原代培养大鼠前列腺细胞建立前列腺增生筛药模型  

Establishment of a benign prostatic hyperplasia ( BPH) drug screening model with primary cultured rat prostatic cells in vitro

作　　者：吴建辉[1] 李军[2] 苏欣[1] 闫晗[1] 桂博[1] 蒋秀蓉[1] 孙祖越[1] 

机构地区：[1]上海市计划生育科学研究所,中国生育调节药物毒理检测中心,上海200032 [2]浦东新区人民医院泌尿科,上海201200

出　　处：《中国实验动物学报》2013年第5期10-14,I0002-I0004,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学青年基金项目(21007041);十二五"重大新药创制"科技重大专项(2011ZX09301-005);上海市科委研发公共服务平台项目(13DZ2291300);上海市人才发展基金资助项目(201372)

摘　　要：目的建立原代培养大鼠前列腺上皮细胞体外筛药模型。方法无菌状态下取雄性SD大鼠腹侧叶前列腺,称量后,剪成1 mm3小块,经Ⅱ型胶原酶消化1 h后,过滤、离心获取前列腺细胞,接种于24孔板培养并对细胞进行形态学和免疫组织化学鉴定。获取的大鼠前列腺上皮细胞分别接种于96孔板和24孔板培养12 d后给予系列剂量(0.1～1000μmol/L)的他莫昔芬和阳性药物爱普列特处理72 h,利用CCK-8法检测前列腺上皮细胞存活率并计算药物IC50值,并进一步通过Giemsa染色后观察细胞生长及形态变化。结果细胞鉴定结果显示,获取的细胞具典型上皮细胞形态特征,细胞角蛋白和前列腺特异性抗原表达阳性,提示所获细胞为前列腺上皮细胞。爱普列特与前列腺上皮细胞孵育72 h后,CCK-8法检测结果显示能够明显抑制前列腺上皮细胞生长,其IC50值为42.7μM,进一步镜下观察结果显示,爱普列特未明显改变大鼠前列腺上皮细胞形态,但能导致存活细胞数减少。他莫昔芬则对大鼠前列腺上皮细胞生长无明显抑制作用,镜下观察前列腺上皮细胞数量和形态未见明显改变。结论利用原代培养SD大鼠前列腺上皮细胞可以成功建立前列腺增生体外筛选模型。Objective To establish a drug screening model with primary cultured rat prostatic epithelial cells in vitro. Methods The ventral prostate was taken from adult SD rats under sterile conditions, weighted and cut into 1mm^3 pieces, and digested with collagenase 2 for 1 h. All prostate cells were harvested through filtering and centrifugation. The cells were seeded into 24-well plate and cultured for morphological and immunohistochemical examination. The prostatic epithelial cells were seeded into 96-well plate and 24-well plate, respectively, treated with epristeride（0. 1 -1000μmol/L） and tamoxifen （0. 1 - 1000 μmol/L）for 72 h after being cultured for 12 d. The survival rate of prostatic epithelial cells was detected by CCK-8 assay. The IC50 value of epristeride and tamoxifen was calculated, and their effects on the cell growth and morphology were further observed. Results According to cell morphology and immunohistochemical results, all the rat prostate cells displayed typical epithelial cell shape, and their expressions of cytokeratin and prostate specific antigens were positive, suggesting that the harvested cells were prostatic epithelial cells. After the prostate epithelial cells being treated with epristeride for 72 h, their growth was significantly inhibited, and the IC50 value of epristeride was 42.7 μmol/L. The morphological observation further showed that epristeride did not significantly change the cell shape, but reduced the num- ber of survivaing ceils, while tamoxifen did not inhibit the cell growth, and had no obvious effect on cell number and cell shape. Conclusion A benign prostatic hyperplasia drug screening model can be successfully established with primary cultured rat prostatic epithelial cells.

关 键 词：前列腺增生 模型 上皮细胞 原代培养 药物筛选 大鼠 

分 类 号：Q95-33[生物学—动物学]