鸡血清极低密度脂蛋白对鸡颗粒细胞孕酮合成的影响

用鸡的颗粒细胞(granulosa cell,G·C·)进行极低密度脂蛋白(VLDL)对于孕酮合成的研究。按序列超速离心法分离鸡血清脂蛋白。用放射免疫法测定孕酮的量。实验分组:G·C·加绵羊促黄体生成素(OLH),其浓度范围为1—50 ng/ml,此为OLH组;G·C·加VLDL(最终浓度400μg/ml)再加OLH,此为VLDL组;G·C未加VLDL和OLH则为对照组。实验结果:(1).OLH组能促进G·C·孕酮的合成而且孕酮的生成量随着OLH量的增加而增加。(2)VLDL组孕酮生成量较OLH组显著增高,两者之间有显著性差异(P<0.001)。(3)用3次实验结果合并计算VLDL组和OLH组的平均数相当于对照组平均数的百分数,发现VLDL组明显高于OLH组。实验结果说明VLDL是携带胆固醇的脂蛋白,从而在OLH作用下,使颗粒细胞合成孕酮的量增多。     

化学发光法测定正常人皮肤纤维母细胞高密度脂蛋白(HDL)受体功能

A cbemiluminescence assay of human skin fibroblasts HDL receptor function was established.HDL3 was labelled with 6(N-(4-amino butyl)~N-ethyl)-amino-2,3-dihydro phthalazine l,4-dione(ABEI) by the carbodiimide(EDC).The labelled compound (HDL3-ABEI) was water soluble and non-toxic.It was stored at -30*********C before use.HDL3 was isolated by sequential ultracentrifugation.The HDL3 obtained was Apo-E-free.After incubation of HDL3 in waterbath at 52**********C for 3 hours, the HDL3 was labelled with ABEI to a ratio of 0.89 m mol ABEI/mmol HDL3.The HDL3-ABEI showed positive immunore action with antibodies of Apo A-********I and Apo A-*******II, indic-eting that the labelled HDL3 still retained its immunological characteristics.The conditions for determination of HDL receptor function of human skin fibroblasts were studied.The optimum quantity of HDL3-ABEI for combination with fibroblasts HDL receptors at 4癈 was 20********ug.The optimum time for combination of HDL3-ABEI with fibroblasts HDL receptors at 4*********C was 1 hour.The amount of HDL required for blocking HDL receptors was 400********ug.The optimum cell number for combination of HDL3-ABEI with HDL receptors was 1**********X105 cells/ml.In the mean time, the decrease in luminescence intensity with the increase in number of cells was observed.Again a high protein concentration would cause inhibition to the assay system.