黄芪对N-甲基-N-亚硝脲诱导大鼠光感受器细胞凋亡的预防途径(英文)

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背景:视网膜色素变性是非炎症性、双侧进行性的视网膜变性,其特征是光感受器细胞因发生凋亡而丢失,最终导致失明。中药黄芪在阻止该病的进程上显示出了良好的前景。目的:观察黄芪对N-甲基-N-亚硝脲致SD大鼠视网膜损伤的保护作用。设计:随机对照动物实验。单位:新乡医学院药学院。材料:实验于2004-03/12在中山大学中山眼科中心药理实验室完成。雌性SD大鼠114只,由中山大学中山医学院动物中心提供,N-甲基-N-亚硝脲为美国Sigma公司产品,黄芪注射液为成都地奥九泓制药厂生产(生产批号:国药准字Z99060535,2mL/支,主要成分为黄芪)。方法:选用雌性SD大鼠114只,30只用于视网膜层形态学分析;30只用于细胞凋亡检测;54只用于胞核内核因子κBp65活性的检测。采用抽签法随机分组,每组6只。于大鼠生后47d,分别经腹腔注射不同剂量的黄芪(2.5,5和10g/kg),1次/d,除对照组外,其余组的大鼠同时于生后50d腹腔注射N-甲基-N-亚硝脲60mg/kg。在N-甲基-N-亚硝脲处理不同时间后处死动物,取眼球,视网膜形态学分析测量周边视网膜的总厚度,TUNEL试剂盒检测光感受器细胞凋亡,转录因子试剂盒分析核因子κBp65的活性。主要观察指标:各组视网膜厚度、凋亡指数和胞核内核因子κBp65的活性比较。结果:黄芪可剂量依赖性地显著增加周边视网膜总厚度和降低N-甲基-N-亚硝脲引起的光感受器细胞凋亡指数。黄芪注射液10g/kg还可时间依赖性地上调视网膜细胞胞核内核因子κBp65的活性。但其对N-甲基-N-亚硝脲引起的中心视网膜损伤无明显的保护作用。结论:黄芪通过上调视网膜细胞胞核内核因子κBp65的活性,抑制光感受器细胞凋亡,从而部分地保护N-甲基-N-亚硝脲引起的视网膜损伤。 BACKGROUND: Retinitis pigmentosa is a non-inflammatory, bilateral progressive retinal degeneration, characterized by the loss of photoreceptor cells due to apoptosis, which ultimately leads to blindness. The Chinese medicine Astragalus has shown good prospects in preventing the disease. OBJECTIVE: To observe the protective effect of Astragalus membranaceus on retinal damage induced by N-methyl-N-nitrosourea in SD rats. Design: Randomized controlled animal experiments. Unit: School of Pharmacy, Xinxiang Medical College. MATERIALS: The experiment was performed at the Laboratory of Pharmacology, Zhongshan Eye Center, Sun Yat-sen University from March to December 2004. 114 female SD rats were provided by the Animal Center of Zhongshan Medical College of Sun Yat-sen University, N-methyl-N-nitrosourea was produced by Sigma, and Huangqi injection was produced by Chengdu Dio Jiuhao Pharmaceutical Factory (production batch number: Sinopharm. Quasi-Z99060535, 2mL/branch, mainly composed of Astragalus membranaceus. METHODS: A total of 114 female SD rats were selected and 30 were used for morphological analysis of the retina layer; 30 were used for apoptosis detection; 54 were used for the detection of nuclear factor κB p65 activity. Randomly grouped by lottery, 6 in each group. After rats were born 47 days later, different doses of jaundice (2.5, 5, and 10 g/kg) were injected intraperitoneally once a day. In addition to the control group, rats in the other groups were injected intraperitoneally with N-alpha for 50 days after birth. -N-nitrosourea 60 mg/kg. The animals were sacrificed after N-methyl-N-nitrosourea treatment for different periods of time. The eyeballs were taken and the total retinal thickness was measured by morphological analysis of the retina. The TUNEL kit was used to detect photoreceptor apoptosis and the transcription factor kit was used to analyze nuclear factor-κB p65. Activity. MAIN OUTCOME MEASURES: Comparison of retinal thickness, apoptotic index, and activity of nuclear factor kappa Bp65 in each group. RESULTS: Astragalus membranaceus significantly increased peripheral retinal total thickness and decreased photoreceptor apoptosis index induced by N-methyl-N-nitrosourea in a dose-dependent manner. Astragalus injection 10g/kg can also time-dependently upregulate the activity of nuclear factor kappa B p65 in retinal cells. However, it has no obvious protective effect on central retinal damage caused by N-methyl-N-nitrosourea. CONCLUSION: Astragalus membranaceus can partly protect the retinal damage caused by N-methyl-N-nitrosourea by up-regulating the activity of nuclear factor kappa B p65 in retinal cells and inhibiting photoreceptor cell apoptosis.
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