穴位埋线对失眠大鼠认知功能及5-HT、DA、GABA水平的影响

Effects of acupoint catgut embedding on cognitive function and levels of 5-HT,DA and GABA in insomnia rats

目的观察穴位埋线治疗对氯苯丙氨酸(PCPA)失眠模型大鼠下丘脑5-羟色胺(5-HT)、多巴胺(DA)、γ-氨基丁酸(GABA)表达的影响,探讨穴位埋线治疗失眠的可能机制。方法选取24只SPF级SD雄性大鼠,6只作为空白组,其余18只采用腹腔注射PCPA混悬液制作失眠模型。造模成功后,将模型大鼠随机分为模型对照组、针刺组和穴位埋线组,各6只。空白组和模型对照组大鼠仅抓取,不予干预;针刺组针刺百会、神门(双侧)和三阴交(双侧),1 d/次,连续14 d;埋线组进行埋线,选穴同针刺组,7 d/次,共2次。采用戊巴比妥钠协同睡眠实验观察大鼠睡眠潜伏期和睡眠持续时间;Morris水迷宫实验评价大鼠空间学习记忆能力;苏木精-伊红染色(HE)观察下丘脑组织神经元细胞形态;酶联免疫吸附试验(ELISA)检测下丘脑组织5-HT、DA、GABA蛋白表达。结果与空白组比较,模型对照组大鼠下丘脑细胞呈现空泡样变化、大量炎性细胞浸润,睡眠潜伏期延长,睡眠持续时间缩短,逃逸潜伏期增加、穿越隐藏平台次数减少,下丘脑组织GABA、5-HT蛋白表达降低、DA蛋白表达升高(P<0.05);与模型对照组比较,针刺组、穴位埋线组大鼠下丘脑正常神经元细胞数量增加、空包化状等神经炎症均有不同程度改善,睡眠潜伏期缩短、睡眠持续时间延长,逃逸潜伏期减少、穿越隐藏平台次数增加,下丘脑组织GABA、5-HT蛋白表达升高、DA蛋白表达降低(P<0.05);与针刺组比较,穴位埋线组大鼠下丘脑组织DA蛋白表达下降(P<0.05),睡眠潜伏期、睡眠持续时间、逃逸潜伏期、穿越隐藏平台次数、GABA、5-HT蛋白表达差异均无统计学意义(P>0.05)。结论穴位埋线治疗可有效缩短PCPA失眠模型大鼠睡眠潜伏期、延长睡眠持续时间,提高空间学习记忆能力等认知功能,其机制可能与改善下丘脑组织病理形态、调节下丘脑内神经递质GABA、5-HT及DA水平有关。

Objective To observe effects of catgut implantation at acupoint treatment on expression of 5-hydroxytryptamine(5-HT),dopamine(DA)andγ-aminobutyric acid(GABA)in hypothalamus of PCPA insomnia model rats,and to investigate possibilities of mechanisms of catgut implantation in treatment of insomnia.Methods Twenty-four SPF SD male rats were selected,6 of them served as blank group,and the other 18 rats were made insomnia model by intraperitoneal injection of PCPA suspension.After successful modeling,the model rats were randomly divided into model control group,acupuncture group and acupoint catgut embedding group,6 rats each.The rats in control group and model group only grasped without intervention;acupuncture group needled Baihui,Shenmen(bilaterally)and Sanyinjiao(bilaterally),1 d/times for 14 days;burying thread group buried threads,with the same points as acupuncture group,for 7 d/times twice in total.Sodium pentobarbital synergistic sleep experiment observed sleep latency and sleep duration in rats;the Morris water maze experiment investigated the rats′capacity of spatial cognition;HE staining analyzed the neuronal cell of hypothalamus tissues;5-HT,DA,and GABA protein contents of hypothalamus tissues tested with ELISA method.Results Compared with model group,the sleep latency of rats in each treatment group was shortened and the sleep duration was prolonged(P<0.05).Compared with model group,the escape latency of rats in each treatment group were decreased and the number of times of crossing the hidden platform wereincreased(P<0.05).Compared with model group,the nerve cells in the hypothalamus area were arranged in a messy and loose manner,the gap was increased,and the neuroinflammation such as empty package was improved after the treatment.Compared with model group,the concentration of GABA and 5-HT protein in each treatment group was increased,the concentration of DA protein was decreased(P<0.05),and the concentration of DA protein was decreased significantly(P<0.05).Conclusion Catgut embedding therapy can effectively improve insomnia,anxiety symptoms and cognitive function such as learning and memory in insomnia rats.The mechanism may be related to the regulation of neurotransmitters GABA,5-HT and DA concentrations in the hypothalamus.