6-羟基多巴毁损间脑A11区域对缺铁饮食小鼠运动功能的影响

Effect of 6-OHDA lesioning diencephalic A11 nucleus on the locomotor acitivities of dietary iron deficiency mice

目的观察6-羟基多巴(6-hydroxydopamine,6-OHDA)定向毁损间脑A11核团对缺铁饮食小鼠运动功能的影响,探讨此方法建立不安腿综合征(Restless legs syndrome,RLS)模型的可行性。方法取60只4周龄C57BL/6雄性小鼠,30只予缺铁饮食(iron deficiency diet,ID),另外30只予正常饮食(normal diet,ND)。1个月后观察小鼠的运动活性,包括水平方向的运动次数(HACTV)、垂直方向的运动次数(VACTV)、总的运动距离(TOT-DIST)和运动时间(MOVTIME)。随后每组各取15只小鼠定向注射6-OHDA至双侧A11区域,其余小鼠注射PBS。并继续原来的饮食处理。毁损1个月后所有动物皮下注射多巴胺受体激动剂ropinirole,比较给药前后小鼠运动活性的变化。对A11区域进行酪氨酸羟化酶(tyrosine,TH)染色计数多巴胺(DA)神经元。结果饮食治疗1个月后,缺铁饮食组小鼠的三项运动活性均显著高于正常饮食组(均为P<0.01)。6-OHDA毁损A11核团可显著增加小鼠的各项运动活性(均为P<0.01)。D2受体激动剂ropinirole可以使小鼠增高的运动活性下降到基线水平。6-OHDA毁损可致A11区域DA能神经元明显减少。结论对C57BL/6雄性小鼠在缺铁饮食基础上进行A11区域的6-OHDA毁损建立的模型,能较好的模拟RLS的临床征象,可用作RLS的动物模型。

Objective To observe the effect of 6-hydroxydopamine （6-OHDA） lesioning dieneephalie All nucleus on the loccomotor acitivities of dietary iron deficiency mice and explore the probability of using the mouse fed with iron deficiency diet and lesioned with 6-OHDA as an animal model of restless legs syndrome （RLS）. Methods Half of sixty C57BL/6 male mice were given iron deficiency diet （ID） starting 28 days after birth, the other animals were given normal diet （ND）. Locomotor activities were measured after one month of dietary ID. Data were collected by computer included the number of horizontal locomotor activities （HACTV） and vertical locomotor activities （VACTV） , total distance traveled （TOTDIST） and move time （MOVTIME）. After that, half animals of each group were processed microinjection with 6-OHDA into bilateral Allnucleus. The rest mice were injected PBS. All the animals were continued previous dietary manipulation. After one month of All lesioned, dopamine receptor agonist ropinirole was administered intraperitoneally. The changes of locomotor activities were observed after thirty minutes of ropinirole administered. TH-positive neurons in All area were counted by immunohistochemistry. Results After one month of ID dietary treatment, the locomotor acitivities （ HACTV, TOTD1ST and MOVTIME） of the animals were significantly increased （P 〈 0. 001 for each comparison）. All the locomotor activities were also markedly increased after All lesioned （P 〈0. 01 for each comparison）. After ropinirole administered, all the increased measurements were decreased to baseline levels. TH-positive neurons in All region were significantly decreased after 6-OHDA lesions. Conclusions Both dietary ID and 6-OHDA lesions produce significant increase of the locomotor activities in all the animals. Dietary ID combined with A11 6-OHDA lesions in mice can mimic the clinical features of RLS, which could be used as an animal model of RLS.