| Parkinson’s disease (PD) is a chronic progressive neurodegenerative in central nervous system characterized by an apoptosis and selective loss of nigrostriatal dopaminergic neurous. PD is caused by environmental toxin and genetic factor and so on. Clinical manifestations of this complex disease include static trembling, myotomia, bradukinesia, posture instability and so on. Because of the complex pathology and pathogenesis, there are no ideal therapies of PD at present. The current medications are efficient in the early stage of the disease but it will bring an unbearable side-effect such as movement disorder after a long-term use. Growing attention has been paid on traditional medicine such as acupuncture treatment because of its favorable curative effect and low adverse reactions. According to the classical theory in traditional Chinese medicine, brain is the house of mental activity and the marrow sea, so scalp acupuncture has been widely used in clinic treatment of PD and chorea trembling control area is the special treatment area. Studies indicate that Brain derived neurotrophic factor (BDNF) is a protein to provide nutrients and prevent dopaminergic neurons from losing and damaging, and BDNF has been a hotpot in researches of PD.To explore the effect and possible mechanism of needling chorea trembling control area in treating PD and provide scientific foundation to clinical treatment, we observed the changes of dopaminergic neurons and the expression of BDNF and TrkB pathway in MPTP-induced PD mice.This study mainly divides into two parts:(1) To investigate the neuroprotect effect on needling chorea trembling control area on dopaminergic neurons, we observed ethology and changes in dopaminergic neurons in PD mice.(2) To explore the curative mechanism of acupuncture in PD mice, we observed the influence of needling chorea trembling control area on the activity of BDNF and TrkB receptor as well as molecules in signaling pathways downstream.We use C57BL/6J mice in the experiment. After a training of climbing rob method, we selected mice limbs and climb down from the pole smoothly and divided them into five groups randomly:normal control group; model control group; medopar group; needling group; needling combine with medopar group. In the experiment, all the mice except normal control group were induced to PD model by intraperitoneal injection of0.2%MPTP,20mg/(kg· d) per day and continuous injection for5days, then treated with corresponding treatment. The animals were killed after28days from they first injected MPTP.The main experiment results are as follows:1. Effects of needling chorea trembling control area on behavior of PD mice.Mice showed some acute effects such as movement disorder, piloerection and tail-erecting after MPTP injection for5to30minutes. There are symptoms of weight loss and color dim after5times injections. We scored the time of climbling rob and the result demonstrate that compared with nomal group, the score model group is obviously lower (P<0.05); then needling, medopar, and needling combined with medopar group could improve behavior of PD mice obviously (P<0.05).2. Effects of needling chorea trembling control area on pathology of substantia nigra in PD mice.The result shows that the number of neurons in the substantia nigra decreased and many atrophy, deep dyeing neurons appeared in the model group compared with normal group. The number of neurons and normal form neurons in substantia nigra increased in three treatment group compared "with model group.3. Effects of needling chorea trembling control area on the number of TH positive neurons of PD mice.SP immunohistochemistry was used in this experiment and the number of TH (Tyrosine hydroxylase) positive neurons was counted each group. Compared with normal group, the number of TH positive neurons in other four groups decreased distinctly (P<0.05). The number of TH positive neurons in needling combined with medopar group is evidently increased compared with medopar group (P<0.05).4. Effects of needling chorea trembling control area on the expression of BDNF of PD mice.The result shows that expression of BDNF was obviously decreased in model group compared with normal group (P<0.05); needling combined with medopar group was significantly higher than the model group (P<0.05).5. Effects of needling chorea trembling control area on the number of TrkB receptor of PD mice.The result shows that in the substantial nigra, compared with normal group, the expression in the model and medopar group decreased significantly (P<0.05); compared with model group, needling group and needling combined with medopar group increased (P<0.05); compared with medopar group, needling group increased obviously (P<0.05); compared with needling group, needling combined with medopar group decreased (P<0.05).In the CA3hippocampus area, the expression of TrkB receptor in model group was obviously less than in the normal group (P<0.05); expression in needling group of CA1-CA4area increased compared with normal and model group (P<0.05), medopar group decreased compared with needling group (P<0.05); in CA1and CA2area, the expression in needling combined with medopar group increased compared with model group (P<0.05).6. Effects of needling chorea trembling control area on the expression of PSD-95of PD mice.The result shows that compared with normal group, the expression in model group, medopar group and needling combined with medopar group was obviously decreased (P<0.05); there is no significant difference between needling group and nomal group; compared with medopar group and needling combined with medopar group, the expression in needling group increased significantly(P<0.05)。7. Effects of needling chorea trembling control area on the expression of Akt of PD mice.The result shows that in the area of substantia nigra, the expression in the needling group was higher than other four groups (P<0.05).There is no significant difference between other groups.In the CA1hippocampus area, the expression in the model group, needling group and needling combined with medopar group was obviously decreased compared with nomal group (P<0.05); the expression in the needling combined with medopar group increased compared with model group and medopar group in CA1, CA3and CA4area (P<0.05); compared with needling group, expression in needling combined with medopar group increased in CA1and CA4area (P<0.05).Conclusions:(1) Intraperitoneal injection of MPTP in C57BL/6J mice appears similar performance with PD patients in ethology and pathology, shows that we copy a PD model successfully. The decreased expression of BDNF, Trk B and PSD-95could have a connection with losing of TH positive neurons, but there is no significant evidence shows the connection between BDNF-Trk B pathway and PI3K/Akt pathway.(2) Needling the chorea trembling control area and combined with medopar treatment could have an effect in protecting DA neurons, and needling combined with medopar is better than needling use alone, while medopar use alone could not have an effect in DA neurons.(3) Needling the chorea trembling control area alone could have an effect in increasing the expression of Trk B and PSD-95rather than PI3K/Akt pathway.(4) Needling combined with medopar could increase the BDNF-Trk B and PI3K/Akt pathway in some degree to play a role of neuronprotective effect.(5) Medopar use alone could not have an effect through Trk B, PSD-95and PI3K/Akt pathway. |
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