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目的了解CdCl2对腺垂体的损伤以及细胞凋亡发生与p38MAPK、ERK12表达的关系,为了解镉致腺垂体毒作用的分子机制提供科学依据。方法采用健康雄性SD大鼠进行整体试验(n=12),分别每天经口灌胃给予0、1.0、2.0、4.0mgkgbwCdCl2,6周后取腺垂体进行指标检测;离体试验采用酶解分离大鼠之原代腺垂体细胞,分别以0、1.56、3.12、6.25、12.50、25.00、50.00、100.00μmolLCdCl2处理,收获细胞进行检测;特异性阻断剂阻断凋亡效应的试验采用2.65μmolLp38MAPK的特异阻断剂SB203580或10μmolLERK12激酶的特异阻断剂U0126处理细胞,然后以3.12μmolL或100.00μmolL的CdCl2处理细胞后进行相应的检测。检测指标包括:TUNEL法和流式细胞术等检测凋亡。结果整体实验和离体实验结果均显示CdCl2以剂量依赖方式诱导腺垂体细胞发生凋亡(P<0.05);特异性阻断剂效应的研究结果显示2.65μmolLSB203580和10μmolLU0126对TUNEL阳性细胞的相对灰度和凋亡细胞率均有一定的影响。结论在一定的剂量条件下,CdCl2影响腺垂体激素分泌水平,并导致发腺垂体细胞凋亡,MAPKs家族成员p38MAPK和ERK12激酶通路在凋亡发生过程中可能发挥一定的作用。
Objective To investigate the relationship between the damage of pituitary gland and the expression of p38 MAPK and ERK12 in cadmium chloride (CdCl2), and to provide a scientific basis for understanding the molecular mechanism of cadmium-induced adenohypophysis. Methods Whole body test was conducted in healthy male SD rats (n = 12). The rats were orally gavaged with 0,1.0,2.0,4.0mgkgbwCdCl2 orally for 6 weeks respectively. Rat primary adenohypophyseal cells were treated with 0, 1.56, 3.12, 6.25, 12.50, 25.00, 50.00 and 100.00 micromol L CdCl 2, respectively. Cells were harvested for detection. The blocking effect of specific blockers was detected by using 2.65 μmol Lp38MAPK Cells were treated with inhibitor U206, a blocker SB203580 or 10 umol LERK12 kinase, and then treated with 3.12 umol L or 100 umol l of CdCl2 for the corresponding detection. Detection indicators include: TUNEL assay and flow cytometry and other apoptosis. Results The results of the whole experiment and in vitro experiments showed that CdCl2 induced apoptosis of adenohypophyseal cells in a dose-dependent manner (P <0.05). The results of specific blockade showed that the relative gray value of 2.65μmol LSB203580 and 10μmolLU0126 to TUNEL positive cells And the rate of apoptotic cells have a certain impact. Conclusions CdCl2 affects the secretion of pituitary gland hormones and induces the apoptosis of pituitary cells in a dose-dependent manner. The p38MAPK and ERK12 kinase pathways of MAPKs may play a role in the process of apoptosis.