| Objective:On the basis of the effectiveness of Huayutongluo Moxibustion in the treatment of VD,to see how Huayutongluo Moxibustion affect the expression of Sonic Hedgehog(Shh)signal pathway components in the corpus callosum of VD rats,as well as the expression of myelin regeneration-related proteins PLP and NG2,so as to explore the mechanism of the effect of Huayutongluo Moxibustion on promoting myelin regeneration in VD rats.Method:Experiment 1: SPF male Wistar rats were selected to replicate VD model by BCCAO method.With 12 rats in each group,36 animals successfully replicated in the model were randomly divided into four groups: model,moxibustion,western medicine,and sham operation as a blank control.Huayutongluo Moxibustion was used on the moxibustion group.The course of treatment was once a day,over the course of seven days,three points of suspended moxibustion "Shenting","Baihui" and "Dazhui" were administered for 20 minutes,after each session of therapy,everyone rested for one day,for a total of three courses of treatment.The western medicine group received Clemastine solution intragastric administration twice a day,with the same treatment schedule as the moxibustion group.The model and sham operation groups received no treatment.and were only captured and placed on the self-made moxibustion rack during the treatment time parallel to the moxibustion group and the western medicine group.Morris water maze experiment was carried out after the treatment course,the changes in learning ability and memory of the rats in each group were also compared and analysed within and between groups to confirm the improvement in learning ability and memory of VD rats.Experiment 2: Real-time quantitative PCR(q RT-PCR)and Western blotting(WB)were used to detect the expression of Shh signal pathway components Shh and Gli 1 in the corpus callosum of rats.NG2 was used to mark the proliferation of OPCs and PLP was used to mark the formation of m OL.The evidence of moxibustion promoting myelin regeneration in VD rats was obtained at protein level.Results:1.Effects of Huayutongluo Moxibustion on learning and memory ability in rats with VDPosition navigation experiment:(1)Pattern recognition(before intervention):Compared to the dummy group,the escape latency of rats in the dummy group was significantly longer(P<0.05),indicating that the pattern was successful.(2)Postintervention: The escape latency in the model group was always longer than that in the sham operation group(P<0.01);the escape latency in the moxibustion and western medicine groups was considerably lower than in the model group(P<0.01).There was no statistically significant difference between the moxibustion and western medicine groups(P>0.05).2.Effect of Huayutongluo Moxibustion on the expression of Shh signal pathway components in each group of ratsWestern Blot analysis revealed that the expression of Shh and Gli1 was lower in the model group than in the sham surgery operation group(P<0.05).Shh and Gli1 expression was higher in the moxibustion and western medicine groups compared to the model group(P<0.01).There was no statistically significant difference between the moxibustion and western medicine groups(P>0.05).The q RT-PCR findings revealed that Shh and Gli1 m RNA expression levels were lower in the model group than in the sham operation group(P<0.01).Shh and Gli1 m RNA expression levels were greater in the moxibustion and western medicine groups than in the model group(P<0.01).The moxibustion and western medicine groups did not vary significantly(P>0.05).Huayutongluo Moxibustion can improve VD rats’ learning and memory abilities,as well as increase the expression of Shh signaling pathway components and myelin regeneration associated protein PLPNG2 in the corpus callosum area.3.Effect of Huayutongluo Moxibustion on the expression of PLP and NG2 proteins in each groupCompared with the sham operation group,the average fluorescence intensity of PLP and NG2 decreased in the model group(P<0.01),while the average fluorescence intensity of PLP and NG2 increased significantly in the moxibustion and western medicine groups(P<0.01).There was no significant difference between the western medicine group and the moxibustion group(P>0.05).Conclusion:Huayutongluo Moxibustion can improve VD rats’ learning and memory abilities,as well as increase the expression of Shh signaling pathway components and myelin regeneration associated protein PLP、NG2 in the corpus callosum area,promote the differentiation and maturation of OPC after cerebral ischemia,promote myelin regeneration and repair white matter injury. |
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