Pitx3在帕金森模型大鼠黑質(zhì)區(qū)表達(dá)情況的研究

徐 海

[摘要] 目的 研究帕金森病(Parkinsons disease ,PD) 大鼠黑質(zhì)區(qū)pitx3 基因的表達(dá)情況。方法將6-OHDA注入紋狀體內(nèi)制作PD大鼠模型,于不同時(shí)間點(diǎn)處死,觀察pitx3基因的表達(dá)情況。結(jié)果同正常組、假手術(shù)組比較,PD模型大鼠損傷側(cè)黑質(zhì)區(qū)的pitx3基因在模型成功后第一周表達(dá)明顯下降,15天后少量增加,30天后也未恢復(fù)到正常水平。結(jié)論6-OHDA紋狀體內(nèi)注射制作的PD大鼠模型中腦黑質(zhì)區(qū)pitx3的表達(dá)明顯下降與DA能神經(jīng)元的死亡直接相關(guān),2周后pitx3表達(dá)增加可能與自體神經(jīng)干細(xì)胞的增殖與分化有關(guān)。

[關(guān)鍵詞] 帕金森病; 黑質(zhì)區(qū);pitx3

[中圖分類號(hào)] R742 [文獻(xiàn)標(biāo)識(shí)碼] A[文章編號(hào)] 1004-8650(2009)06-019-02

The gene expression of Pitx3 in substantia nigra of rat model of Parkinsons disease

XU Hai

[Abstract] Objective To explore the gene expression of Pitx3 in substantia nigra of rat model of Parkinsons disease. Methods Rat model of Parkinsons disease was made by injecting the 6-hydroxydopaminin(6-OHDA)in the striatum. The rats were killed at different time points, and their brains were taken out. Using the methods of immunofluorescence, we deteded the gene expression of Pitx3 in substantia nigra of hemi-parkinsonian rats. Results Campared with the controls, the number of pitx3-positive cells was significantly decreased in the first week after PD models were set up, however, there was also a slight increase of pitx3-positive cells 15 days after and no further increase 30 days after. Conclusion The decreasing expression of Pitx3 in substantia nigra of PD rat model is probably related to the apoptosis of the dopaminergic neurons. Furthermore, the augmentation of pitx3 gene expression 15 days after may be associated with the proliferation and differentiation of neural stem cells in PD rat models.

[Key words] parkinsons disease ; substantia; pitx3

帕金森病(Parkinsons disease,PD)是常見的神經(jīng)系統(tǒng)的退行性疾病,以震顫、肌強(qiáng)直及運(yùn)動(dòng)遲緩為主要臨床表現(xiàn),發(fā)病機(jī)制與腦內(nèi)的黑質(zhì)-紋狀體系統(tǒng)多巴胺含量減少有關(guān)。研究PD模型大鼠黑質(zhì)區(qū)多巴胺能神經(jīng)元的死亡與再生具有重要的理論價(jià)值和臨床意義。pitx3基因?yàn)榇贵w同源盒基因家族的成員之一,其表達(dá)產(chǎn)物pitx3[1-2]是決定中腦黑質(zhì)多巴胺(dopamine,DA)能神經(jīng)元表型、終末分化和生存維持的關(guān)鍵性特異性轉(zhuǎn)錄因子。因此,本實(shí)驗(yàn)通過(guò)6-OHDA紋狀體內(nèi)注射制作PD大鼠模型,觀察其腦內(nèi)黑質(zhì)區(qū)pitx3基因的表達(dá)情況。

1材料與方法

1.1PD模型的制作

雄性健康SD大鼠,體重200-250g,反復(fù)檢測(cè)確認(rèn)其無(wú)旋轉(zhuǎn)行為。0.4%的戊巴比妥鈉(40mg/kg)腹腔注射麻醉,固定于立體定位儀上,常規(guī)消毒,開顱。立體定位坐標(biāo)為兩點(diǎn):(參照Paxinos, Waston 大鼠圖譜),第一點(diǎn)AP1 0.2,ML1 3,DV1 5.5用微量注射器緩慢注入6-OHDA(濃度3μg/μl)3μl,留針10分鐘。提針至DV 4.5,進(jìn)藥3μl,第二點(diǎn)AP20.7,ML2 3DV25.5,進(jìn)藥過(guò)程同前。對(duì)照組定位點(diǎn)同前,注入等量的生理鹽水代替6-OHDA。……