The Effects Of Chinese Medicine Of Tonifying Kidney On MES23.5Dopaminergic Neuronal Cells Apoptosis

Objective:To investigate the effects of three different types of Chinese nourishing Kidney herbs Epimedium, Dodder and Herba through the observing of cell morphology and the testing of the apoptotic factors,NTFs in cell model of neuron apoptosis.Mothods:1. The experimental materials:The logarithmic growth phase of MES23.5dopaminergic cells are divided into six group s, including the blank control group, the model group, Epimedium group, Dodder group, Herba group,and Selegiline group. The blank control group were cultured by culture fluid containing25%of rat serum of the blank control group. The model group, Epimedium group, Dodder group, Herba group,and Selegiline group of MES23.5dopaminergic neuronal cells were treated with100umol/L H2O2in3hours.And then cultured by culture fluid containing25%of rat serum of the blank control group, the model group, Epimedium group, Dodder group, Herba group,and Selegiline group respectively。2.The detection contents:(1). Observation of cell morphology:observation of MES23.5dopaminergic neuronal cells morphology on light microscopy and electron microscopy.(2). Detection of apoptotic factors:the levels of caspase-3, Fas, Fas-L and bcl-2of MES23.5dopaminergic neuronal cells were detected by flow cytometry analysis;(3). Detection of NTFs:the levels of NGF,BDNF,GDNF of MES23.5dopaminergic neuronal cells were detected by immunohistochemistry analysis.Results:1. Comparison of cell morphology:(1) Observation of cell morphology on light microscopy:The cell membrane boundary of blank control group MES23.5dopaminergic neuronal cells were smooth and clear. And the soma were spindle-shaped,round or irregular in shape. And the cell body is full.The body of MES23.5dopaminergic neuronal cells in the model group turnned round and smaller. And observed cytoplasm of the model groups,the brightness increase, and visible circular translucent area could be found. the bouudary was not clear, and the cell appears to float off phenmenon of wall.A small amount of the cell in Epimedium group, Dodder group and Herba group can be s een apoptosis,and a little cell appears to float off phenmenon of wall,however most of neurons, morphology are normal compared with the blank control group.(2) Observation of cell morphology with on electron microscopy:The MES23.5dopamine nerve cells of blank control group are most of nearly round or oval. The membrane did not rupture or swell. There are some microvilli on the cell surface. The Nuclear is oval and mostly offset.The form and structure of the organelles is well.There is a little processes on the surface of MES23.5dopamine nerve cell in Model control group. There are some mitochondria in the cell, in which exist some of the ridge and the fractured or disappeared ridge is obvious.Rough endoplasmic is vesiculation and degranulation.There are some microvilli on the cell surface of MES23.5dopamine nerve cells in Epimedium and Herba group. The form of mitochondria is well. There are more mitochondria in the cell, in which exist most of the ridge and the fractured or disappeared ridge is not obvious.There is a little processes on the surface of MES23.5dopamine nerve cell in Dodder group. There are some mitochondria in the cell, in which exist some of the ridge and the fractured or disappeared ridge is obvious. Rough endoplasmic is vesiculation and degranulation.The MES23.5dopamine nerve cells of Selegiline group are most of nearly round or oval. The membrane did not rupture or swell. There are some microvilli on the cell surface. The Nuclear is oval and mostly offset. The form of mitochondria is well. There are more mitochondria in the cell, in which exist most of the ridge and the fractured or disappeared ridge is not obvious.2. Comparison of apoptotic factors: The levels of FasL in the model group were higher than the blank control group(p<0.05), Herba group and Selegiline group were lower than the model group(p<0.05); the levels of FasL had not existence obviously different among all groups(p>0.05); the levels of Caspase-3in the model group were higher than the blank control group, meanwhile Epimedium group, Herba group and Selegiline group were higher than the the model group(p <0.05); the levels of Bcl-2were obviously lower than the blank control group, Herba group were higher than the the model group(p<0.05).3. Comparison of NTFs:The levels of NGF were significantly lower in the model group than in blank control group (p<0.05), while the Herba group were higher than both the model group and selegiline group (p<0.05); the concentrations of BDNF in the model group were lower in blank control group,and both Dodder group and Herba group had higher levels than the model group and selegiline group (p<0.05); Epimedium group were higher than the model group (P<0.05); the levels of GDNF were significantly higher in the Herba group than in blank control group, model group and selegiline group (p<0.05); other subjects are not obviously different (P>0.05)Conclusion:Epimedium and Herba can modify apoptotic factors and NTFs in MES23.5dopaminergic neuronal cells to protect neuronal cells.lt is the possible mechanisms that Chinese nourishing Kidney herbs can delay the development of Parkinson.