| Objective: Through animal experimental study,to explore the correlation between morphological damage of hippocampal CA1 region of VaD rats and local inflammatory response after cerebral ischemia and hypoxia,to study the mechanism of morphological damage of VaD,to detect the expression of inflammatory factors in hippocampal CA1 region of VaD rats after different intervention treatments,to clarify the effect of scalp acupuncture on inflammatory factors in hippocampal CA1 region of VaD rats,and to explore the mechanism of protecting neurons and improving cognition.Methods: The healthy SD rats were selected to establish the VaD rat model by Olsson method,and the water maze behavior test was used to detect the modeling effect.Then the successful VaD rats were randomly divided into four groups: model group,exercise training group,scalp acupuncture group and Scalp acupuncture exercise acupuncture group and each group has 8animals.In the sham operation group,8 rats only exposed the common carotid artery without bilateral common carotid artery clamping.A total of 5 groups of rats were used for the final experimental study.After the completion of the grouping,the corresponding treatment was carried out according to the group.the sham operation group and the model group were only fed normally without additional treatment;the exercise training group was given exercise rehabilitation training;the scalp acupuncture group was divided into seven areas of Yu’s scalp acupuncture,and the parietal,preparietal and frontal regions(i.e.Shenting,Baihui,Qianding and 5mm on the left and right side of the three acupoints)were given intradermal acupuncture.The scalp acupuncture exercise acupuncture group was treated with exercise training combined with scalp acupuncture.The treatment time was 4 weeks.Morris water maze test was performed after treatment,positioning navigation test was carried out in the first five days,and space exploration test was carried out on the sixth day after the escape platform of rats was removed.Morris water maze test was used to observe the effects of various treatments on cognitive learning ability of VaD rats.At the end of treatment,some rats were decapitated and the hippocampal tissue was removed.The expressions of pro-inflammatory factors IL-6,IL-17,IL-1 β,TNF-α and anti-inflammatory factors IL-10,TGF-β genes mRNA in the hippocampus of each group were detected by RT-PCR method.The brains of some rats were taken after perfusion,and the histomorphological changes of hippocampal CA1 region were observed by HE staining.Results:(1)Localization navigation test: compared with the sham operation group,the escape latency of the model group and each treatment group increased before treatment(P < 0.05),and after treatment,compared with the model group,the escape latency of each treatment group decreased(P < 0.05).Among them,the scalp acupuncture exercise acupuncture group was the most significant,and the difference was statistically significant(P < 0.05).(2)Space exploration test: compared with the sham operation group,the time of reaching the escape platform for the first time in the model group and each treatment group was significantly increased before treatment(P < 0.05),while the times of crossing the platform and the stay time of the platform were significantly decreased(P < 0.05).Compared with the model group,the time of reaching the escape platform for the first time in each treatment group was significantly lower than that in the model group(P < 0.05),while the times of passing through the platform and staying time on the platform were significantly increased(P < 0.05),especially in the scalp acupuncture exercise acupuncture group.The difference was statistically significant(P < 0.05).(3)RT-PCR detection of related inflammatory factors: compared with the sham operation group,the relative expression of IL-6,IL-17,IL-1 β and TNF-α gene mRNA in the hippocampus of the model group was significantly increased,and after treatment,compared with the model group,the relative expression of related proteins decreased,especially in the scalp acupuncture group,and the difference was statistically significant.(4)RT-PCR detection of related anti-inflammatory factors: compared with the sham operation group,the relative expression of anti-inflammatory factors IL-10 and TGF-β gene mRNA in the hippocampus of the model group had no significant change,but after treatment,the relative expression of related proteins in each treatment group increased compared with the model group,especially in the scalp acupuncture group,and the difference was statistically significant.(5)Correlation analysis between hippocampal inflammatory factors and escape latency: the relative expression of related inflammatory factor gene mRNA was positively correlated with escape latency,and the relative expression of related anti-inflammatory factor gene mRNA was negatively correlated with escape latency.(6)HE staining: in the sham operation group,the tissue structure of hippocampal CA1 region was complete and clear,the cells arranged regularly,the shape was normal,the outline was clear,and the nucleolus could be clearly identified;in the model group,the arrangement of tissue cells in hippocampal CA1 region was vague and confused,the number of normal cells decreased,necrotic neurons and obvious edema could be seen,and some nucleolus pyknosis and deep staining could not be seen.Compared with the model group,the CA1 area of brain in the exercise training group,scalp acupuncture group and scalp acupuncture exercise acupuncture group was relatively regular,the outline was relatively clear,the number of normal cells increased,the interstitial edema was alleviated,the necrotic neurons decreased and the nucleolus was clearer than that in the model group.Among them,the scalp acupuncture exercise acupuncture group has the best structure.Conclusions:1.Both exercise training and scalp acupuncture can significantly improve the learning and memory ability and spatial exploration ability of VaD model rats,and can also improve the pathological morphology of hippocampal tissue of VaD model rats,but the therapeutic effect of the combination of the two is more significant.2.Abnormal secretion of inflammatory factors in hippocampus may be one of the pathogenesis of VaD.The mRNA levels of inflammatory factors such as IL-6,IL-17,IL-1 β and TNF-α in the hippocampus of VaD rats were significantly increased.After scalp acupuncture treatment,the levels of inflammatory factors IL-6,IL-17,IL-1 β and TNF-α decreased,while the levels of anti-inflammatory factors IL-10 and TGF-β increased.The cognitive learning ability of VaD rats was also greatly improved.It is suggested that scalp acupuncture can regulate the level of inflammatory factors in hippocampus and improve the cognitive ability of VaD rats. |
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