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目的研究溶血磷脂酸(LPA)在大鼠体内对海马神经细胞tau蛋白磷酸化水平的影响和诱导神经细胞凋亡的细胞毒性作用。方法将72只SD大鼠分为实验组(n=32)、实验对照组(n=32)和对照组(n=8),利用脑立体定位技术在大鼠双侧海马微量注射溶血磷脂酸、溶剂,于注射后12、24、48和72h各不同时间点采用免疫组化方法测定该区域神经细胞中ser202位点磷酸化tau蛋白(PS202-tau)的表达,TUNEL技术检测细胞凋亡。结果实验组LPA注射后24h海马CA4区神经细胞中PS202-tau阳性表达到达高峰,阳性表达高于对照组和实验对照组(P<0.05)。LPA注射后48h TUNEL阳性细胞数达高峰,每个视野中阳性细胞数多于对照组和实验对照组(P<0.05)。结论LPA在动物整体水平可诱导大鼠海马神经细胞tau蛋白高度磷酸化,并导致神经细胞发生凋亡。
Objective To investigate the effect of LPA on the phosphorylation of tau in hippocampal neurons and the cytotoxicity induced by apoptosis in neurons. Methods 72 SD rats were divided into experimental group (n = 32), experimental control group (n = 32) and control group (n = 8). The brain stereotaxic technique was used to microinjection of lysophosphatidic acid , Solvent. The expression of phospho-tau protein (PS202-tau) at ser202 site in nerve cells in this region was detected by immunohistochemistry at different time points of 12, 24, 48 and 72 h after injection. Cell apoptosis was detected by TUNEL technique. Results The positive expression of PS202-tau in hippocampal CA4 neurons reached the peak at 24 h after LPA injection in experimental group, which was significantly higher than that in control group and experimental control group (P <0.05). At 48h after LPA injection, the number of TUNEL-positive cells peaked, and the number of positive cells per field was more than that of the control group and the experimental control group (P <0.05). Conclusion LPA can induce hyperphosphorylation of tau in hippocampal neurons and lead to neuronal apoptosis in the whole animal.