P物质对大鼠垂体前叶细胞游离Ca^(2+)影响的研究

The Effect of Substance P on the Free Ca^(2+) Concentration of Cultured Anterior Pituitary Cells of the Rat

目的:探讨SP是否影响培养的大鼠AP细胞[Ca2+]i,在第二信使信息转导途径上进一步阐述SP产生生物学效应的机制。方法:原代培养成年雌性S-D大鼠的AP细胞48h,加入SP孵育不同时间,采用Fura-2/AM检测[Ca2+]i。结果:当SP浓度为100nmol/L,孵育时间由10min增加到30min时[Ca2+]i增加,但孵育时间为40min、50min时,[Ca2+]i呈现减少趋势,即最大反应的孵育时间为30min。当孵育时间为20min、30min、40min、50min时,[Ca2+]i分别为211±16nmol/L、369±51nmol/L、358±24nmol/L、239±36 nmol/L,与10min(117±17nmol/L)相比较,呈显著性差异(P<0.05)。结论:通过Fura-2/AM可精确反映[Ca2+]i,SP兴奋AP细胞SPR后可通过Ca2+来完成其部分的生物学效应,说明了SP对生殖轴(下丘脑-垂体前叶-卵巢/睾丸)有调控作用。

Objective：This thesis was to determine whether SP had an effect on the free Ca^2＋ concentration in primary cultured AP cells of the rat. We farther indicated that SP induced the action mechanism of physiology effect in the way of information transforming of second messengers. Methods：We chose adult female S-D rat AP culture 48h and added SP for different incubating time. After incubating Fura-2/AM was loaded as calcium fluorescent probe. The changes of Ca^2＋ in AP cells were observed by cation meaturement system. Results：The value of Ca^2＋ increased in a time-dependent manner with 100nmol/L SP from 10min to 30min while decreased at 40min and 50min. The value of Ca^2＋ was 211±16nmol/L,369±51nmol/L,358± 24nmol/L, 239±36nmol/L respectively from 20min to 50min. A significant difference was showed comparing to 10min group（117±17nmol/L）（P〈0. 05）. Conclusions：The system can accurately reflect the concentration of free Ca^2＋ in AP cells. Biological responses following activation of SPR are mediated by the second messenger Ca^2＋. The theory of SP controlling the reproducing axis is confirmed in the way of information transforming.