法舒地尔抗氧化应激保护LPS-PD小鼠黑质多巴胺神经元作用的研究  

Rho Kinase Inhibitor Fasudil Protects Dopamine Neurons of Substantia Nigra through Alleviating Oxidative Stress in LPS-PD Mouse

作　　者：赵永飞 姚恺 张梦奇 肖保国 ZHAO Yong-fei;YAO Kai;ZHANG Meng-qi;XIAO Bao-guo(Department of Neurology,Jinshan Hospital Affiliated to Fudan University,Shanghai 201508,China;Department of Neurology,Huashan Hospital Affiliated to Fudan University,Shanghai 200040,China;Institute of Neurology,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属金山医院神经内科,上海201508 [2]复旦大学附属华山医院神经内科,上海200040 [3]复旦大学神经病学研究所,上海200040

出　　处：《中国临床神经科学》2020年第5期481-487,共7页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金项目(编号:81371414)。

摘　　要：目的观察Rho激酶抑制剂法舒地尔(Fa)对脂多糖(LPS) PD模型小鼠中脑黑质多巴胺(DA)神经元的保护作用。方法采用C57BL/6小鼠两侧鼻孔滴入LPS 20μL (1μg.μL-1),隔天1次,×60 d,制备小鼠LPS-PD模型。LPS-PD模型鼠随机分为LPS-PD+Fa治疗组(n=8,LPS滴鼻45 d起腹腔注射Fa 20 mg.kg-1.d-1,每天2次,×14 d)、LPS-PD组(n=8)和正常对照组(n=8,腹腔注射等量0.9%氯化钠注射液)。采用胶布黏附移除实验评估小鼠行为学变化,Western blot测定小鼠脑组织内酪氨酸羟化酶(TH)、Rho激酶2(ROCK2)、诱导型一氧化氮合酶(iNOS)、尼克酰胺腺嘌呤二核苷酸磷酸氧化酶(gp91phox)、血红素加氧酶(HO)-1表达,免疫荧光技术测定小鼠黑质纹状体区TH表达。分析Fa治疗后与氧化应激反应因子表达以及黑质DA神经元丢失之间的关系。结果与正常对照组比较,LPS-PD组黑质TH表达显著降低,ROCK2表达显著增高,iNOS和gp91phox表达显著增高(均P<0.05),而HO-1表达下降(P<0.05)。与LPSPD组比较,LPS-PD+Fa治疗组黑质TH表达显著增加,ROCK2、iNOS、gp91phox表达显著下降,HO-1表达增加(均P<0.05)。LPS-PD组运动功能显著下降;LPS-PD+Fa治疗组运动评分显著改善,与TH表达呈平行关系。结论 Fa降低ROCK2表达可以抑制氧化应激反应,保护并减少黑质DA神经元丢失,改善LPS-PD模型小鼠运动协调功能。Aim To survey the neuroprotective effects of Fasudil(Fa) against the dopamine(DA) neurons in the substantia nigra in the model of Parkinson’s disease(PD) induced with lipopolysaccharide(LPS). Methods PD model were induced with a bilateral intranasal instillation of 20 μL(1 μg.μL-1) LPS every other day for 60 days. C57 BL/6 mice were randomly divided into a LPS-PD group(n=8), a LPS-PD + Fa group(n=8, Fa was delivered intraperitoneally at 20 mg.kg-1 body weight as twice daily for fourteen consecutive days started on the 45 th day after the first LPS intranasal instillation), a control group(n=8, intraperitoneal injection of the same amount of 0.9% sodium chloride injection). Behavior activity was evaluated by adhesive tape removal test. The expression of TH, ROCK2, iNOS, HO-1, gp91 phox were detected by Western blot and immunohistofluorescence. The relationship between oxidative stress response factor expression and substantia nigra DA neuron loss after Fa treatment was analyzed. Results TH expression in the substantium of LPS-PD group was significantly decreased, ROCK2 expression was significantly increased, and iNOS and gp91 phox expression were significantly increased(both P<0.05), while HO-1 expression decreased(P<0.05) compared with the normal control group. The expression of TH, HO-1 was significantly improved, while the expression of ROCK2, iNOS, gp91 phox was decreased dramatically in the LPS-PD + Fa group compared to the LPS-PD group(both P<0.05). The motor scores of the LPS-PD group were significantly prolonged, and those of the LPS-PD+Fa treatment group were significantly improved, which was parallel to TH expression. Conclusion The rescue of DA neurons in substantia nigra along with improvement of movement coordination in LPS-PD model may be benefited partly from the inhibition of oxidative stress treated with Rho kinase inhibitor Fa.

关 键 词：帕金森病 酪氨酸羟化酶 多巴胺 RHO激酶 氧化应激 

分 类 号：R742.5[医药卫生—神经病学与精神病学] Q198[医药卫生—临床医学]