“双固一通”电针法对老年阳虚模型大鼠海马超微结构及BDNF含量和PKCmRNA表达的影响

Effects of Double-Reinforcing and One-Unblocking of Acupuncture Method on Hippocampus Synaptic Structure and Content of BDNF and Expression of PKC mRNA in Aging Rats with Yang Deficiency

目的观察"双固一通"电针法对老年阳虚模型海马CA1区突触形态结构海马组织内神经细胞营养因子(BDNF)含量和PKCmRNA表达的影响。方法选用SD雄性大鼠随机分为正常组、模型组、电针组和对照组。模型组、电针组和对照组大鼠每日颈背部皮下注射125mg/kg D-半乳糖,连续40d,然后再进行后腿肌肉注射氢化可的松1.5mg/100g,连续7d,成功诱导出衰老模型大鼠。正常组大鼠每日皮下注射等量0.9%生理盐水1次,连续47d。电针组采用"双固一通"电针法持续治疗4周,对照组分别取"中极"、"阴陵泉"、"印堂"作为对照腧穴,治疗时间与电针组相同,正常组和模型组不进行治疗。采用电镜结合图像分析对大鼠海马CA1区突触形态结构进行观察;运用放射免疫法检测大鼠海马组织BDNF含量;RT-PCR检测PKCmRNA含量。结果与正常组比较,模型组大鼠海马突触活性区长度变短,突触后致密物厚度变薄,突触间隙宽度变大,组间比较具有显著差异(P<0.01),海马BDNF含量减少(P<0.05),海马中PKCmRNA的表达水平明显降低(P<0.01);与模型组比较,电针组大鼠的突触后致密物厚度增加,突触间隙宽度减小,差异显著(P<0.01),海马BDNF含量明显提高(P<0.05)、PKCmRNA的表达水平明显升高(P<0.01);与电针组比较,对照组大鼠海马突触活性区长度缩短(P<0.01),突触界面曲率降低(P<0.05),海马组织内BDNF含量降低(P<0.05)、PKCmRNA的表达降低(P<0.05)。结论 "双固一通"电针法可改善老年阳虚模型大鼠海马突触活性区长度,突触后致密物厚度和突触间隙宽度,并且能够调节和提高大鼠海马中PKCmRNA的表达水平及BDNF含量。

Objective To observe the effects of Double-reinforcing and One-unblocking of acupuncture on hippocampus synaptic structure,the content of BDNF,and the expression of PKCmRNA of aging model rats with Yang-deficiency.Methods 48 SD rats were randomly divided into four groups：Normal Group,Model Group,Double-reinforcing and One-uublocking of electro-acupuncture Group （EA）,and electro-acupuncture control Group （EC）.Every rats in Model Group,EA Group,and EC Group were injected with D-galactose subcutaneously for 40 days,to induce the sub-acute aging model rats.Then except the rats in Normal Group,all rats were injected with hydrocortisone for 7davs,to induce the Yang-deficiencv symptom on the aging model rats.The rats in Normal Group were injected of same amount of saline.Rats in EA Group were treated with Double-reinforcing and One-unblocking of electro-acupuncture,and EC Group were stimulated with ＂ Yintang＂ ＂Zhongji YinLingquan＂,each treatment were last for 4 weeks.Synaptic structural parameters were analysised by electron microscopy and biological image analysis system; detect the content of BDNF by radioimmunity method; RT-PCR examines the PKC mRNA expression.Results In comparison with the Normal Group,the length of active zones and the thickness of PSD are all decreased,the synaptic cleft width enlarged significantly in the Model group（P 〈0.01）,the content of BDNF declined remarkably in Model Group（P 〈 0.05）,the expression of PKCmRNA declined remarkably in Model Group（P 〈 0.01）; In comparison with the Model Group,the thickness of PSD increased,and the synaptic cleft width decreased remarkably in the EA Group （P 〈 0.01）,the content of BDNF in EA Group increased（P 〈 0.05）,the expression of PKCmRNA raised remarkably in EA Group （P 〈 0.01） ; In comparison with the EA Group,the length of active zones of PSD decreased in EC Group（P 〈0.05）,the content of BDNF in EC Groupdecreased（P 〈0.05）,the content of PKCmRNA in ECGroup decreased （P 〈 0.05）.Conclusion The above results suggest that the effects of Double-reinforcing and One-unblocking of acupuncture method can improve the hippocampal synaptic structural parameters; and the treatment can adjust and regulate content of BDNF and the PKC mRNA expression in aging model rats with Yang-deficiency.