补肾活血颗粒干预帕金森病模型大鼠黑质VEGF及纹状体Trkb、Tau表达的影响

目的:探讨补肾活血颗粒对帕金森病模型大鼠黑质VEGF及纹状体Trkb、Tau表达的影响。方法:33只雄性SD大鼠随机分为生理盐水正常组、6-羟基多巴胺(6-OHDA)模型组和补肾活血颗粒治疗组,应用免疫组织化学染色技术观察大鼠黑质细胞VEGF及纹状体Trkb、Tau表达的变化情况。结果:正常组与模型组VEGF比较,两者表达相近,且P>0.05,差异无统计学意义;治疗组与模型组比较,治疗组VEGF表达显著增强,且P<0.01,差异有统计学意义;治疗组与正常组比较,治疗组VEGF表达明显增强且P<0.01,差异有统计学意义;正常组与模型组Trkb比较,模型组Trkb表达增强,P<0.01差异有统计学意义;治疗组与模型组比较,治疗组Trkb表达显著增强,且P<0.01,差异有统计学意义;治疗组与正常组比较,治疗组Trkb表达明显增强且P<0.01,差异有统计学意义;正常组与治疗组Tau比较,正常组表达增强且P<0.01,差异有统计学意义;正常组与模型组比较,模型组Tau表达显著增强,且P<0.01,差异有统计学意义;模型组与治疗组比较,治疗组Tau表达明显降低且P<0.01,差异有统计学意义。结论:补肾活血颗粒治疗帕金森病的作用机制之一可能是通过上调与神经保护因子相关的VEGF、Trkb蛋白表达,营养神经细胞增强血管新生和下调与蛋白质凝聚相关的Tau蛋白表达抑制细胞凋亡保护神经元有关。

补肾活血颗粒对帕金森病模型大鼠黑质纹状体bcl-2、bax表达的影响

目的探讨补肾活血颗粒对帕金森病模型大鼠黑质纹状体中B淋巴细胞瘤-2蛋白(B-cell lymphoma-2,bcl-2)、bax表达的影响。方法 33只雄性SD大鼠随机分为生理盐水正常组、6-羟基多巴胺模型组和补肾活血颗粒治疗组,应用免疫组织化学染色技术观察大鼠黑质纹状体中bcl-2、bax表达的变化情况。结果模型组bcl-2在黑质和纹状体的细胞阳性表达率增殖指数(proliferation index,PI)分别为0.43±0.022、1.14±0.057,均低于正常组(均P<0.01)和治疗组(均P<0.01)。模型组bax在黑质和纹状体的PI值分别为2.07±0.104、2.86±0.143,均高于正常组(均P<0.01)和治疗组(均P<0.01)。结论补肾活血颗粒治疗帕金森病的疗效机制可能与上调帕金森病大鼠黑质纹状体系统bcl-2蛋白表达及下调bax蛋白表达有关。

铁对大鼠黑质神经递质影响的初步研究

目的:探讨Fe3+/Fe2+对大鼠黑质神经递质影响。方法:黑质内分别注射:FeCl3、FeCl2和6-羟多巴胺(6-OHDA),四周后应用阿朴吗啡诱发旋转行为、高压液相色谱分析检测黑质内多巴胺、去甲肾上腺素、肾上腺素和5-羟色胺水平。结果:注射FeCl3的大鼠出现向损毁同侧的旋转行为;注射FeCl3和FeCl2侧黑质内多巴胺水平显著降低,去甲肾上腺素、肾上腺素和5-羟色胺水平不受影响。结论:Fe3+/Fe2+对大鼠黑质多巴胺能神经元具有特异性的毒性作用。

补肾活血颗粒对帕金森病模型大鼠黑质细胞TrkB及Tau表达的影响

目的:探讨补肾活血颗粒对帕金森病模型大鼠黑质中TrkB及Tau表达的影响。方法:33只雄性SD大鼠随机分为正常组、模型组和补肾活血颗粒治疗组,应用免疫组织化学染色技术观察大鼠黑质中TrkB、Tau表达的变化情况。结果:与正常组及模型组比较,治疗组TrkB表达显著增强(P<0.01);与正常组比较,模型组Tau表达显著增强(P<0.01);与模型组比较,治疗组Tau表达明显降低(P<0.01)。结论:补肾活血颗粒治疗帕金森病的疗效机制可能与上调模型大鼠黑质细胞中的TrkB表达,强化了神经修复损伤因子作用及下调Tau蛋白,减轻细胞内蛋白质凝聚表达有关。

大鼠脑黑质直接注射Aβ引起多巴胺神经元丢失

β-淀粉样蛋白(Aβ)沉积是AD病人脑内老年斑周边神经元变性和死亡的主要原因,帕金森病伴痴呆症也与Aβ的沉积相关。该研究直接将Aβ注射到大鼠脑黑质区,通过免疫染色与脑黑质TH-Positive的神经元统计计数,来检测Aβ对多巴胺神经元损伤及小胶质细胞的激活情况。

Copper (Cu^(2+)) induces degeneration of dopaminergic neurons in the nigrostriatal system of rats

Objective To study the effects of intranigral injection of different doses of CuSO45H2O on dopaminergic neuron in the nigrostriatal system of rats. Methods Wistar rats were divided into four groups, including control group, 10 nmol, 50 nmol and 200 nmol copper injected into left substantia nigra （SN） groups. Seven days after the intranigral injection of copper, dopamine （DA） contents in the striatum （Str） were measured by high performance lipid chromotophotography （HPLC）; the density of tyrosine hydroxylase （TH） positive axons in the Str was measured by TH staining method; TH and Caspase-3 mRNA expression in the SN were measured by semi-quantitative RT-PCR. We detected the activity of superoxide dismutase （SOD） in the lesioned midbrain of rats using biochemical methods. Results DA and its metabolites contents had no significant difference between control group and low dose （10 nmol） copper group. But from 50 nmol copper group, DA contents in the lesioned sides were reduced with the increase in the copper doses injected, showing a significant linear correlation （F = 34.16, P 〈 0.01）. In the 50 nmol copper group, TH positive axons in the Str decreased compared with those of the control and unlesioned sides （F = 121.9, P 〈 0.01）. In the 50 nmol copper group, TH mRNA expression decreased （t =3.12, P 〈 0.01） while Caspase-3 mRNA expression increased （t =8.96, P 〈 0.01） in the SN compared with the control. SOD activity decreased in the midbmin of rots treated with 50 nmol copper compared with that of the control （t = 2.33, P〈0.01）. Conclusion Copper could induce damage of dopaminergic neurons in the SN of rats through destroying antioxidant defenses and promoting apoptosis.

Experimental study on the effect of An-pressing and Rou-kneading Huantiao (GB 30) on certain brain nuclei of pleasure circuits in rats with chronic neuralgia

Objective:To explore the central neurobiological mechanisms of pleasure effect on rats with neuralgia treated by tuina manipulations of An-pressing and Rou-kneading Huantiao (GB 30).Methods:A total of 64 male Sprague-Dawley (SD) rats were used in this study.Eighteen rats were randomly selected as a normal group,and the other 46 rats were used to duplicate the chronic constriction injury (CCI) model.Ten rats failed in modeling and 36 rats succeeded.These 36 rats were then randomly divided into a model group and a tuina group,with 18 rats in each group.The rats in the normal group and the model group did not receive any interventions,while those in the tuina group received An-pressing and Rou-kneading Huantiao (GB 30),1 min for each time,once a day,3 weeks in total.Heating tests were evaluated to observe the change of pain-sensitivity score before intervention,1 week after intervention,2 weeks after intervention,and 3 weeks after intervention.After 1 week of intervention,2 weeks of intervention,and 3 weeks of intervention,6 rats were randomly selected from each group respectively for brain extraction.The change of Nissl's body and β-endorphin in the accumbens nucleus as well as amygdaloid nucleus of pleasure circuits,and pro-opiomelanocortin (POMC) in the arcuate nucleus were analyzed by methods of histochemistry and molecular biology.Results:After modeling,the pain-sensitivity scores of the tuina group and the model group were statistically different from the score of the normal group (both P<0.05).After An-pressing and Rou-kneading Huantiao (GB 30) for one week,the pain-sensitivity score of the tuina group had statistical difference compared with that of the model group (P<0.05).At each different time point:the amounts of Nissl's body in accumbens nucleus and amygdaloid nucleus of the tuina group were significantly more than those of the model group (all P<0.01).Besides,the numbers of β-endorphin immunoreactive cells in the accumbens nucleus and amygdaloid nucleus of the rats in the tuina group were significantly higher than those in the model group (all P<0.01),and so was the expression of POMC in arcuate nucleus (all P<0.01).Conclusion:An-pressing and Rou-kneading Huantiao (GB 30),where the sciatic nerve is ligated,can reduce pain-sensitivity score and increase pain tolerance value of rats with chronic neuralgia.It can increase the activity of neurons in accumbens nucleus and amygdaloid nucleus of pleasure circuits,which indicates that the analgesia effect of tuina therapy may correlate with pleasure effect,and also reveals a part of neurobiological mechanisms of neuralgia.