Dual Solid One Through The Needle In The Regulation Of Immune Function In Rat Model Of Parkinson's Disease

Objective:The Parkinson's disease (PD) is one of neural degeneration diseases whose characteristic is dopaminergic neuron proceeding degeneration and loss insubstantia nigra of midbrain. It predilection in middle-aged people and appears slow moving and progressivity in nervous system, the outbreak rate is placed in the second in neurodegenerative diseases which are 1%-3% over 65 years old of people. When the dopamine level reduced 70% to 80% in corpora striata, when the dopaminergic neuron degeneration of substantia nigra of midbrain reaches 50%, the functional disorder between catechol amine neurotransmitter and dopaminergic neuron degeneration appears more serious, which results the motor dysfunction of extracorticospinal tract and emerges some symptoms(static tremor, tetanize, movement brady, posture unsteadiness and so on)As for the cause of PD disease and outbreak mechanism are still not quite clarity, there wereexsitting these standpoints: heredity factor, environment factor, excitability nerve poison and calcic cell toxic action, oxidative stress, the age aging, mitochondria impairment, apoptosis and immunology abnormality. In the last few years, along with research thoroughly, scholars concerned the abnormality of the immunity gradually. Mattila report the immunology characteristic PD patients were the cellular immune function abnormality and the alteration of proportionality lymphocyte subgroups. Fiszer reported the cellular immune function degraded in PD patients, particularly in the activity IL-1. The IgA and IgM of humoral immunity increased, but not the IgG. Bongioanni discovered the IFN-γreceptors of high affinity in peripheral blood T cells were lower than health person and the TNF-αreceptors were higher than health person. All thesel explained the immunity network occurred derangement in the PD patients.There was still no special treatment of the PD. The total therapy principle is alleviativing symptoms and keeping the ability of self-care ability in early stage and maximatily extending the opening period, shorten the opening period and handling complications caused by medicines, symptom fluctuation and psychonosema in late stage. The medicine treatment or the surgical operation can't obstruct DA neurone continuesly; prevent the clinical symptoms aggravating proceeding. Acupuncture is to be used as a kind of special and safe nature therapy, which takes great effects on treating PD cases. In this research, we performed the method of "ShuangGuYiTong" electricity needle therapy to invest the mechanisms of action in regulating the immunity function in PD model and provide the new way and method of acupuncture therapy.Many researches shown that the gitter ceils may participate in the phases of occurrence and development in all of the central nervous system degenerative disease. The gitter cells activating were more important significance in the development of PD. First, the oxygen free radical damage may play the decisive significance in the dopaminergic neuron, impairment and the gitter cells activating is the main source of producing large amounts of free radicle; second, the histology researches expressed substantia nigra of midbrain is the most enriching district of gitter cells. In cerebrospinal fluid of PD sufferers, accompany with deprived dopaminergic neuron selectivity in substantia nigra of midbrain compact part, the gitter cells expressed human white cell antigen (HLA-DR) was activated. There was parallel relationship between the degrees of deprived dopaminergic neuron and the gitter ceils activating. The freed inflammatory cytokines and excessive free radicles after the gitter ceils activated mediated the damage of dopaminergic neuron. So if we inhibit the activation of gitter cells, we may reduce the loss of dopaminergic neuron maximatily.In recent years, there was much progress in the traditional Chinese medicine therapy curing the PD. Acupuncture as a kind of special, safety, convenient method was more used in clinical particularly. And it was obtained the affirmative curative effect. Milli-needles, head needles and electric needles were used in the treatment of PD. The puncture may affect the muscle potential, regulate the antioxidase system, improve the level of polyamine and changed the components of immune factor. But the thorough researches of mechanism of action relevant puncture were few. In the period work, the professor Wanghua thought that the main cause of PD lies in liver, the position of PDwere in the brain, definite the therapeutic principles of Tong tiao nao nuo, Xi feng zhi chart and Rou gan shu jin. According to the guidance of "ShuangguYitong" puncture, we choice the Guanyuan, Zusanli, Taichong and Fengfu four points to give electric acupuncture therapy in the mouse model of PD. And we discovered electric acupuncture could reinforce the TH activity in PD mouse; make the low expression of nitricoxide syntheses (NOS) and, aminoglutaminic acid (Glu) in corpora striata. In this study, we established the mouse middle-early model of PD and performed the methods of the immunology and the molecular biology to discuss the regulation function of acupuncture treatment to PD. And it may be established the foundation of revealing the function mechanism of acupuncture treatment to PD and provided some guidance for clinical treatment.Method:1. Establishment the early-middle partial side mouse PD in model: 80 SD Rats were divided into 4 groups: normal group, sham-modeling group, model group, and "ShuangGuYiTong" group. On rat brain stereotaxis instrument, 6-OH DA(10μg/5μL) were infused into corpora striata dorsal caudate putamen (CPU) of rats and apomorphine(0.5mg/kg) was peritoneal injection to inducing the rotational behavior after one week. If the average rotation speed were more than 7 rs/ min in 40 mins, the mold were established successly.2. "ShuangGuYiTong "acupuncture treatment method: Choosing Guanyuan, Zusanli, Taichong and Fengfu four points and the 30 type of acupuncture needle. After connecting the G6805 electricity needle treatment meter and adopting continuous wave, the condition were the frequency is 120 times/min, 1 time/day and every 30 minutes/time. The samples were taken after 30 days.3. "ShuangGuYiTong" acupuncture treatment affects to the gitter cells: Histochemical and immunohistochemical staining methods are adopted to observe the morphologic changes of substantia nigra and striatum and quantify the tyrosine hydroxylase (TH) positive neurons and nerve fibers by the detecting therapies of flow cytometry and immunohistochemistry; Adoption ricinus agglutinin marking gitter cells.4. "ShuangGuYiTong" acupuncture treatment affects to the expression of HLA-DR in PD model: The PER and the Western blotting technique detected the HLA-DR expression level.5. "ShuangGuYiTong" acupuncture treatment affects to IFN-γin lymphocytic subgroup and TNF-αreceptors. Using anti-cell factor receptors of monoclonal antibodies was to detect the level of IFN-γR and TNF-αR.6. "ShuangGuYiTong" acupuncture treatment affects to the level of IL-1 in PD model: the method of immunohistochemical was taken to invest it.7. "ShuangGuYiTong" acupuncture treatment affects to apoptosis of substantia nigra cells. The TH cell flow cytometry examination the apoptosis and PCR was used to detect the gene of Bax and Bcl-2 expression.Results:1. The total neurons of the substantia nigra pars compacta (SNc) of the destructive side, the total TH positive neurons and the striatum TH positive nerve fiber number in Model Group is obviously lower than that in normal group and sham-modeling group (P＜0.01). Compared to model group, the relevant indexes of the two needling groups are obviously increased (P＜0.01). The RAC positive of gitter cells almost didn't find in blank control group and sham operation group, but the number of gitter cells in the electricity needle treatment group were lower compared to model group in the same period (P＜0.05).2. The HLA-DRA, HLA-DRB1, 4 mRNA were high level expression in the model group than that of blank control group and sham-modeling group and electric acupuncture group. After the electric acupuncture treatment, HLA-DRA, HLA-DRB1,4 were lower obviously, but HLA-DRB3,5 had no this variety; HLA-DRA, HLA-DRB1,3,5 protein expressed high level in the model group than blank control group and sham-modeling group and electric acupuncture group. After the electric acupuncture treatment, they were low obviously.3. Compared the model group, blank control and sham operation group, the level of TNF-αR and IFN-γR in rat serum of electric acupuncture group reduced significantly (P＜0.01).4. The level ofIL-1βin model group was obviously high than that in groups of electric acupuncture, blank control and group sham operation.5. The apoptosis rate of substantia nigra cells in model group is the highest compared to other groups (P＜0.01), the apoptosis rate of substantia nigra cells in electric acupuncture group was low than model, it still high than control group and sham operation group (P＜0.01).6. The Bax mRNA expresses low level in substantia nigra and corpora striata of the model group especially in substantia nigra (P＜0.05). Compared to the model group, the Bax mRNA were more high expression, (P＜0.05), themRNAof Bcl-2 in the model group was only low expression in the substantia nigra than control group.Conclusions:1. Parkinson's model group exsits immunity abnormality, which can cause the changes of TNF-α, IFN-γ, IL-1βand HLA- DR.2. There have the changes of number of gitter cells.3. The ratio of apoptosis in cells of model group also increased.4. The electro-acupuncture method can regulate the PD immune function, increase the number of gitter cells and change its situation of apoptosis, which may be an effective method for theraping the PD.