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Objective: Parkinson's disease (PD) is a common degenerative diseases of the central nervous system of unknown etiology and pathogenesis. The present study confirmed that inflammation plays an important role in PD etiology and pathogenesis. Activated microglia-mediated inflammatory reaction of DA neuronal injury with a start, activated microglia can release large amounts of proinflammatory cytokines, superoxide radicals, causing neurotrophic factor down mitochondrial dysfunction and oxidative stress, excitatory toxin-based cascade this pathological process for the the PD slow progress. Therefore inhibit inflammation, joint neuroprotective therapies to delay or even prevent the PD progress has become the new hotspot in the field. Selective COX-2 inhibitors - celecoxib celecoxib can block COX-2-mediated enzymatic reactions, inhibition of glial cell activation mainly a series of inflammatory reactions occur, thereby neuroprotective effect on DA neurons yuan. Ciliary neurotrophic factor (CNTF) promotes precursor neuronal growth, differentiation, maturation maintain the survival of neurons, and promote the regeneration of damaged neurons repair, prevent degeneration of neurons, and can act on astrocytes interstitial cells secrete a variety of neurotrophic factors to compensate for inadequate secretion of nerve factor. In this study, the cell culture model, select the inflammatory response inducer LPS selective COX-2 inhibitors celecoxib and ciliary neurotrophic factor of DA neurons co-cultured observed inflammatory response of DA neurons yuan of injury, and anti-inflammatory drugs and neurotrophic factors protective effect on DA neurons. For the ultimate cure for PD experimental basis. So far, the research at home and abroad rarely reported. Methods: SD rats were obtained from newborn within 24 hours, the ventral midbrain removed under sterile conditions, after digestion, centrifuged, resuspended and filtered into single cell suspension containing 2 × 106 / ml cells were seeded in bFGF20ng / ml, 2? 7 in DMEM/F12 culture medium, placed in 37 ° C, 5% CO2 humidified CO2 incubator culture. The 3d first half the amount vaccination medium was changed after the medium was changed every 2-3d full time. 7 days of in vitro culture, the cells covered the bottom of the bottle, synaptic fusion after LPS-induced inflammatory response made PD vitro model. The control group without LPS-induced and other intervening factors. Experimental group join LPS20ng/ml, mix well to make it fully contact, according to join the intervention factors were divided into four groups, (1) LPS group; (2) CNTF group; (3) CB group; (4) of CNTF CB group. The entire cell culture process inverted microscope under close observation of changes in cell morphology, and induction of 12h, 24h, respectively, detected in the culture supernatant of TNF-α concentrations (ELISA); immunocytochemistry assay time 24h 12h number of TH-positive cells were counted. Results: newborn mice within 24 hours of brain-derived nerve cells in vitro cultivation easy to survive, to maintain a longer period of time and in the role of neurotrophic factors. 24h culture seen most of the adherent cells, 48h some cells protrude 1-2 short protrusions, the cell monolayer distribution. 72h when the majority of the cells extended protrusions culture significantly increased 7 days after the cell body longer visible cell dendrites and axons, the formation of bipolar and multipolar morphology. ELISA assay culture supernatant TNF-α concentrations found that the content of TNF-α in the control group lower, higher levels of TNF-α in the experimental group, the same point in time in each experimental group compared to the control group, a significant difference (P lt; 0.01). The same point in time in each experimental group found that the lowest TNF-α content of CB of CNTF group (12h in 218.49 ± 15.78,24 h, 310.91 ± 25.69 pg / ml), followed by the order of the CB group of CNTF group, LPS group, the same point in time the difference between groups was significant (P lt; 0.01). Immunohistochemical detection of the control group, the experimental group, the number of TH-positive cells of the same, the difference was significant (P lt; 0.01) with the experimental group, the control group of the same point in time, compared to the same point in time in each experimental group, TH positive cells more CB CNTF group (12h was 50.65 ± 1.76,24 h 46.67 ± 1.62), followed by the CB group of CNTF group, LPS group, the difference between the same point in time group was significant (P lt; 0.01) the same experimental group different time points more found Paper No. CB of CNTF group, Paper No. CB group over time extend the TH-positive cell number decreased significantly, differences significant with (P lt; 0.05) in LPS group and CNTF group of TH-positive cell number to reduce more obvious difference is very significantly (P lt; 0.01). Conclusion: 24h within the neonatal rat brain-derived nerve cells were successfully cultured in vitro. Select inflammatory response inducing agent-LPS produced vitro PD inflammatory model. Confirmed the non-steroidal anti-inflammatory drugs - celecoxib, neurotrophin-CNTF can attenuate LPS on cultured DA neurons inflammatory injury, and celecoxib is superior of CNTF. Celecoxib cloth with CNTF Combined with stronger anti-inflammatory and neuroprotective effects.
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