益智宁神颗粒对SHR大鼠前额叶-纹状体多巴胺含量、GDNF mRNA及其蛋白表达的影响

Yizhi Ningshen granules: Effects on Dopamine levels, GDNFmRNA and its protein expression in prefrontal cortex-striatum of SHR

[目的]研究益智宁神颗粒对幼龄自发性高血压(SHR)大鼠前额叶-纹状体多巴胺含量、胶质细胞源性神经营养因子(GDNF)mRNA及其蛋白表达的影响。[方法]以32只幼年SHR大鼠为研究对象,采用随机区组法分为模型对照组、益智宁神组、盐酸哌甲酯组、托莫西汀组,另设8只京都(WKY)大鼠为正常对照组。按各组设计剂量连续给药4周后,取前额叶、纹状体,采用高效液相法检测多巴胺(DA)含量、逆转录-聚合酶链反应(RT-PCR)法检测GDNF mRNA表达、蛋白免疫印迹(Western blot)法检测GDNF蛋白表达。[结果]1)SHR各组大鼠前额叶、纹状体DA的含量均有低于WKY组的趋势,其中,益智宁神组大鼠前额叶DA含量明显高于模型对照组(P<0.05);益智宁神组大鼠纹状体DA含量低于正常对照组、盐酸托莫西汀组(P<0.05)。2)SHR各组前额叶、纹状体GDNF mRNA表达依次为盐酸托莫西汀组>盐酸哌甲酯组>益智宁神组>模型对照组(P<0.05)。3)SHR大鼠各组间前额叶、纹状体GDNF蛋白表达均高于WKY组;其中,益智宁神组前额叶GDNF蛋白表达与其余SHR大鼠各组无统计学差异(P>0.05),其纹状体GDNF蛋白表达低于盐酸哌甲酯组(P<0.05)。[结论]益智宁神颗粒能够提高SHR特定脑区DA含量、GDNF mRNA及其蛋白表达,对促进幼龄SHR大鼠特定脑区DA能神经网络成熟度具有一定的作用。

[Objective] In order to explain the mechanism of Chinese medicine compound granules, which named Yizhi Ningshen as the treatment of ADHD more than ten years, this study aimed to study the effects of Yizhi Ningshen granules on dopamine content, GDNFmRNA and protein expression in prefrontal and striatum of young SHR rats combined with modern molecular biology techniques. [Methods] Thirty-two young SHR rats were randomly divided into model group, Yizhi Ningshen group, methylphenidate hydrochloride group and atomoxetine group,8 WKY rats were randomly divided into normal control group. The rats were sacrificed for 4 weeks of continuous administration using designed doses, and the contents of dopamine in the prefrontal and striatum were detected by HPLC and the expression of GDNF mRNA was detected by RT-PCR, the expression of GDNF protein was detected by western blot. [Results] 1）The content of DA in the prefrontal cortex and striatum of SHR group was lower than that of WKY group, the content of DA in the prefrontal cortex was significantly higher than that in the model group（P0.05）. The content of DA in the striatum of the Yizhi Ningshen group was lower than that in the normal control group and tomoxetine group（P0.05）. 2）The expression of GDNF mRNA in the prefrontal cortex and striatum of SHR group from high to low order, the sequence is as follows, tomooxetine group methylenidate group Yizhi Ningshen group model group（P0.05）. 3）The expression of GDNF protein in the prefrontal cortex and striatum of each SHR group was higher than that in the WKY group, while the expression in striatu was lower than the methylphenidate group（P 0.05）. [Conclusion] Yizhi Ningshen granules can increase DA content and its specific nutrition factor GDNF mRNA and protein expression in specific brain regions of young SHR. It may play a role in promoting the maturation of dopaminergic neurons in specific brain regions of young SHR rats.