健脾止动汤对多发性抽动症模型大鼠组胺及组胺脱羧酶的影响

Effect of Jianpi Zhidong Decoction on the Expression of Histamine and Histamine Decarboxylase in Tourette Syndrome Model Rats

目的:探讨健脾止动汤对多发性抽动症(TS)模型大鼠纹状体及血清组胺(HA)、组胺脱羧酶(HDC)表达的影响。方法:将24只SD大鼠按照随机数字表法分为空白组、模型组、中药组和西药组,每组6只。除空白组外,其他组均给予250 mg/kg IDPN腹腔注射7 d构建TS模型。空白组、模型组用15 mL/kg生理盐水灌胃,中药组用16 g/kg健脾止动汤灌胃,西药组给予21 mg/kg盐酸硫必利灌胃,1次/d,连续干预6周。对比各组大鼠体质量、行为学变化。用酶联免疫吸附试验(ELISA)检测大鼠纹状体及血清HA,用反转录聚合酶链反应(RT-PCR)检测大鼠纹状体HDC mRNA表达,用蛋白质印迹法和免疫组织化学法检测大鼠纹状体HDC蛋白表达。结果:模型组、健脾止动汤组、西药组体质量比较差异无统计学意义(P>0.05);干预6周,与模型组比较,健脾止动汤组、西药组体质量均低(P<0.05)。模型组、健脾止动汤组、西药组行为学评分比较差异无统计学意义(P>0.05);干预6周,与模型组比较,健脾止动汤组、西药组行为学评分均低(P<0.01)。与空白组比较,模型组大鼠血清HA水平低(P<0.05);与模型组比较,健脾止动汤组大鼠血清HA水平高(P<0.05),西药组血清HA水平高,但差异无统计学意义(P>0.05)。与空白组比较,模型组大鼠纹状体HA水平低(P<0.05);与模型组比较,健脾止动汤组及西药组大鼠纹状体HA水平高(P<0.01)。与空白组比较,模型组纹状体HDC mRNA表达量低,但差异无统计学意义(P>0.05);与模型组比较,健脾止动汤组及西药组大鼠纹状体HDC mRNA表达量高,但差异无统计学意义(P>0.05)。蛋白质印迹法检测结果:与空白组比较,模型组大鼠纹状体HDC蛋白表达低(P<0.05);与模型组比较,健脾止动汤组纹状体HDC蛋白表达高(P<0.05),西药组纹状体HDC蛋白表达高,但差异无统计学意义(P>0.05)。免疫组织化学法检测结果:与空白组比较,模型组纹状体组胺脱羧酶IOD值低(P<0.05);与模型组比较,健脾止动汤组纹状体HDC IOD值高(P<0.05),西药组HDC IOD值高,但差异无统计学意义(P>0.05)。结论:健脾止动汤可提高TS大鼠血清、纹状体HA水平及纹状体HDC表达。

Objective:To investigate the effect of Jianpi Zhidong Decoction on the histamine(HA)and histidine decarboxylase(HDC)expression in the striatum and serum in a Tourette syndrome(TS)rat model.Methods:Twenty-four SD rats were randomly assigned to blank,model,Chinese medicine,and western medicine groups according to a random number table,with six rats in each group.Except for the blank group,all other groups were given IDPN at 250 mg/kg via intraperitoneal injection for 7 days to establish the TS model.The blank and model groups were administered with saline at 15 mL/kg,the Chinese medicine group with Jianpi Zhidong Decoction at 16 g/kg,and the western medicine group with sulpiride at 21 mg/kg,once daily,for 6 weeks.Body weight and behavioral changes of all groups were compared.Enzyme-linked immunosorbent assay(ELISA)was used to measure HA levels in the striatum and serum.Reverse transcription polymerase chain reaction(RT-PCR)was used to detect HDC mRNA expression in the striatum.Western blot and immunohistochemistry were used to assess HDC protein expression in the striatum.Results:There were no significant differences in body weight among the model,Jianpi Zhidong Decoction,and western medicine groups(P>0.05).After 6 weeks of intervention,both the Jianpi Zhidong Decoction and western medicine groups had lower body weight than the model group(P<0.05).Behavioral scores showed no significant differences between the model,Jianpi Zhidong Decoction,and western medicine groups(P>0.05).After 6 weeks of intervention,the Jianpi Zhidong Decoction and western medicine groups had lower scores than the model group(P<0.01).Compared with the blank group,the model group showed lower serum HA levels(P<0.05).However,serum HA levels were higher in the Jianpi Zhidong Decoction group than in the model group(P<0.05),while the western medicine group showed a non-significant increase(P>0.05).Striatal HA levels were lower in the model group than in the blank group(P<0.05),but higher in the Jianpi Zhidong Decoction and western medicine groups than in the model group(P<0.01).Striatal HDC mRNA expression was lower in the model group than in the blank group but the difference was not significant(P>0.05).Compared with the model group,the Jianpi Zhidong Decoction and western medicine groups showed the higher striatal HDC mRNA expression,but the difference was not significant(P>0.05).Western blot results indicated that striatal HDC protein expression was lower in the model group than in the blank group(P<0.05),while it was higher in the Jianpi Zhidong Decoction group than in the model group(P<0.05)and the western medicine group(P>0.05).Immunohistochemistry showed that the IOD value of HDC in the striatum was lower in the model group than in the blank group(P<0.05).Compared with the model group,the IOD value of HDC in the striatum was higher in the Jianpi Zhidong Decoction group(P<0.05)and the western medicine group(P>0.05).Conclusion:Jianpi Zhidong Decoction can increase serum and striatal HA levels and striatal HDC expression in TS rats.