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目的观察乙醇处理大鼠脑内酪氨酸羟化酶(TH)和多巴胺转运体(DAT)的表达变化,判断乙醇对脑内多巴胺(DA)能体系的影响。方法选取Wistar大鼠60只,随机分为对照组和乙醇处理组,每组30只,乙醇处理组大鼠以20%乙醇代替饮水饲养6个月;利用免疫组织化学、流式细胞术及免疫印迹等方法,分析乙醇处理大鼠有关脑区TH和DAT的表达改变。结果1.免疫细胞化学法研究发现,乙醇处理组大鼠脑内黑质(SN)-腹侧被盖区(VTA)、尾壳核(CPu)和伏核(NACC)TH的灰度值明显低于对照组(P<0.05);尾壳核和伏核DAT的灰度值明显低于对照组(P<0.05)。2.流式细胞术检测发现,乙醇处理组大鼠中脑TH的表达量明显高于对照组(P<0.05)。3.免疫印迹检测发现,乙醇处理组大鼠所观察脑区TH和DAT与β-actin条带相对吸光度比值明显高于对照组(P<0.05)。结论乙醇处理增加大鼠脑内TH和DAT的表达。
Objective To observe the expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in brain of ethanol-treated rats and to determine the effect of ethanol on dopamine (DA) energy in the brain. Methods Sixty Wistar rats were randomly divided into control group and ethanol group, with 30 rats in each group. Rats in ethanol group were fed with 20% ethanol instead of drinking water for 6 months. Immunohistochemistry, flow cytometry and immunization Blot and other methods to analyze ethanol-related brain regions TH and DAT expression changes. Immunocytochemistry showed that gray value of SN, ventral tegmental area (VTA), caudate putamen (CPu) and nucleus accumbens (NACC) TH in ethanol-treated rats were significantly (P <0.05). The gray value of DAT in caudate putamen and nucleus accumbens was significantly lower than that in control group (P <0.05). The results of flow cytometry showed that the expression of TH in the midbrain of ethanol-treated rats was significantly higher than that of the control group (P <0.05). Western blotting showed that the ratio of relative absorbance of TH and DAT to β-actin bands in the ethanol-treated rats was significantly higher than that of the control group (P <0.05). Conclusion Ethanol treatment can increase the expression of TH and DAT in rat brain.