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目的探讨晚期糖基化终末产物受体(RAGE)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病(PD)模型小鼠脑中酪氨酸羟化酶(TH)表达量的影响。方法采用12周龄的C57BL/6雄性小鼠依据不同处理方法分为6组(均n=10):对照组;MPTP组;空载病毒阴性对照(RAGE-NC)组;目的基因阴性对照(siRNA-RAGE)组;RAGE-NC+MPTP组;siRNA-RAGE+MPTP组。携带空载siRNA的慢病毒(RAGE-NC)与携带抑制RAGE表达的目的 siRNA-RAGE慢病毒经脑立体定位仪定向注射于小鼠两侧黑质,根据分组情况给予腹腔注射MPTP 30mg·kg~(-1)或等量生理盐水(每周2次×5周)。5周后予小鼠断头取脑,利用免疫组织荧光染色法检测各组小鼠脑组织黑质中TH数量变化情况,采用Western blot法分别检测各组RAGE、caspase-3、TH蛋白表达水平。结果与RAGENC+MPTP组比较,siRNA-RAGE+MPTP组RAGE蛋白表达减少(0.782 8±0.139 6 vs 1.039 0±0.146 4,P<0.01),caspase-3蛋白表达减少(0.864 4±0.105 3 vs 1.240 0±0.080 7,P<0.001),TH蛋白表达量显著升高(1.114 0±0.201 1vs 0.771 1±0.211 3,P<0.05),差异有统计学意义。结论抑制RAGE表达可抑制凋亡反应,提高PD多巴胺能神经元模型中TH蛋白表达水平,有潜在的神经保护作用。
Objective To investigate the effects of advanced glycation end-products receptor (RAGE) on Parkinson’s disease (PD) -induced mice induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) Effect of tyrosine hydroxylase (TH) expression in brain. Methods C57BL / 6 male mice (12 weeks old) were divided into 6 groups (n = 10) according to different treatments: control group, MPTP group, RAGE-NC group, negative control group siRNA-RAGE) group; RAGE-NC + MPTP group; siRNA-RAGE + MPTP group. The lentivirus (RAGE-NC) carrying the empty siRNA and the target siRNA-RAGE lentivirus carrying the RAGE expression were targeted injected into the substantia nigra of both sides of the mice via the stereotaxic apparatus, and were given intraperitoneally MPTP 30 mg · kg ~ (-1) or normal saline (twice weekly x 5 weeks). After 5 weeks, the mice were decapitated and their TH content was measured by immunohistochemical staining. Western blot was used to detect the protein expression of RAGE, caspase-3 and TH in each group. Results Compared with RAGENC + MPTP group, the expression of RAGE protein in siRNA-RAGE + MPTP group was decreased (0.782 8 ± 0.139 6 vs 1.039 0 ± 0.146 4, P <0.01) and the expression of caspase-3 protein was decreased (0.864 4 ± 0.105 3 vs 1.240 0 ± 0.080 7, P <0.001), the expression of TH protein was significantly increased (1.114 0 ± 0.201 1vs 0.771 1 ± 0.211 3, P <0.05), the difference was statistically significant. Conclusion Inhibition of RAGE expression can inhibit the apoptotic response and improve the expression of TH protein in PD dopaminergic neuron model, which has the potential neuroprotective effect.