天麻素可改善多发性抽动症模型大鼠的头部抽动行为

Gastrodin can improve the head twitch behavior of rats with Tourette syndrome

目的探讨天麻素对多发性抽动症(TS)模型大鼠头部抽动行为的影响及相关机制。方法采用1-(2,5-二甲氧基-4-碘苯基)-2-氨基丙烷(DOI)诱导法制备TS大鼠模型,将40只Wistar大鼠随机均分为4组:正常组(生理盐水灌胃)、TS模型组(从造模首日起,注射DOI后2 h以生理盐水灌胃)、TS+天麻素组(从造模首日起,注射DOI后2 h以30 mg/kg天麻素灌胃);TS+天麻素+CHIR-99021[糖原合酶激酶-3β(GSK-3β)抑制剂]组(从造模首日起,注射DOI后2 h,先以2 mg/kg CHIR-99021腹腔注射,再以30 mg/kg天麻素灌胃),连续给药21 d。末次给药后,记录大鼠在30 min内的头部抽动次数,采用试剂盒检测纹状体5-羟色胺(5-HT)、5-羟吲哚乙酸(5-HIAA)、多巴胺(DA)水平,免疫组化法检测黑质中DA神经元数量,Western blot法检测纹状体中5-羟色胺转运蛋白(SERT)、5-羟色胺2A受体(5-HT2AR)、5-羟色胺2C受体(5-HT2CR)表达及GSK-3β磷酸化情况。结果与正常组比较,TS模型组大鼠头部抽动次数、DA水平、DA神经元数量、SERT水平及p-S9-GSK-3β、p-S9-GSK-3β/t-GSK-3β水平增高,5-HT水平降低(P<0.05)。与TS模型组比较,TS+天麻素组大鼠头部抽动次数、DA水平、DA神经元数量、SERT水平及p-S9-GSK-3β、p-S9-GSK-3β/t-GSK-3β水平降低,5-HT水平升高(P<0.05)。与TS+天麻素组比较,TS+天麻素+CHIR-99021组大鼠头部抽动次数、DA水平、DA神经元数量、SERT水平及p-S9-GSK-3β、p-S9-GSK-3β/t-GSK-3β水平升高,5-HT水平降低(P<0.05)。结论天麻素可改善TS大鼠头部抽动行为,其作用机制可能是通过抑制GSK-3β的磷酸化,调节SERT表达,影响5-HT水平,进而抑制纹状体中DA的释放。

Objective To investigate the effect and mechanism of gastrodin on head twitch behavior in rats with Tourette syndrome(TS).Methods 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane(DOI)induction method was used to replicate TS rat model.Forty Wistar rats were randomly divided into four groups(n=10):the normal group(gastric administration of normal saline),the TS model group(from the first day of modeling,intragastric administration with normal saline at 2 h after DOI injection),the TS+gastrodin group(from the first day of modeling,intragastric administration with 30 mg/kg gastrodin at 2 h after DOI injection),and the TS+gastrodin+CHIR-99021 group[glycogen synthase kinase-3(3(GSK-3(3)inhibitor](from the first day of modeling,2 h after DOI injection,intraperitoneal injection with 2 mg/kg CHIR-99021 firstly,and then intragastric administration with 30 mg/kg gastrodin),continuous administration for 21 d.After the last administration,the rat’s head twitching behavior within 30 min was counted.The levels of striatal 5-hydroxyindole(5-HT),5-hydroxyindole acetic acid(5-HIAA),and dopamine(DA)were detected by kits.The number of DA neurons in the substantia nigra was detected by immunohistochemical method.The expression of 5-HT transporter(SERT),5-hydroxyindole 2 A receptor(5-HT2 AR),5-hydroxyindole 2 C receptor(5-HT2 CR)and GSK-3βphosphorylation in the striatum were detected by Western blot assay.Results Compared with the normal group,the number of head twitches,DA level,number of DA neurons,SERT level and p-S9-GSK-3β,p-S9-GSK-3β/t-GSK-3βlevel increased in the TS model group,and the 5-HT level decreased(P<0.05).Compared with the TS model group,the number of head twitches,DA level,number of DA neurons,SERT level and p-S9-GSK-3β,p-S9-GSK-3β/t-GSK-3βlevel decreased in the TS+gastrodin group,and the 5-HT level increased(P<0.05).Compared with the TS+gastrodin group,the number of head twitches,DA level,number of DA neurons,SERT level and p-S9-GSK-3β,p-S9-GSK-3β/t-GSK-3βlevel increased,and the 5-HT level decreased in the TS+gastrodin+CHIR-99021 group(P<0.05).Conclusion Gastrodin can improve the head twitch behavior of TS rats,and its mechanism may be by inhibiting the phosphorylation of GSK-3β,regulating the expression of SERT,affecting the level of 5-HT,and then inhibiting the release of DA in striatum.