穴位埋线对慢性应激复合衰老模型大鼠海马自噬的影响  

Influence of Acupoint Catgut Embedding on Autophagy of Hippocampal Neuron in Aging Rat Model Induced by Multi-Chronic Stress

作　　者：袁玉梅 范春[2] 林琨伟 陈海桥 吴剑萍 谢学铭[2] 张宏[2] YUAN Yumei;FAN Chun;LIN Kunwei;CHEN Haiqiao;WU Jianping;XIE Xueming;ZHANG Zhong(Baoan Hospital of TCM in Shenzhen, Shenzhen 518133, Guangdong, China;Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong, China)

机构地区：[1]深圳市宝安中医院(集团),广东深圳518133 [2]广州中医药大学,广东广州510405

出　　处：《辽宁中医杂志》2018年第2期401-404,I0004,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：广东省高水平大学建设项目(AFD018161Z0204);国家自然科学基金项目(81573912);广东省自然科学基金项目(2015A030313356);深圳市福田区公益科技项目(FTW2015003)

摘　　要：目的：通过观察穴位埋线对慢性应激复合衰老大鼠神经元组织形态及海马自噬影响,探讨慢性应激的促衰老作用及穴位埋线的抗衰机制。方法：将3月龄雄性SD大鼠随机分为空白对照组、模型一组、模型二组、穴位埋线组、假埋线组,每组9只。模型一组予D-半乳糖[120 mg/（kg·d）]腹腔注射;模型二组、穴位埋线组、假埋线组予慢性应激复合D-半乳糖[120 mg/（kg·d）]腹腔注射;穴位埋线组每周埋线一次,假埋线组每周假埋线一次。实验结束后对海马进行HE及尼氏染色,观察海马的组织形态结构;q PCR检测海马ULK1、Beclin1、LC3-II、P62 mRNA的表达。结果：（1）与空白对照组相比,模型一组大鼠海马神经元组织结构疏松,排列紊乱,细胞层次减少,部分神经元萎缩,尼氏体减少,模型二组的大鼠海马神经元组织更加疏松,细胞间隙显著增大,大量神经元萎缩深染,神经元数量进一步减少,尼氏体减少甚至消失。穴位埋线组大鼠海马神经元排列较整齐致密,细胞间隙稍增大,尼氏体清晰;而假埋线组海马神经元结构较模型二组大鼠并无明显差异。（2）与空白对照组相比,模型一组大鼠海马神经元ULK1、Beclin1 mRNA的表达有升高趋势,LC3-II、P62 mRNA表达升高;与模型一组相比,模型二组的大鼠海马神经元LC3-II、P62 mRNA表达进一步升高,ULK1、Beclin1 mRNA表达有升高趋势;与模型二组相比,穴位埋线组大鼠海马神经元ULK1、Beclin1、LC3-II、P62 mRNA表达均降低,假埋线组大鼠海马神经元ULK1、Beclin1、LC3-II、P62 mRNA表达则无明显差异。结论：慢性应激确实会通过影响衰老大鼠海马神经元的自噬功能从而加速海马的老化,穴位埋线可以拮抗应激性衰老机体海马神经元的自噬流损伤,从而保护海马神经元,延缓衰老。Objective： To observe the influence of acupoint catgut embedding on histomorphology and autophagy of hippocampal neuron in aging rat model induced by multi-chronic stress. Methods： Forty-five healthy SPF SD rats aged 3 months were randomly divided into blank control group,model group 1,model group 2,acupoint catgut embedding group and shamacupoint catgut embedding group. The model group 1 was given intraperitoneal injection with D-galactos[120 mg/（kg/d） ]while model group 2with acupoint catgut embedding group and sham acupoint catgut embedding group were given intraperitoneal injection with D-galactos[120 mg/（kg/d） ] and multi-chronic stress. Otherwise,the acupoint catgut embedding group was also given acupoint catgut embedding once a day for 6 weeks while sham group was given sham acupoint catgut embedding once a day for 6 weeks. At the end of the experiment,the hippocampus was cut into sections to be stained with H. E. and nissl technique. Also,the rats were executed for the examination of ULK1,Beclin1,LC3-II and P62 mRNA expressions of hippocampus. Results（1） Compared with the control group,hippocampal structure of model group 1 was loose and disordered,cellular level decreased,part of the neuronatrophied and austenite reduced. Hippocampal structure of model group 2 was more loose and disordered. A large number of neuronal atrophied,the number of neurons decreased further,austenite reduced or even disappear. In the acupoint catgut embedding group,hippocampal neurons were dense,intercellular space increased slightly,austenite was clear. Hippocampal structure of sham acupoint catgut embedding group was like that of model group 2.（2） Compared with the control group,the expressions of ULK1 and Beclin1 mRNA in model group 1 presented a rising trend and the expressions of LC3-II and P62 mRNA increased. Compared with the model group 1,the expressions of LC3-II and P62 mRNA in model group 2 increased futher. The expressions of ULK1 and Beclin1 mRNA presented a rising trend. Compared with the model

关 键 词：衰老 应激 海马 自噬 穴位埋线 

分 类 号：R244.8[医药卫生—针灸推拿学]