安君宁对吗啡依赖大鼠脑内酪胺酸羟化酶和胶质原纤维酸性蛋白表达的影响

The effects of ANJUNNING on tyrosine hydroxylase(TH) and glial fibrillary acidic protein(GFAP) in VTA of morphine dependent SD rats

目的 观察安君宁对吗啡依赖大鼠腹侧被盖区（VTA）内酪胺酸羟化酶（TH）和胶质原纤维酸性蛋白（GFAP）表达的影响.方法 选择体质量在180～220g雄性SD大鼠30只为研究对象,动物随机分为5组（每组动物6只）,即单用生理盐水正常对照组（Normal组）、其余四组大鼠则先制成吗啡成瘾模型：以5mg/kg/次开始,每天增加5mg,一直增加到第10天50mg·kg^-1·次^-1,每天腹腔注射吗啡2次.第11天开始自然戒断,四组成瘾大鼠随机分成吗啡成瘾后戒断并安君宁治疗12d组（P12E）、吗啡成瘾后戒断并淀粉治疗12d组（P12EC）,吗啡成瘾后戒断并安君宁治疗30d组（P30E）、吗啡成瘾后戒断并淀粉治疗30d组（P30EC）.安君宁和淀粉均为每天灌胃2次,安君宁剂量为3 g·kg^-1·次^-1,对照组喂等量淀粉.用ABC法测VTA的TH与GFAP的免疫组织化学反应;测定阳性神经元的平均光密度（OD）值.结果 5组大鼠（Normal、P12EC、P12E、P30EC、P30E组）VTA中TH免疫反应的强度分别为（0.1888±0.045）、（0.4471±0.063）、（0.1834±0.039）、（0.3224±0.044）、（0.1788±0.042）,VTA中GFAP免疫反应的强度分别为（0.1538±0.045）、（0.2768±0.056）、（0.1808±0.036）、（0.2512±0.051）、（0.1427±0.034）.吗啡成瘾大鼠VTA中TH和GFAP免疫反应的强度持续高于淀粉组（P＜0.05）,安君宁治疗12d或30d能显著性降低VTA中TH、GFAP免疫反应强度（P＜0.05）,逐渐接近淀粉对照组.结论 吗啡成瘾能导致大鼠VTA内的一些可塑性的损伤,而安君宁能促进这些损伤的修复,提示其可用于阿片类成瘾的治疗.

Objective To explore the effects of ANjUNNING on tyrosine hydroxylase（TH） and glial fibrillary acidic protein（GFAP） in ventral tegmental area（VTA） of morphine dependent SD rats. Methods Thirty male Sprague-Dawley rats weighing 180～220g were assigned randomly into five groups： Normal group, treated with saline; P12E group, abstinent after the establishment of morphine dependence mode and treated with ANJUNNING for 12 days;P12EC group, abstinent after morphine dependence and treated with saline for 12 days; P30E group, abstinent from morphine dependence and treated with ANJUNNING for 30 days; P30EC group, abstinent from morphine dependence and treated with saline for 30 days, The morphine dependence mode was established by injecting intrapertoneally morphine in increasing dose （Smg/kg ～ 50mg/kg, twice a day） for 10 days. Administration of ANJUNNING in dose of 3g/kg was twice a day and from the termination of morphine injection, accordingly starch in the same volume to ANJUNNING was administrated to interventlon-controlled group at the same style. After the rats were anesthetized and infused, the brain were removed and cut into sections. The levels of the protein of TH, GFAP in VTA determined by immunohistochemistry. Results The levels of TH, GFAP in VTA of five groups （ Normal ,P12EC ,P12E, P3OEC, P30E） are 0. 1888 ±0.045,0. 4471±0. 063,0. 1834± 0. 039,0.3224 ± 0. 044,0. 1788 ± 0. 042, The levels of GFAP in VTA of five groups are 0. 1538 ± 0. 045,0.2768 ± 0. 056,0. 1808±0. 036,0.2512 ± 0. 051,0. 1427 ± 0. 034. Compared with intervention-controlled group, ANJUNNING markedly inhibited the expression of TH, GFAP in VTA. （ P 〈 0.05 ） ; Conclusion The functional disturbance of dopamine implicated in the craving and drug-seeking behavior mechanisms, ANJUNNING may promote recovery of the functional disturbance of the systems. It may be the mechanism by which ANJUNNING alleviated the heroin protracted abstinence symptoms, especially craving, in clinical application.