帕金森病模型大鼠脑组织SPTLC2的表达研究

Study on SPTLC2 expression level in brain tissue in the rat model of Parkinson disease

目的探究丝氨酸棕榈酰转移酶长链亚基2(SPTLC2)在帕金森病大鼠脑组织中的表达。方法采用注射6-羟基多巴胺制备帕金森病大鼠模型,将模型组分为三组:模型组(2周)、模型组(4周)、模型组(8周),同时设置正常组,采用异常不自主动作(AIM)评分法评定各组大鼠行为,同时记录旋转持续时间、启动时间、剂峰旋转圈数。HE染色法观察大鼠脑组织变化,免疫组化法检测大鼠脑组织中α突触核蛋白的表达,逆转录聚合酶链反应(RT-PCR)法测定大鼠脑组织中SPTLC2 m RNA表达,Western blot检测大鼠脑组织中SPTLC2蛋白表达。结果模型组大鼠AIM评分、旋转持续时间、启动时间、剂峰旋转圈数均高于正常组,随着时间的延长,AIM评分、旋转启动时间、剂峰旋转圈数逐渐升高,旋转启动时间逐渐降低,差异有统计学意义(P<0.05)。免疫组化染色显示,正常组α突触核蛋白阳性细胞数量较少,模型组阳性细胞数量明显增多,且随着时间的延长,阳性细胞数量逐渐升高。与正常组、模型组(2周)相比,模型组(4周)、模型组(8周)SPTLC2基因、蛋白表达量均升高;与模型组(4周)相比,模型组(8周)SPTLC2基因、蛋白表达量升高,差异有统计学意义(P<0.05)。结论 SPTLC2可能参与帕金森病疾病的发生,通过上调SPTLC2基因、蛋白水平的表达来实现。

Objective To explore the expression of serine palmitate transferase long chain subunit 2（SPTLC2） in brain tissue in the rat model of Parkinson disease. Methods Parkinson rat model was prepared by injecting 6-hydroxy dopamine, and the rats were divided into three groups, 2-week model group, 4-week model group, 8-week model group, and normal group. Behaviors of rats in each group were assessed by using abnormal involuntary movement（AIM）scoring method. Rotation duration, starting time, the cycle number of agent peak were recorded meanwhile. HE staining method was used to observe the changes of brain tissue in rats, and the expression of α synuclein in rat brain tissue was detected by immunohistochemistry, and SPTLC2 and m RNA expression in brain tissue of rats was detected by using RT-PCR, and SPTLC2 protein expression in rat brain tissue was detected by Western blot. Results AIM score, rotation duration, starting time, agent peak rotation number in model group was higher than those in the normal group,and with the extension of time, AIM score, rotating start time, agent peak rotation number gradually increased, while the rotation starting time gradually decreased, with statistically significant differences（P〈0.05）. Immunohistochemical staining showed that the number of positive cells in the normal group was less than that in the normal group, and the number of positive cells in the model group increased significantly. The number of positive cells increased gradually with the extension of time. Compared with those in the normal group and 2-week model group, SPTLC2 gene and protein expression in 4-week and 8-week model group increased, and compared with 4-week model group, SPTLC2 gene and protein expression in 8-week model group increased, with statistically significant differences（P〈0.05）. Conclusion SPTLC2 may be involved in the pathogenesis of Parkinson disease, via up-regulating SPTLC2 gene and protein expression.