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目的探讨抗帕颗粒对1-甲基-4-苯基~(-1),2,3,6-四氢吡啶(MPTP)帕金森病(PD)模型小鼠黑质纹状体酪氨酸羟化酶(TH)阳性神经元的影响。方法 90只健康雄性C57BL/6小鼠,鼠龄8~12周,并随机分为3组,即正常对照组30只、PD模型对照组30只、PD模型干预组30只;MPTP腹腔注射(40 mg·kg~(-1)·d~(-1)×7 d)制备小鼠PD模型;正常对照组及PD模型对照组予生理盐水1 mL·d~(-1)灌胃,PD模型干预组给予抗帕颗粒40 mg·kg~(-1)·d~(-1)灌胃,连续喂养4个月;黑质纹状体切片、HE染色、免疫组织化学染色TH神经元及Western blotting检测TH蛋白的表达量。结果 (1)正常对照组30只(30/30只)最终均存活,PD模型对照组4个月存活27只(27/30只),PD模型干预组4个月存活28只(28/30只);(2)PD模型对照组、PD模型干预组小鼠每次注射MPTP后先有短暂兴奋[持续(7.61±2.17)min],表现为四处窜跳,随即出现全身中重度震颤,皮毛及尾巴时有竖立,活动减少,持续(24.23±3.89)min后震颤消失,随后出现活动减少;(3)HE染色显示正常对照组大量褐色TH阳性细胞,PD模型对照组TH阳性细胞数明显减少,PD模型干预组TH阳性细胞数有所增加;(4)免疫组织化学染色后经Imagepro-Plus 5.1系统分析,正常对照组TH阳性细胞面积为64 145μm~2,高倍镜下可见大量胞质为褐色颗粒的TH阳性细胞;PD模型对照组TH阳性细胞染色面积为40 012μm2,高倍镜下见TH细胞数明显减少;PD模型干预组TH阳性细胞染色面积为60 952μm~2,高倍镜下见TH阳性细胞数较PD模型对照组增加;(5)Western blotting检测显示正常对照组TH蛋白表达量与PD模型对照组和PD模型干预组比较均有明显差异(P<0.001),PD模型对照组TH蛋白表达量与PD模型干预组比较也有明显差异(P<0.05)。结论抗帕颗粒可使PD小鼠黑质纹状体中多巴胺能神经元一定程度地减少丢失,对多巴胺能神经元的数量、形态及功能具有一定的保护作用。
Objective To investigate the effects of Kangpai Granules on nigrostriatal tyrosine in mice with Parkinson’s disease (PD) of 1-methyl-4-phenyl- (-1), 2,3,6-tetrahydropyridine Hydroxylase (TH) positive neurons. Methods Ninety healthy male C57BL / 6 mice aged 8-12 weeks were randomly divided into three groups: 30 in normal control group, 30 in PD model control group and 30 in PD model intervention group. MPTP was injected intraperitoneally 40 mg · kg -1 · d -1 × 7 d). The normal control group and PD model control group were treated with normal saline (1 mL · d -1) The model intervention group was given intragastric administration of 40 mg · kg -1 d -1 of Baopai Granule and fed continuously for 4 months. The nigrostriatal sections were stained with HE and immunohistochemical staining for TH neurons and Western blotting was used to detect the expression of TH protein. Results (1) Thirty (30/30) rats in the normal control group survived in the end, while the control group of the PD model survived 27 months (27/30) in the 4 months and the 28 rats in the PD model intervention group survived 4 months Only); (2) PD model control group, PD model intervention group mice were transiently excited after each injection of MPTP [continuous (7.61 ± 2.17) min], manifested as jumping around, then appeared in moderate and severe tremors, fur (3) HE staining showed a large number of brown TH positive cells in the normal control group, while the number of TH positive cells in the PD model control group was significantly decreased (P <0.01) , PD model intervention group increased the number of TH positive cells; (4) Immunohistochemical staining by Imagepro-Plus 5.1 system analysis, the normal control group TH positive cell area of 64 145μm ~ 2, a large number of cytoplasm can be seen under high magnification The number of TH cells in the PD model control group was 40 012μm2, and the number of TH cells was significantly reduced in the high power microscope. The staining area of TH positive cells in the PD model intervention group was 60 952μm2, The number of positive cells increased compared with the PD model control group. (5) Western blotting showed that the normal control The expression of TH protein in PD model group was significantly lower than that in PD model group and PD model group (P <0.001). There was also a significant difference (P <0.05) between TH group and PD model group. CONCLUSION: PPA can reduce the loss of dopaminergic neurons in substantia nigra and striatum of PD mice to a certain extent, and protect the number, morphology and function of dopaminergic neurons.