朱琏兴奋Ⅰ型针法对糖尿病膀胱模型大鼠脑源性神经营养因子及其受体TrkB和环磷酸腺苷表达的影响

Effects of Zhu Lian stimulant Type I acupuncture on the expression of brain-derived neurotrophic factor and its receptor TrkB and tissue homogenate cyclic adenosine phosphate in rats with diabetic bladder

目的观察朱琏兴奋Ⅰ型针法对糖尿病膀胱(DCP)模型大鼠脑源性神经营养因子(BDNF)及其受体酪氨酸激酶受体B(TrkB)、环磷酸腺苷(cAMP)表达的影响,探讨朱琏兴奋Ⅰ型针法治疗DCP的作用机制。方法将50只SD大鼠按随机数字表法分为对照组、模型组、西药组、普通针刺组、针法治疗组,每组10只。除对照组外,其余各组腹腔注射链脲佐菌素(STZ)制备DCP大鼠模型。西药组灌胃甲钴胺;选取大鼠"中极""三阴交""列缺""太冲"穴,普通针刺组行普通针刺手法,针法治疗组行朱琏兴奋Ⅰ型针法干预,1次/d,30 min/次。干预4周后取材。测定各组大鼠膀胱最大容量、残余尿量、膀胱湿重;HE染色观察各组大鼠膀胱组织形态;免疫组化染色检测BDNF表达;Western blot法检测cAMP蛋白表达;ELISA法测定TrkB水平;实时荧光定量PCR检测BDNF mRNA、cAMP mRNA水平。结果与模型组比较,西药组、普通针刺组、针法治疗组大鼠膀胱最大容量、残余尿量、膀胱湿重降低(P<0.01),且西药组、针法治疗组低于普通针刺组(P<0.01);西药组、普通针刺组、针法治疗组BDNF mRNA及蛋白、cAMP mRNA及蛋白表达升高(P<0.05),TrkB水平升高,且西药组、针法治疗组高于普通针刺组(P<0.05)。结论朱琏兴奋Ⅰ型针法对糖尿病大鼠膀胱功能具有保护作用,其机制可能与上调BDNF蛋白及mRNA、TrkB、cAMP蛋白及mRNA水平有关。

Objective To observe the effects of Zhulian stimulant type I acupuncture on the expression of brain-derived neurotrophic factor(BDNF)and its receptor TrkB and tissue homogenate cyclic adenosine phosphate(cAMP)in rats with diabetic bladder(DCP);To explore the mechanism of Zhulian stimulant type I acupuncture on DCP.Methods Totally 5o SD rats were divided into control group,model group,Western medicine group,ordinary acupuncture group,Zhulian stimulant type I acupuncture treatment group(acupuncture treatment group)according to random number table method,with 10 rats in each group.DCP rat model was prepared by intraperitoneal injection of streptozotocin(STZ),except for the control group.The Western medicine group was given mecobalamine for gavage;acupoints of"Zhongji","Sanyinjiao","Liechou"and"Taichong"were selected.The ordinary acupuncture group was treated with ordinary acupuncture technique,and the acupuncture treatment group was treated with Zhulian stimulant type I acupuncture,1 time/d,30 minutes/time.Samples were taken after 4 weeks of treatment.The maximum bladder volume,residual urine volume and wet weight of the bladder were detected.The morphology of rat bladder was observed by HE staining.The expression level of BDNF was detected by immunohistochemistry.The expression of cAMP was detected by Western blot.The level of TrkB was determined by ELISA.Real-time fluorescence quantitative PCR(RT-PCR)was used to detect the mRNA expressions of BDNF and cAMP.Results Compared with model group,maximum bladder volume,residual urine volume and wet weight of bladder in Western medicine group,ordinary acupuncture group and acupuncture treatment group decreased(P<0.01),and those in Western medicine group and acupuncture treatment group were lower than those in ordinary acupuncture group(P<0.01).The expressions of BDNF mRNA and protein,cAMP mRNA and protein in Western medicine group,ordinary acupuncture group and acupuncture treatment group increased(P<0.05),and the level of TrkB increased,and the Western medicine group and acupuncture treatment group were higher than that in ordinary acupuncture group(P<0.05).Conclusions Zhuliping stimulant type I acupuncture has a protective effect on the bladder function of diabetic rats.The mechanism may be related to the up-regulation of BDNF and mRNA,TrkB,cAMP and mRNAexpressions.