天麻钩藤饮对抽动障碍模型大鼠行为改善及神经保护作用

Behavior Improvement and Neuroprotective Effect of Tianma Gouteng Decoction on Tic Disorder Model Rats

关键词:  天麻钩藤饮  抽动障碍  多巴胺  PI3K/AKT/GSK-3β

Key words:  Tianma Gouteng decoction  tic disorder  dopamine  PI3K/AKT/GSK-3β

摘要:

目的 探究天麻钩藤饮对抽动障碍(tic disorder，TD)大鼠行为改善作用及其神经保护机制。方法 腹腔注射双(2-氰基乙基)胺诱导液进行大鼠TD造模，造模大鼠分为TD组，硫必利组(4.2 mg·kg^-1)，天麻钩藤饮低、中、高剂量组(19.6，39.2，78.4 g·kg^-1)，另将正常大鼠设为正常组，造模完成后观察大鼠行为能力。治疗4 周后，评价大鼠行为能力改善情况。分离各组大鼠脑部纹状体，采集血清，ELISA检测两者中多巴胺(dopamine，DA)、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白细胞介素1β(interleukin-1β，IL-1β)含量。HE染色观察大鼠纹状体病理改变情况，TUNEL染色观察纹状体内神经元凋亡情况，Western blotting测定大鼠纹状体内p-PI3K/PI3K、p-AKT/AKT、p-GSK-3β/GSK-3β蛋白表达情况。结果 治疗后，TD大鼠刻板行为、运动行为、旷场运动能力较治疗前均得到改善(P<0.01)。且与TD组相比，各组大鼠脑组织病理改变减轻；硫必利组和天麻钩藤饮中、高剂量组纹状体和血清中DA、TNF-α、IL-1β含量均降低(P<0.05或P<0.01)，纹状体内的神经元凋亡数量显著下降(P<0.05或P<0.01)，p-PI3K/PI3K、p-AKT/AKT蛋白表达均显著增加(P<0.05或P<0.01)；天麻钩藤饮中、高剂量组p-GSK-3β/GSK-3β蛋白表达均显著增加(P<0.05或P<0.01)。结论 天麻钩藤饮可改善TD大鼠行为，并可促进PI3K/Akt/GSK-3β信号通路磷酸化发挥神经保护作用，其作用效果具有剂量依赖性。

Abstract:

OBJECTIVE To explore the improvement of Tianma Gouteng decoction(TGD) on the behavior of tic disorder (TD) rats and its neuroprotective machenism. METHODS Iminodipropionitrile induction solution was injected intraperitoneally to establish TD rat model, then rats were divided into TD group, thiopride group(4.2 mg·kg^-1), TGD low, medium, high dose group(19.6, 39.2, 78.4 g·kg^-1), normal rats were set into normal group, behavioral capacity of rats were measured after modeling. After 4-week treatment, behavioral capacity improvement of rats was evaluated. The striatum of rats in each group was isolated, and serum was collected, ELISA was used to detect the contents of dopamine(DA), tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β). HE staining was used to observe the pathological changes in the rat striatum, TUNEL staining was used to observe the neuronal apoptosis in the rat’s striatum, and Western blotting was used to determine the protein expressions of p-PI3K/PI3K, p-AKT/AKT, and p-GSK-3β/GSK-3β in the rats’t striatum. RESULTS After treatment, the stereotyped behavior, motor behavior and exercise ability of the TD rats were improved compared to those before treatment (P<0.01). And compared with TD group, pathological changes in each group were reduced; and content of DA, TNF-α, IL-1β in thiopride group and TGD medium, high-dose groups were significantly decreased(P<0.05 or P<0.01), the number of apoptotic neurons of striatum significantly decreased(P<0.05 or P<0.01), and the protein expressions of p-PI3K/PI3K and p-AKT/AKT were significantly increased(P<0.05 or P<0.01); the protein expressions of p-GSK-3β/GSK-3β in TGD medium, high-dose groups were significantly decreased(P<0.05 or P<0.01). CONCLUSION TGD improves the behavior of TD rats, and promotes the phosphorylation of PI3K/Akt/GSK-3β signaling pathway to exert neuroprotective effect, and its effect is dose-dependent.