重复注射6-羟多巴胺建立帕金森病动物模型  

Establishment of the animal model of Parkinson disease by repeated injection of 6-hydroxydopamine

作　　者：邓汝东[1] 李伊为[1] 汪华侨[2] 陈东风[1] 张赛霞[1] 赵丹[1] 孟令杰[1] 周健洪[1] 黎晖[1] 

机构地区：[1]广州中医药大学基础医学院,广东广州510006 [2]中山大学中山医学院,广州510080

出　　处：《解剖学研究》2007年第6期401-404,共4页Anatomy Research

基　　金：国家自然科学基金资助项目(30472272;30772861)

摘　　要：目的探讨通过大鼠中脑内重复注射 6-羟多巴胺 (6-hydroxydopamine,6-OHDA)建立高效、稳定、可靠的帕金森病动物模型。方法将 60 只雄性 SD 大鼠随机分为一次打击组和二次打击组,经腹腔注射 4%水合氯醛(40 mg / 100g)麻醉,脑立体定位仪固定,于左侧黑质致密部(SNC)和中脑腹侧被盖区(VTA)分别注射 6-羟多巴胺 (6-OHDA,2 g / L,2μl),二次打击组一周后在相同位置重复注射同剂量的 6-OHDA,建立帕金森模型,观察其行为改变,通过 HE、TH 和 DIG-dUTP 染色观察其细胞形态的改变及凋亡情况。结果经过二次打击的大鼠,模型成功率(旋转周数 >7 r / min)为 86.7%,明显多于一次打击组的 33.3%;HE 染色显示,二次打击组凋亡细胞的阳性率(37.12%)明显多于一次打击组(21.25%);DIG-dUTP 染色显示,一次打击组大鼠左侧中脑黑质区神经细胞肿胀的数量多 ,凋亡数量少,凋亡细胞阳性率为 20.73%,二次打击组细胞凋亡数显著增多,凋亡细胞阳性率达 36.03%;TH 染色显示,二次打击组 TH 阳性细胞数明显减少,TH 细胞阳性率(18.61%)显著低于一次打击组(36.55%)。结论通过二次打击建立帕金森病动物模型成功率高于一次打击。Objective To establish an effective, stable and reliable model of Parkinson disease by repeated injection of 6-hydroxydopamine （6-OHDA）. Methods The 60 male Spague-Dawley （SD） rats of clean grade （weighing from 200 to 210 g） were randomly divided into two groups： one-hit group, two-hit group. One hit： the rats were anesthetized by intraperitoneal injection of 4% Chloral Hydrate （40 mg/100 g）, and fixed on the stereotaxis instrument. 6-hydroxy dopamine （2 g/L, 2 μl） was injected into the left substantia nigra pars compacta （SNC） and ventral tegmental area （VTA） in the midbrain to develop the Parkinson disease rat model. Two hit： one week after the first hitr 6-hydroxydopamine with the same dosage was injected in the same position to develop the Parkinson disease model. Then the behavior changes of rats were observed, and morphological changes and the extent of apoptosis were assayed by Hematoxylin ＆ Eosin （HE） staining, Tyrosin hydroxylase （TH） staining and digoxigenin-11-dUTP （DIG-dUTP） staining. Results The success rate of model in the rats suffering two-hit was 86.7% （rotation rate exceeded 7 r/min） which was significantly more than 33.3% in the rats suffering one-hit. HE staining showed that the positive apoptotic cells in two-hit group （37.12%） were much more than that in one-hit group （21.25%）. DIG-dUTP staining showed that in one-hit group, there were many swollen nerve cells and less apoptosis nerve cells in the left substantia nigra of midbrain of rats, and the positive apoptotic cells represented 20.73%; in two-hit group,the apoptosis nerve cells increased dramatically, and the positive apoptotic cells accounted for 15.3%. TH staining indicated that the TH positive cells in two-hit group（18.61%） decreased significantly, and were dramatically less than TH positive cells in one-hit group （36.55%）. Conclusion The success rate of establishing Parkinson disease model by two-hit was higher than that by one-hit.

关 键 词：帕金森病 6-羟多巴胺 动物模型 黑质 

分 类 号：R-332[医药卫生]