钾离子通道阻滞剂对SD大鼠前列腺上皮细胞的增殖作用

Potassium Channel Blocker Regulates the Proliferation of Prostatic Epithelial Cells in SD Rats

目的:观察钾离子通道阻滞剂四乙胺(TEA)、四氨基吡啶(4-AP)、格列本脲(Glib)对体外培养SD大鼠前列腺上皮细胞增殖的调节作用。方法:胶原酶消化组织建立原代SD大鼠前列腺上皮细胞株,角化蛋白免疫组织化学染色(Pan-keratin)鉴定培养细胞,分别加入TEA(1、5、10mmol/L)、4-AP(1、5、10mmol/L)、Glib(10、50、100mol/L),培养24、48、72h后计数并绘制细胞生长柱形图,运用MTT方法和Hoechst33258细胞核染色法观察传代后细胞生长状况。对照组不加钾离子通道阻制剂。结果:培养细胞具有典型的上皮细胞形态特征,Pan-Keratin染色阳性,MTT法和Hoechst33258染色法均显示随着TEA、4-AP及Glib浓度的递增,作用于细胞24、48、72h后,与对照组相比,前列腺上皮细胞有不同程度的增殖(P<0.05或P<0.01)。结论:钾离子通道阻滞剂对体外培养的SD大鼠前列腺上皮细胞有增殖作用。

Objective： To investigate the regulatory effect of potassium channel blocker（ tetraethylammonium [ TEA ], aminopyridine [4-AP], glibenclamide[ Glib] ）on the proliferation of SD rat prostatic epithelial cells in vitro. Methods： The primary culture was prepared by collagenase dissociation of minced prostatic tissues. Cells were cultured in serum-free prostate epithelial cell growth media and identified by immunocytochemical studies. TEA and 4-AP at the concentration of 1,5 and 10 mmol/L and Glib at the concentration of 10, 50 and 100 mol/L were added, and after 24, 48 and 72 hours of culturing, a cell column diagram was drawn and the cell number counted. The post-passage cell growth was observed by MTT assay and Hoechst33258 nucleus staining. Results： The cultured cells showed the typical morphological features of epithelia, with positive stain. MTT assay and Hoechst33258 staining showed that TEA, 4-AP and Glib at the increasing concentration effected different degrees of proliferation of prostatic epithelial cells after 24, 48 and 72 h（ P 〈 0.01 ）. Conclusion ： The potassium channel blocker is a direct physiological regulator of the proliferation of SD rat prostatic epithelial cells. Natl J Androl,2007,13 （ 2 ） ：138-142