头穴丛刺联合取食训练对2型糖尿病认知障碍大鼠海马神经元损伤的治疗作用

Therapeutic effect of scalp clustering acupuncture combined with feeding training on hippocampal neuron damage in T2DM rats with cognitive impairment

目的观察头穴丛刺联合取食训练对2型糖尿病(T2DM)认知障碍大鼠海马神经元损伤的治疗作用。方法健康雄性SD大鼠72只,随机分为对照组、T2DM组、模型组、头穴丛刺组、取食训练组、联合组,每组12只。对照组常规饲养,T2DM组制备T2DM模型,模型组、头穴丛刺组、取食训练组和联合组制作T2DM认知障碍模型。头穴丛刺组予头穴丛刺,取食训练组进行取食训练,联合组在头穴丛刺长留针期间进行取食训练;对照组、T2DM组、模型组不予任何干预。分别于造模14、28 d后,采用Morris水迷宫评价大鼠学习和记忆能力;处死大鼠取海马组织,透射电镜下观察海马组织超微结构,采用ELISA法检测超氧化物歧化酶(SOD)和丙二醛(MDA),采用Western blotting法检测脑源性神经营养因子(BDNF)蛋白。结果与T2DM组相比,模型组逃避潜伏期延长,穿越平台次数和目标象限停留时间缩短,海马神经元髓鞘不完整,线粒体呈空泡化倾向、嵴溶解消失,海马组织SOD活性、BDNF蛋白表达降低,MDA含量增加(P均<0.05)。与模型组相比,头穴丛刺组、取食训练组、联合组逃避潜伏期缩短,穿越平台次数和目标象限停留时间延长,海马神经元超微结构改变减轻,海马组织SOD活性、BDNF蛋白表达增加,MDA含量减少(P均<0.05)。联合组各项优于头穴丛刺组和取食训练组,且造模28 d指标优于造模14 d(P均<0.05)。结论头穴丛刺联合取食训练治疗能够减轻氧化应激损伤,修复海马神经元超微结构,改善T2DM认知障碍大鼠学习记忆功能,且治疗时间越长,效果越显著。

Objective To observe the therapeutic effect of scalp clustering acupuncture combined with feeding training on hippocampal neuron damage in type 2 diabetes mellitus(T2DM)rats with cognitive impairment.Methods Seventy-two healthy male SD rats were randomly divided into six groups,including the normal control group,the T2DM group,the model group,the scalp clustering acupuncture group,the feeding training group,and the scalp clustering acupuncture group combined with feeding training group,with 12 rats in each group.Rats in the normal control group were routinely fed;in the T2DM group,we prepared the T2DM models;in the other four groups,we prepared the cognitive impairment models of T2DM.Rats in the normal control group,the T2DM group and the model group were not intervened.Rats in the scalp clustering acupuncture group were given scalp clustering acupuncture intervention,and rats in the feeding training group were given feeding training intervention,and rats in the scalp clustering acupuncture+feeding training group were given feeding training intervention along with scalp clustering acupuncture.After 14 and 28 days of intervention,the Morris water maze(MWM)test was used to observe the learning and memory ability of the model rats,ELISA was used to detect the content of superoxide orgotein dismutase(SOD)and maleic dialdehyde(MDA),and Western blotting was used to detect the protein expression of brain-derived neurotrophic factor(BDNF).Results Compared with the T2DM group,the escape latency was prolonged,and the number of times crossing platforms was smaller and the residence time in the target quadrant was significantly shorter,the myelin sheath of hippocampal neurons was incomplete,mitochondria tended to be vacuolated,cristae dissolved and disappeared,the activity of SOD and the expression of BDNF protein in hippocampal tissues decreased,and the content of MDA increased in the model group(all P<0.05).The scalp clustering acupuncture,feeding training group,and scalp clustering acupuncture combined with feeding training group had significantly shorter escape latency,significantly larger number of times crossing platforms and longer residence time in the target quadrant,alleviated ultrastructural changes of hippocampal neurons,and significantly increased hippocampal tissue SOD activity,BDNF protein expression level,and significantly decreased MDA content as compared with the model group(all P<0.05).The indexes in the scalp clustering acupuncture+feeding training group were superior to those of the scalp clustering acupuncture group or feeding training group alone,and the indexes at 28 days of intervention were superior to those at 14 days of intervention(all P<0.05).Conclusion Scalp clustering acupuncture combined with feeding training improves the learning memory function of T2DM cognitively impaired rats by attenuating peroxidative kinetic damage of cells and repairing the ultrastructure of hippocampal neurons,and the longer the treatment time,the more significant the effect is.