论文部分内容阅读
目的:观察重组人核糖核酸酶抑制因子对小鼠脑内多巴胺能神经元保护作用。方法:将重组的pGEX-6p-1-hRI转化到大肠杆菌Rosetta表达菌中,并对产物提纯得到谷胱甘肽核糖核酸酶抑制因子(GST-RI),选取健康C57BL/6品系小鼠,随机分为5组:模型对照组皮下注射MPTP(溶于三蒸水,pH8.5)20mg/kg,连续8d;实验给药组(低剂量组腹腔注射GST-RI0.5mg/kg);高剂量组(腹腔注射GST-RI1.0mg/kg);空白给药组(腹腔注射GST1.0mg/kg)每日分别2次注射,连续13d,同时予MPTP造模;空白对照组给等量生理盐水。各组同时进行爬杆、悬挂行为学实验。实验结果后,动物断头取脑,取纹状体,应用高效液相色谱法(HPLC)测定小鼠左侧纹状体内多巴胺(DA)和高香草酸(HVA)的含量;免疫组织化学方法检测黑质酪氨酸羟化酶(TH)的免疫反应活性。结果:PD模型小鼠均出现不同程度的行为障碍,得分较空白对照组明显降低;而实验给药组小鼠得分则显著回升,以高剂量组尤为明显。TH免疫组化结果显示:与空白对照组比较,PD模型小鼠中脑黑质致密部TH免疫反应阳性神经元明显减少;实验组TH阳性神经元数目明显多于对照组.HPLC结果显示:PD模型小鼠较空白对照组明显降低,HVA/DA比值升高,给药组小鼠纹状体内DA和HVA含量明显恢复,高剂量组HVA/DA比值降至正常。结论:GST-RI对PD小鼠黑质DA能神经元的形态与功能具有明显的保护作用。
Objective: To observe the protective effect of recombinant human ribonuclease inhibitor on dopaminergic neurons in mice brain. Methods: Recombinant pGEX-6p-1-hRI was transformed into E.coli Rosetta. The product was purified to obtain glutathione S-transferase inhibitor (GST-RI). Healthy C57BL / 6 mice were selected, The rats in the model group were randomly divided into 5 groups: MPTP (20 mg / kg in distilled water, pH8.5) injected subcutaneously in the model control group for 8 days; experimental group (GST-RI 0.5 mg / kg) Dose group (GST-RI1.0mg / kg); blank control group (intraperitoneal injection of GST1.0mg / kg) were injected twice daily for 13 days, meanwhile MPTP was used for modeling. In blank control group, brine. Each group at the same time climbing rod, hanging behavior test. After the experimental results, the animals were decapitated and the striatum was taken. The contents of dopamine (DA) and homovanillic acid (HVA) in the left striatum were determined by high performance liquid chromatography (HPLC). The immunohistochemistry Immunoreactivity of nigrosine tyrosine hydroxylase (TH) was measured. Results: PD model mice all showed different degrees of behavioral disturbance, the score was significantly lower than the blank control group; while the experimental administration group mice score was significantly increased, especially in the high-dose group. TH immunohistochemistry results showed that compared with the blank control group, the number of TH immunoreactive positive neurons in substantia nigra compact zone in PD model mice was significantly decreased; the number of TH positive neurons in the experimental group was significantly more than that in the control group.HPLC results showed that PD Compared with the blank control group, the model mice decreased significantly and the HVA / DA ratio increased. The contents of DA and HVA in the striatum of the mice in the treatment group were significantly restored, while the HVA / DA ratio in the high dose group was reduced to normal. Conclusion: GST-RI has a significant protective effect on the morphology and function of dopaminergic neurons in substantia nigra of PD mice.