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目的观察钾通道在SD大鼠前列腺上皮细胞增殖和凋亡中的作用。方法采用胶原酶消化SD大鼠前列腺组织建立原代SD大鼠前列腺上皮细胞株,角化蛋白免疫组织化学染色鉴定培养细胞,将SD大鼠前列腺上皮细胞分为对照组、钾通道阻断剂(四乙胺,TEA)组、开放剂(尼可地尔,Nicorandil)组,TEA的浓度为1、5、15mmol/L,Nicorandil的浓度为10、25、100μmol/L,各作用于细胞24、48、72h绘制细胞生长曲线,运用MTT方法、Hoechst33258细胞核染色法和流式细胞仪(FCM)观察传代后细胞增殖和凋亡状况。结果培养的细胞具有典型的上皮细胞形态特征,Pan-Keratin染色阳性,与对照组相比,MTT法和Hoechst33258染色法均显示,15mmol/L的钾离子通道阻滞剂TEA显著增加前列腺上皮细胞数(P<0.01);25μmol/L～100μmol/L/Nicorandil显著减少前列腺上皮细胞数(P<0.01),且FCM显示前列腺上皮细胞有明显的凋亡峰。Hoechst33258染色法显示细胞核固缩、裂解,呈现明显的凋亡形态特征。结论钾离子通道对体外培养的SD大鼠前列腺上皮细胞增殖与凋亡状态起重要的调节作用。
Objective To observe the role of potassium channels in the proliferation and apoptosis of prostate epithelial cells in SD rats. Methods The prostatic epithelial cell line of primary SD rat was established by collagenase digestion of rat prostate tissue. The cultured cells were identified by keratinized immunohistochemical staining. The prostate epithelial cells of SD rats were divided into control group, potassium channel blocker TEA group and Nicorandil group. The concentration of TEA was 1,5 and 15mmol / L, the concentration of Nicorandil was 10, 25 and 100μmol / L, The cell growth curve was drawn at 48 and 72h. The cell proliferation and apoptosis were observed by MTT assay, Hoechst 33258 nuclear staining and flow cytometry (FCM). Results Compared with the control group, MTT assay and Hoechst33258 staining showed that the cultured cells had the typical morphology of epithelial cells and positive staining of Pan-Keratin. TEA of 15mmol / L potassium channel blocker significantly increased the number of prostate epithelial cells (P <0.01). The number of prostatic epithelial cells (P <0.01) was significantly decreased from 25μmol / L to 100μmol / L / Nicorandil, and FCM showed significant apoptosis peak in prostatic epithelial cells. Hoechst33258 staining showed nuclear pyknosis and lysis, showing obvious morphological features of apoptosis. Conclusion Potassium channel plays an important role in regulating the proliferation and apoptosis of prostatic epithelial cells cultured in vitro in SD rats.