【摘要】 背景:D-半乳糖衰老模型具有与人类自然衰老相类似的生化改变,蛋白磷酸酯酶1在D-半乳糖脑老化小鼠表达降低是否与脑老化过程有关还有待于进一步研究。目的:探讨APP17肽及救脑益智胶囊水提液对D-半乳糖脑老化小鼠海马蛋白磷酸酯酶1表达的影响。设计:随机对照观察。单位:首都医科大学基础医学院病理学教研室和宣武医院脑老化研究室。材料:实验于2003-07/2004-07在首都医科大学实验动物中心完成,选用雄性昆明小鼠40只。购自中国医学科学院药物研究所。方法:将昆明小鼠随机分为5组:正常对照组、D-半乳糖对照组、APP17肽治疗组、中药小剂量治疗组、中药大剂量治疗组,每组动物8只。脑老化小鼠模型D-半乳糖组,APP17肽治疗组,中药小剂量治疗组,中药大剂量治疗组4组每日皮下注射半乳糖50mg/kg体质量,同时在APP17肽治疗组皮下注射APP17肽,每次每只小鼠注射含APP17肽0.35μg的生理盐水0.1mL,1次/d,连续注射3个月。中药治疗组分别给予0.3g/kg和1.0g/kg的救脑益智水提液(由北京朝阳门医院和山西曲沃传统医学研究所研制,其主要成分有:党参、白术、龟甲、穿山甲等)灌胃,1次/d。两个对照组每天注射和灌注等量的生理盐水,3个月后取脑组织做蛋白磷酸酯酶1免疫组织化学染色。主要观察指标:免疫组织化学染色检测各组动物海马蛋白磷酸酯酶1的表达。结果:40只小鼠均进入结果分析,无脱失值。D-半乳糖小鼠和大剂量救脑益智胶囊水提液治疗组小鼠海马蛋白磷酸酯酶1阳性细胞数目少,染色淡;正常小鼠、APP17肽保护和救脑益智胶囊水提液小剂量治疗的D-半乳糖小鼠海马阳性反应神经元数目多,胞浆与突起深染。结论:抑制凋亡的蛋白磷酸酯酶1在D-半乳糖小鼠海马表达降低。APP17肽和小剂量救脑益智胶囊水提液治疗能影响D-半乳糖脑病小鼠脑内蛋白磷酸酯酶1的表达,使之接近正常。而大剂量救脑益智胶囊水提液治疗无此作用,其量效关系特点还需要进一步考察验证。更多还原

【Abstract】 BACKGROUND: D-galactose-induced aging animal model is similar to human natural aging. Whether the expression of protein phosphatase-1 (PP-1) in the brain of D-galactose-induced aging mice is related to cerebral aging process or not should be researched further. OBJECTIVE: To investigate the effect of the APP17 peptide and the liquid extract of jiunao yizhi capsule on regulating the expression of PP-1 in hippocampal neurons of the aging mice induced by the D-galactose (D-gal). DESIGN: A random controlled study. SETTING: Department of Pathology, College of Basic Medical Sciences, Capital University of Medical Sciences and Beijing Research Laboratory for Brain Aging, Xuanwu Hospital of Capital University of Medical Sciences. MATERIALS: The study was completed between July 2003 and July 2004 in the Experimental Center of Capital University of Medical Sciences. Forty male Kunming mice (SPF grade) with a body mass from 28 g to 32 g were purchased from Chinese Academy of Medical Sciences Institute of Material Medical. METHODS: Kunming mice were randomly divided into 5 groups: control group, D-gal group, APP17 peptide treatment group, low dose herb treatment group, and high dose herb treatment group with 8 mice in each group. In D-gal group, APP17 peptide treatment group, low dose herb treatment group and high dose herb treatment group, galactose was injected subcutaneously (50 mg/kg). Meanwhile, 0.1 mL normal saline containing 0.35 μg of APP17 peptide was injected subcutaneously into the mice in APP17 peptide treatment group, once a day for 3 months; liquid extract of jiunao yizhi capsule (provided by Beijing Chaoyangmen Hospital and Shanxi Quwo Traditional Medical Institute; the main component: dangshen, baizhu, guijia and chuanshanjia, etc.) was perfused by stomach (0.3 g/kg and 1.0 g/kg respectively) in low dose herb treatment group and high dose herb treatment group, once a day. And equivalent normal saline was injected and perfused in the two control groups. After 3 months of survival, the mice were killed and their brains were cut into sections. The immunohistochemical staining of these sections was then performed with PP-1 antibody. MAIN OUTCOME MEASURES: The results of immunohistochemical staining analysis of PP-1. RESULTS: Forty mice entered the final analysis without any loss. PP-1 positive cells in the hippocampus were poorly stained in the D-gal mice. In contrast, PP-1 positive neurons were widely distributed in the hippocampus of those normal mice, the APP17 peptide-treated D-gal mice and the high liquid extract of raw herb-treated D-gal mice. These cells were darkly stained in cytoplasm. The unexpected result was that in the low liquid extract of raw herb-treated D-gal mice the number of PP-1 positive neurons did not increase to normal. CONCLUSION: The results demonstrated that the expression of PP-1 decreased in the hippocampus of D-gal mice. The APP17 and low dose liquid extract of raw herbs can regulate the distribution of PP-1 in the brain of D-gal mice and make them recover to normal situation.更多还原