温肾中药对原代培养正常人乳腺上皮细胞增殖的影响

目的观察温肾中药对雌二醇和孕酮所致的原代培养正常人乳腺上皮细胞增殖的影响。方法采用消化法进行正常人乳腺上皮细胞分离及原代培养。在相差倒置显微镜和透射电镜下对细胞进行形态学鉴定。应用雌二醇和孕酮干预以促使正常人乳腺上皮细胞增殖,给予不同浓度温肾中药及他莫昔芬进行药物干预。采用MTT法检测不同浓度、不同药物干预下乳腺上皮细胞的增殖情况。结果原代培养正常人乳腺上皮细胞经形态学鉴定符合正常人乳腺上皮细胞的形态学特征。浓度均为1×10-5g/L的雌二醇和孕酮混合培养液可以明显促进正常人乳腺上皮细胞增殖。高浓度温肾中药能明显促进原代培养正常人乳腺上皮细胞增殖,而低浓度温肾中药则可抑制其增殖。他莫昔芬对原代培养正常人乳腺上皮细胞增殖无明显影响。结论雌二醇和孕酮可以促进正常人乳腺上皮细胞增殖。温肾中药调节正常人乳腺上皮细胞增殖的作用与药物浓度相关。

Effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture

OBJECTIVE: To evaluate the effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture. METHODS: The normal human mammary epithelial cells were dissociated by digestion with collagenase type I. The morphological identification of normal human mammary epithelial cells in primary culture was determined under inverted phase contrast microscope and transmission electron microscope. The estradiol and progesterone were added to culture medium to induce the proliferation of normal human mammary epithelial cells in primary culture, and the different doses of Chinese herbal medicine for warming kidney and tamoxifen were also added into the culture medium. The MTT assay was used to evaluate the proliferation of normal human mammary epithelial cells in primary culture under the different conditions of interventions. RESULTS: The normal human mammary epithelial cells in primary culture presented typically morphological features of normal human mammary epithelial cells in vitro. The proliferation of normal human mammary epithelial cells in primary culture was significantly improved by intervention of estradiol and progesterone both with concentration of 1x10(-5) g/L. The high-dose Chinese herbal medicine for warming kidney could obviously promote the proliferation of normal human mammary epithelial cells in primary culture, while the proliferation was obviously inhibited by low-dose Chinese herbal medicine for warming kidney. The tamoxifen exerted no effects on the proliferation of normal human mammary epithelial cells in primary culture. CONCLUSION: The estradiol and progesterone can enhance the ability of proliferation of normal human mammary epithelial cells in primary culture. The regulating effect of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture is dose-dependent.