| Parkinson's disease (PD) is a kind of chronic neurodegenerative disease, The main clinical manifestations are tremor,rigidity and Bradykinesia. PD' Pathogenesis is related to loss of dopaminergic neurons and striatal dopamine. Further studies show that PD is related to brain mitochondrial dysfunction. Currently, there is no any ideal method of treatment of PD. Acupuncture has achieved significant effect and it is gradually understood and used by people. But the study of mechanism about Acupuncture treatment of PD is inadequate. Therefore, in this study, we will do a further research about treatment of PD on the basis of PD model etablished,by acupuncturing Du points. We will observe protective effect of acupuncture on dopaminergic neurons and mitochondrial complex by acupuncturing Du points by many kinds of techniques such as behavioral test, high pressure liquid chromatograph, immunohistochemistry, biochemistry, electron microscopy etc. We analyse the mechanism of acupuncture to improve mitochondrial function. So we can provide a theoretical basis for clinical treatment of acupuncture.This study is consisted of 2 parts:(1) By observing the influence of acupuncture on PD mice's behavior, dopaminergic neurons, dopamine in the striatum and its metabolite dihydroxyphenylacetic acid by acupuncturing Dazhui and Baihui,we will study protective effects of acupuncturing Du points on dopaminergic neurons. (2) On the basis of the first experiment, we will further observe the influence of acupuncture on activities of mitochondrial complexⅠ-Ⅳof PD mice, so as to study the mechanism of acupuncture to protect mitochondrion function.The establ ishment of animal models and grouping:We set up PD models by using C57BL/6 mice,doing intraperitoneal injection by using 0.3%MPTP (prepared with saline), continuous injecting for 5 days. The animals were randomly divided into 5 groups, such as normal group, model group, acupuncture group, madopar group and acupuncture mixed madopar group. Mice were killed after 14 days,28 days and 56 days treatment with MPTP.The main experimental results are as follows:(1)The effect of acupuncture on behavior of PD mice.After intraperitoneal injection of MPTP, there are different levels of movement disorder in mice. By the 14th day after modeling, as for the mice of model group, the climbing pole time was significantly longer (p＜0.05), the time of acupuncture group and Acupuncture-Madopar group are short than model group(p＜0.05). But the climbing pole time of the mice in madopar group were not significantly shorter. By the 28th day after modeling, The climbing pole time of the mice in the three treatment group were all shorter than model group. By the 56th day after modeling, the climbing pole time of the mice in all groups had no significant difference.(2)The effect of acupuncture on the number of TH-positive neuronsAs for the animals of 14-day survivaltime,in substantia nigra of model group, TH-positive cell bodies and fibers are sparse, the number of neurons are reduced. In three treatment groups, the number of TH-positive neurons are more than model group. As for the animals of 28-day survivaltime, the number of neurons in model group are much less than normal group (p＜0.05). The number of neurons in three treatment group are also less than normal group (p＜0.05). The number in acupuncture-madopar group are more than madopar group (p＜0.05). As for the animals of 28-day survivaltime, the number of neurons in model group are much less than normal group(p＜0.05). The number of neurons in acupuncture group and acupuncture-madopar group are obviously more than model group(p＜0.05).(3)The effect of acupuncture on DA and its metabolites DOPAC.In 14-day survivaltime, compared with normal group, brain DA and DOPAC in model groups is obviously reduced (p＜0.05), brain DA and DOPAC of three treatment groups are not significantly more than the model group. In 28-day survivaltime, brain DA in model groups is obviously reduced(p＜0.05),the content of DA three treatment groups are not different with model group. The DOPAC in acupuncture-madopar group are more than model group(p＜0.05).(4)The effect of acupuncture on ultrastructure of brain mitochondria of PD miceIn the three survivaltimes, mitochondria disorder or disappear and lysosomes increase in model group. In 14-day survivaltime, Nucleus membrane invagination less in three treatments than model group. In 28-day survivaltime, in acupuncture group and acupuncture-madopar group, a little mitochondria disappear, nucleus membrane structure is clear. In 56-day survivaltime, some mitochondria in acupuncture group disapear,little nucleus membrane structure in madopar group invaginate, nucleus membrane structure is clear in acupuncture-madopar group.(5)The effect of acupuncture on brain mitochondrial complex of PD mice①The effect of acupuncture on brain mitochondrial complexⅠThe mitochondrial complexⅠ's activity of model groups significantly descend in 14-day survivaltime and 56-day survival time(p＜0.05). The mitochondrial complexⅠ's function of medopar groups significantly also descend compared to normal groups in 14-day survivaltime and 56-day survivaltime. However, the function of acupuncture group and acupuncture-madopar group have no obvious descent (p＜0.05).①The effect of acupuncture on brain mitochondrial complexⅡcomplexⅡ's activity of model group in 56-day survivaltime obvious descend(p＜0.05). Acupuncture groups in 14 and 56-day survivaltimes have significantly high complexⅡ's fuction. The other two treatment groups are not different with normal and model groups (p＞0.05).③The effect of acupuncture on brain mitochondrial complexⅢIn 14 and 28-day survivaltimes,all groups are not different. In 56-day survivaltime, the complexⅢ's activity of model group is lower than normal group(p＜0.05), the fuction of acupuncture group and acupuncture-madopar group are very higher than model group (p＜0.05), the fuction of madopar group and madopar group are lower than acupuncture group (p＜0.05).④The effect of acupuncture on brain mitochondrial complexⅣIn 14-day survivaltime, all groups are not different. In 28-day survivaltime, the complexⅣ's activity of model group is lower than normal group (p＜0.05), t he fuction of acupuncture group and acupuncture-madopar group are very higher than model group(p＜0.05), In 56-day survivaltime, the fuction of model group is lower than normal group(p＜0.05), the fuction of madopar group and acupuncture-madopar group are very higher than model group (p＜0.05).Conclusion:The experimental results suggest that acupuncture or acupuncture-madopar could adjust Du point and reduce the tremor to some extent. The mechanism may be that it can reduce the damage and loss of dopaminergic neurons and increase mitochondrial complex activities of PD mice. So the study may provide modern scientific basis for the application of acupuncture in PD' s treatment. |
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