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目的 了解氯化镉诱导腺垂体细胞凋亡发生及半胱氨酸天冬氨酸特异性蛋白酶(procaspase- 9)mRNA表达的变化,为阐明镉致腺垂体毒作用的分子机制提供科学依据。方法 在整体实验中,将健康雄性SD大鼠 48只,分为 4组,每组 12只,分别每天经口灌胃给予 0、1 .0、2 .0、4. 0mg/kg氯化镉, 6周后进行腺垂体激素促肾上腺皮质激素、黄体生成素、卵泡刺激素水平、超微结构、原位末端标记(TUNEL)法检测细胞凋亡及procaspase 9mRNA表达检测;离体实验采用酶解分离大鼠之原代腺垂体细胞, 37℃、5%CO2 条件下培养 120h后分别以 0、1. 56、3 .12、6 .25、12.50、25 .00、50 .00、100 00μmol/L氯化镉处理 6h,收获细胞以TUNEL法检测细胞凋亡及原位杂交法检测procaspase- 9mRNA表达。结果 整体实验表明氯化镉作用后 3个剂量组血中促肾上腺皮质激素、黄体生成素和 1 0mg/kg氯化镉剂量组卵泡刺激素水平均降低,腺垂体细胞呈现明显的凋亡征象;整体和离体实验结果均显示氯化镉以剂量依赖方式诱导腺垂体细胞procaspase- 9mRNA表达的增强,且可使凋亡细胞增多,与对照组比较,差异均具有统计学意义。结论 在一定剂量条件下,氯化镉影响腺垂体激素分泌水平改变,并诱发腺垂体细胞凋亡,caspase -9通路在凋亡发生过程中可能发挥了一定的作?
Objective To investigate the apoptosis of adenohypophyseal cells and the expression of procaspase-9 mRNA induced by cadmium chloride (CdCl2) in order to provide a scientific basis for elucidating the molecular mechanism of cadmium-induced adenohypophysis. Methods In the whole experiment, 48 healthy male Sprague-Dawley rats were divided into 4 groups with 12 rats in each group. The rats were orally gavaged with 0,1. 0,2. 0,4. 0 mg / kg cadmium chloride After 6 weeks, the levels of adenocarcinoma, luteinizing hormone, follicle stimulating hormone, ultrastructure and TUNEL method were used to detect the apoptosis and procaspase 9 mRNA expression. In vitro experiments were carried out by enzymolysis Primary rat pituitary gland cells were isolated and cultured at 37 ℃ and 5% CO2 for 120 h. The cells were stained with 0, 1. 56, 3.12, 6.25, 12.50, 25.00, 50.00, 100.00 μmol / L cadmium chloride treatment 6h, cells were harvested to detect apoptosis by TUNEL method and in situ hybridization procaspase-9mRNA expression. Results The whole experiment showed that the serum levels of ACTH, luteinizing hormone and follicle-stimulating hormone in 10 mg / kg cadmium chloride dose group were decreased in 3 doses of cadmium chloride, and the apoptotic sign of adenohypophyseal cells was obvious. The results of whole and in vitro experiments showed that cadmium chloride induced the procaspase-9 mRNA expression in adenohypophyseal cells in a dose-dependent manner and increased the number of apoptotic cells. Compared with the control group, the differences were statistically significant. Conclusions Cadmium chloride can affect the secretion of pituitary gland hormones and induce the apoptosis of pituitary gland cells under certain dosage. The caspase -9 pathway may play a certain role in the process of apoptosis.