塞来昔布防护DA能神经元损伤实验研究

The study on the neuroprotective effects of celecoxib against degeneration of dopaminergic neurons

目的观察非甾体类抗炎药物-塞来昔布对DA能(DA)神经元损伤的保护作用。方法取新生SD大鼠进行中脑神经细胞的原代培养。采用脂多糖(LPS)制作体外PD炎症模型。实验分为对照组、LPS组、塞来昔布(CB)组。倒置显微镜下观察细胞形态变化,第12小时、第24小时测定中脑细胞培养液上清TNF-α浓度及进行酪氨酸羟化酶(TH)免疫细胞化学检测。结果倒置显微镜下见,12h时LPS组细胞体积略有肿胀,折光性增强,突起变细、缩短,胶质细胞活化。24h时细胞数目明显减少,胞体皱缩,突起明显变细、缩短,部分细胞甚至崩解。对照组、CB组细胞形态变化不明显。12hLPS、CB组中脑神经细胞培养液上清TNF-α浓度与对照组相比均显著升高(P<0.01)。组间比较,LPS组TNF-α浓度比CB组明显升高(P<0.01)。24h时实验两组TNF-α浓度与12h时相比均显著增高(P<0.01)。对照组TH阳性细胞数较多。LPS组部分神经元衰亡,12hTH阳性细胞数与对照组相比显著减少,24h减少更明显(P<0.01)。CB组TH阳性细胞数与对照组相比亦显著减少(P<0.01)。组间比较,LPS组TH阳性细胞数与CB组相比明显减少。结论塞来昔布可减轻LPS对体外培养DA能神经元的炎性损伤作用。

Objective To observe the neuroprotective effects of celecoxib against degeneration of dopaminergic neurons.Methods The primary mesencephalic neural cell deriving from SD newborn rat were cultured.Lipopolysaccharide （LPS） were used to establish inflammator reaction model of PD in vitro.Cultured cells were divided into control group,LPS and CB groups.The concentration of tumor necrosis factor-α（TNF-α） in the culture medium were determined by using ELISA method,and tyrosine hydroxylase immunohistochmisry were performed after cutured for 12 hours and 24 hours. Results The concentration of TNF-α in control group was remarkably lower than in experiment groups at the same time point. （ P 〈0.01）.The concentration of TNF-α in LPS group was highest, respectively：in CB group. The amount of TH-positive cells in control group were more than in experimental groups（ P 〈0.01）. The amount of TH-positive cells in LPS group were lowerest,respectively：in CB group.Conclusion Celecoxib have neuroprotective effect through lessening the degeneration of dopaminergic neurons caused by LPS in vitro.