孔圣枕中丹对自发性高血压大鼠前额叶皮质及纹状体TH、DAT、DRD1、DRD2蛋白和基因表达的影响

目的探讨孔圣枕中丹对注意缺陷多动障碍（ADHD）动物模型——自发性高血压大鼠（SHR）前额叶皮质及纹状体（含伏核）中酪氨酸羟化酶（TH）、多巴胺转运体（DAT）、多巴胺D1受体（DRD1）、多巴胺D2受体（DRD2）蛋白和基因表达的影响。方法将36只SHR雄性大鼠随机分为三组,分别用孔圣枕中丹煎剂、盐酸哌甲酯和生理盐水灌胃28 d,采用免疫组化法和原位杂交法检测其前额叶皮质、纹状体（含伏核）中TH、DAT、DAR1、DRD2蛋白和基因的表达。结果①在SHR大鼠的前额叶皮质：TH的蛋白表达,孔圣枕中丹组（10.10±3.25）高于盐酸哌甲酯组（7.78±2.73）,后者又高于生理盐水对照组（4.79±2.53）;TH的基因表达,孔圣枕中丹组（23.00±2.10）高于盐酸哌甲酯组（19.26±1.26）,后者又高于生理盐水对照组（16.63±3.09）;DAT的蛋白表达,孔圣枕中丹组（9.06±1.81）低于盐酸哌甲酯组（14.20±1.19）,后者又低于生理盐水对照组（17.57±3.74）;DAT的基因表达,孔圣枕中丹组（9.32±1.40）低于盐酸哌甲酯组（16.45±3.23）,后者又低于生理盐水对照组（26.28±4.48）。DRD1的蛋白表达,孔圣枕中丹组（14.67±1.49）和盐酸哌甲酯组（11.45±1.42）均低于生理盐水对照组（18.06±2.20）;DRD1的基因表达,孔圣枕中丹组（22.54±2.69）和盐酸哌甲酯组（16.47±1.25）均低于生理盐水对照组（38.65±1.10）。DRD2的蛋白表达,孔圣枕中丹组（14.83±1.84）和盐酸哌甲酯组（11.01±1.03）均低于生理盐水对照组（17.83±3.36）;DRD2的基因表达,孔圣枕中丹组（14.68±1.03）和盐酸哌甲酯组（11.18±1.01）均低于生理盐水对照组（17.44±2.84）。②在SHR大鼠的纹状体：TH的蛋白表达,孔圣枕中丹组（14.30±3.85）高于盐酸哌甲酯组（9.36±2.59）,后者又高于生理盐水对照组（6.30±3.21）;TH的基因表达,孔圣枕中丹组（22.92±1.87）高于盐酸哌甲酯组（19.24±0.77）,后者又高于生理盐水对照组（16.47±1.19）。DAT的蛋白表达,孔圣枕中丹组（9.78±1.24）低于盐酸哌甲酯组（14.30±2.43）,后者又低于生理盐水对照组（17.79±2.39）;DAT的基因表达,孔圣枕中丹组（11.75±1.99）低于盐酸哌甲酯组（17.72±2.77）,盐酸哌甲酯组又低于生理盐水对照组（30.54±9.08）。DRD1的蛋白表达,孔圣枕中丹组（14.24±2.19）和盐酸哌甲酯（11.22±2.78）组均低于生理盐水对照组（22.10±2.90）;DRD1的基因表达,孔圣枕中丹组（17.29±1.27）和盐酸哌甲酯组（14.02±1.55）均低于生理盐水对照组（23.12±2.70）。DRD2的蛋白表达,孔圣枕中丹组（11.05±1.39）和盐酸哌甲酯组（13.83±1.20）均低于生理盐水对照组（16.64±0.72）;DRD2的基因表达,孔圣枕中丹组（9.68±1.65）和盐酸哌甲酯组（12.70±0.70）均低于生理盐水对照组（17.15±1.05）。以上差异均有统计学意义（均P〈0.05）。结论孔圣枕中丹可能通过影响调控脑内前额叶皮质-纹状体环路的DA神经信号传导而治疗ADHD。

Effect of Kongsheng Zhenzhong Pill on protein and gene expression of TH,DAT,DRD1,DRD2 in the prefrontal cortex and striatum of spontaneous hypertensive rat

Objective To observe the effects of Kongsheng Zhenzhong Pill on protein and gene expression of TH,DAT,DRD1,DRD2 in the prefrontal cortex and striatum of SHR.Methods 36 spontaneous hypertensive rats（SHR） were randomly divided into Kongsheng Zhenzhong Pill group,Methylphenidate Hydrochloride group,and saline control group,KongshengZhenzhongPillelixation,Methylphenidate Hydrochloride and saline were administered respectively to 3 groups by average.The protein and gene expression of TH,DAT,DRD1,DRD2 in the prefrontal cortex and striatum of SHR were detected 28 days later by immunohisto chemistry and in situ hybridization.Results ①In the prefrontal cortex： the TH protein expression level of Kongsheng Zhenzhong Pill group（10.10±3.25） were higher than that of Methylphenidate Hydrochloride group（7.78 ±2.73）,which were higher than that of saline control group（4.79 ±2.53）.The gene expression levels of three groups above were（23.00±2.10）,（19.26±1.26） and（16.63±3.09） respectively,Kongsheng Zhenzhong Pill group were the highest.The DAT protein expression level of Kongsheng Zhenzhong Pill group（9.06±1.81） were lower than that of Methylphenidate Hydrochloride group（14.20±1.19）,which were lower than that of saline control group（17.57±3.74）.The gene expression levels of three groups above were（9.32±1.40）,（16.45±3.23） and（26.28±4.48） respectively,Kongsheng Zhenzhong Pill group was the lowest.The DRD1 protein expression levels of Kongsheng Zhenzhong Pill group（14.67 ±1.49） and Methylphenidate Hydrochloride group（11.45±1.42） were lower than that of saline control group（18.06 ±2.20）.The DRD1 gene expression levels of Kongsheng Zhenzhong Pill group（22.54±2.69） and Methylphenidate Hydrochloride group（16.47±1.25） were also lower than that of saline control group（38.65±1.10）.The DRD2 protein expression levels of Kongsheng Zhenzhong Pill group（14.83±1.84） and Methylphenidate Hydrochloride group（11.01 ±1.03） were lower than that of saline control group（17.83 ±3.36）.The DRD2 gene expression levels of Kongsheng Zhenzhong Pill group（14.68 ±1.03） and Methylphenidate Hydrochloride group（11.18±1.01） were also lower than that of saline control group（17.44±2.84）.②In the striatum： the TH protein expression level of Kongsheng Zhenzhong Pill group（14.30 ±3.85） were higher than that of Methylphenidate Hydrochloride group（9.36±2.59）,which were higher than that of saline control group（6.30±3.21）.The gene expression levels of three groups above were（22.92±1.87）,（19.24±0.77） and（16.47±1.19） respectively,Kongsheng Zhenzhong Pill group were the highest.The DAT protein expression level of Kongsheng Zhenzhong Pill group（9.78±1.24） were lower than that of Methylphenidate Hydrochloride group（14.30 ±2.43）,which were lower than that of saline control group（17.79 ±2.39）.The gene expression levels of three groups above were（11.75 ±1.99）,（17.72 ±2.77） and（30.54±9.08） respectively,Kongsheng Zhenzhong Pill group was the lowest.The DRD1 protein expression levels of Kongsheng Zhenzhong Pill group（14.24±2.19） and Methylphenidate Hydrochloride group（11.22 ±2.78） were lower than that of saline control group（22.10 ±2.90）.The DRD1 gene expression levels of Kongsheng Zhenzhong Pill group（17.29±1.27） and Methylphenidate Hydrochloride group（14.02±1.55） were also lower than that of saline control group（23.12 ±2.70）.The DRD2 protein expression levels of Kongsheng Zhenzhong Pill group（11.05 ±1.39） and Methylphenidate Hydrochloride group（13.83 ±1.20） were lower than that of saline control group（16.64 ±0.72）.The DRD2 gene expression levels of Kongsheng Zhenzhong Pill group（9.68 ±1.65） and Methylphenidate Hydrochloride group（12.70±0.70） were also lower than that of saline control group（17.15 ±1.05）.The differences above were all significant（all P 0.05）.Conclusion Kongsheng Zhenzhong Pill may affect the transmission of dopamine which modulat ing prefrontal cortex-striatum loop.