no statistically significant difference between groups in the escape latency of the moxibustion group compared with the western medicine group(P>0.05).(2)Transmission electron microscopic results: in the sham-operated group,the myelin structure was arranged in a coil-like pattern,forming light and dark semi-layers with regular structure,dense arrangement and abundant myelin sheaths;in the model group,the damage to the myelin structure was more obvious,with different degrees of discrete lamellar structure,varying thickness,widened gaps and sparse myelin sheaths;Compared with model group,the number of myelin sheath in moxibustion group and western medicine group was more than that in model group,and the structure of myelin sheath was repaired.Compared with the sham-operated group,the number of myelinated axons per unit area in the model group decreased(P<0.05);compared with the model group,the number of myelinated axons in the moxibustion and western medicine groups increased(P<0.05);compared with the western medicine group,the number of myelinated axons in the moxibustion group increased(P<0.05).(3)Immunofluorescence and Western blot results: compared with the sham-operated group,the expression levels of MBP,MAG and caspr-Nav1.6-caspr decreased in the model group(P<0.05);compared with the model group,the expression levels of MBP,MAG and caspr-Nav1.6-caspr increased in the moxibustion and Western medicine groups(P<0.05);compared with the Compared with the Western medicine group,the expression levels of MAG and caspr-Nav1.6-caspr increased in the moxibustion group(P<0.05),and the differences between groups in the expression levels of MBP were not statistically significant(P>0.05).Conclusion:Huayu Tongluo moxibustion can improve the learning and memory ability of VD rats and the myelin structure of corpus callosum region,promote the expression of myelin-related proteins MBP,MAG and caspr-Nav1.6-caspr in the corpus callosum region of VD rats,and promote myelin regeneration. |
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