摘要
目的研究继发性甲状旁腺功能亢进症大鼠在病程中Gcm2的表达变化以及与甲状旁腺细胞增殖变化之间的关系。方法5/6肾切除结合高磷饲养建立大鼠继发性甲状旁腺功能亢进症模型,第1、4、7、14、21、28天取甲状旁腺和血清标本,ELISA检测血清i-PTH浓度变化,Real-time PCR和Western blot检测Gcm2mRNA和蛋白表达的变化,免疫组化观察不同时间点甲状旁腺细胞的增殖变化情况。结果与假手术对照组比较,造模后模型组i-PTH显著升高(P<0.01);模型组Gcm2mRNA的表达随时间进展升高趋势逐渐明显,在第1、4、7天仅缓慢升高,无统计学差异(P>0.05),第14天始升高趋势明显(P<0.01),于第21天升高至最高点,为假手术对照组的9.92倍(P<0.01),第28天表达量稍有下降(P<0.01);模型组Gcm2的蛋白表达呈先下调后升高的趋势,第1天先有所下调,第4天进行性升高,第7天稍有波动,第14天后一直处于显著升高的表达状态,于第21天到达最高点,第28天稍有回落(P<0.01);模型组甲状旁腺细胞增殖在第1天便显著增强(P<0.01),第4天达最高(P<0.01),之后开始回落,至第14天回落到与假手术对照组相近的水平,一直到实验结束。结论在继发性甲状旁腺功能亢进症发病过程中,Gcm2的表达变化和甲状旁腺细胞增殖在时间进程上有相反的趋势,两者的交互作用可能是导致甲状旁腺功能亢进的部分原因。
Objective To determine the expression profile of glia cells missing 2 （Gcm2） in rats during the course of secondary hyperparathyroidism and its relationship with parathyroid cell proliferation. Methods A total of 42 female SD rats, at an age of 6 to 8 weeks, weighting 180 to 220 g, were randomly divided into sham-operation group （n=6） and hyperparathyroidism group （n=36）. Rat model of secondary hyperparathyroidism was established by 5/6 nephrectomy combined with following high phosphorus feeding. Parathyroid glands and serum were collected after 6 rats of the model group were killed at the following intervals of 1, 4, 7, 14, 21 and 28 d, respectively. Serum level of intact parathyroid hormone （i-PTH） was detected by ELISA. The expression of Gcm2 at mRNA and protein levels was observed by real-time PCR and Western blotting, respectively. Parathyroid cell proliferation were observed by immunohistochemical assay. Results Compared with the sham-operation group, i-PTH levels were significantly elevated at all time points in the model group （P〈0.01）. In the model group, the mRNA expression of Gcm2 was increased gradually with time elapse, though without significant difference in the days 1, 4 and 7, but significantly higher in the day 14 （P〈0.01）, and reached the highest level in the day 21, which were increased by 9.92 fold when compared with sham-operation group （P〈0.01）, and then the level was decreased mildly in the day 28 （P〈0.01）. While, the expression of Gcm2 protein was slightly weakened in the first day, then gradually elevated in the day 4, and continued to raise till the highest level in the day 21 except a little fluctuation in the day 7, but the level was also decreased in the day 28 （P〈0.01）. The proliferative rate of parathyroid cells was very high in the day 1 （P〈0.01）, and reached the peak in the day 4 （P〈0.01）, and then became declined till to the normal rate since the day 14. Conclusion In the pathogenesis of secondary hyperparathyroidism, Gcm2 expression and parathyroid cell proliferation showed an opposite trend. The interaction between them may partly contribute to the hyperfunction of the parathyroid.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第11期1080-1083,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(81270870)~~
