| Background:Parkinson's disease, a common degenerative disease of the central nervous system, which basic pathological change is be divided into six stage by Braak,that begins from the medulla pons, midbrain, and finally the brain and the neocortex. The rate of disease incidence unceasingly grows along with the advance of age obviously. In our country,every five hundred in one hundred thousand would have parkinson's disease between thirty and sixty yesrs old,above sixty, every one thousand in one hundred thousand with the ration of male to female is 2-3:1,above fifty five years old,the incidence is one point nought seven percent,while above seventy five years old,is two point five percent. These data is close with that reported in 2005 in European and American countries, and was obviously higher than that reported in the 1980s in our country's survey result by far. PD is affecting senior citizen's quality of life seriously. At present there is no effective method or the medicine that discovered can cure the PD patient thoroughly. Erythropoietin, is one kind the glycoprotein hormone which participates in the adjustment of differentiation, multiplication, suppressing perishes of red blood cell precursor. Exogenetic EPO experimental study discover that EPO can reduce the neuron damage in brain and spinal cord's ischemia and the flesh wound, under the status epilepticus, the subarachnoid hemorrhage and experimental auto immunity encephalomyelitis,which demonstrated that EPO has widespread nerve protective function. At present the range of study is restricted in the protection of EPO on the acute brain damage,specially in the protection on the ischemia brain damage. In our study,Parkinson's disease cell model.was built by 6- OHDA,and in the model the influence of EPO to the dopaminergic neuron was Observed.Objective:To investigate the protection and its mechanism of EPO upon dopaminergic neuron in Parkinson's disease cell models induced by 6-OHDA.Methods:Culture mesencephalon original generation cells of pregnant 13-14 day of SD rats, which were divided into four groups:(1)control group, original generation cells were cultured; (2) 6-OHDA group, original generation cells with uric acid; (3) 6-OHDA+EPO group:(group A-F),each group of different concentration EPO from 0.1,1,3,10 to 20 U/ml, (4) 6-OHDA+EPO+AG490:group E was 10 U/ml EPO+10 U/ml AG490.Add different concentration EPO in the group till day 6,till day 10 group A,B,C,D,F was added in 6-OHDA (100μmol/L),meanwhile,group E was added AG490 for one hour, cells were collected and flow cytometry wes used, using Rh123 to examine the plastochondria membrane potential, immunohis-tochemistry to delect the expression of tyrosine hydroxylase (TH) and caspase-3. Results:Compared with 6-OHDA group, The TH positive cell number increased obviously in group B=,C,D,F(P<0.01), on the contrary, caspase-3 positive cell number decreased obviously in group B,C,D,E,F(P<0.01),compared with group A,B,C,E,F, in group D, TH positive cell number increased obviously,while caspase-3 positive cell number decreased obviously, compared with group E, TH positive cell number increased obviously in group C,D,F, caspase-3 positive cell number decreased obviously in group B,C,D,F(P<0.01). Compared with the control group, plastochondria membrane potential decreased distinctly in 6-OHDA group (P<0.01),while increased clearly in group A,B,C,D,F (P<0.01),between group A to F, plastochondria membrane potential increased distinctly in group D (P<0.01), meanwhile decreased distinctly in group E (P<0.01)Conclusions:Dopaminergic neurons were pretreated by EPO in Vitro Parkinson's disease models, which can decrease the damge of dopaminergic neurons, stabilize the plastochondria membrane potential; and the mechanism may be related to its neuroprotective property by reducing the oxidative stress to decrease dopaminergic neuronal apoptosis,thus to protect the dopaminergic neurons. |
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