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中草药提取物对小鼠乳腺上皮细胞增殖的影响
王希春1, 周 凡1, 冯士彬1
安徽农业大学 动物科技学院
摘要:
【目的】研究中草药单味及复方提取物对原代培养的小鼠乳腺上皮细胞增殖的影响,旨在为探明中草药治疗奶牛乳腺疾病机制提供试验依据。【方法】采用组织块种植法培养3月龄、分娩后3~5 d的昆明种远交系小白鼠的乳腺组织,采用胰蛋白酶消化法获得相对纯化的小鼠乳腺上皮细胞,并运用SDS PAGE法进行鉴定。制备5味中草药(重楼、紫花地丁、木芙蓉、半枝莲、泽兰叶)及治疗奶牛乳房炎用中草药复方提取物,分别作用于对数生长期的乳腺上皮细胞5 d,用四甲基偶氮唑盐法(MTT法)分析细胞活力。【结果】将通过组织块种植法与胰蛋白酶消化法相结合,可得到纯化的有酪蛋白分泌功能的乳腺上皮细胞;中草药重楼、紫花地丁、木芙蓉、半枝莲、泽兰叶提取物在其质量浓度分别为8,40,40,1.6,8 mg/mL时,对小鼠乳腺上皮细胞增殖作用最强;除了紫花地丁,其他4种中草药提取物在其质量浓度为200 mg/mL时,对小鼠乳腺上皮细胞增殖的抑制作用最强。中草药复方提取物质量浓度为8~200 mg/mL时,对小鼠乳腺细胞有显著促进作用,随着质量浓度的增加,促增殖作用减弱。【结论】中草药单味及复方提取物在一定质量浓度下对乳腺上皮细胞增殖产生了促进或抑制作用,且中草药复方提取物的适用质量浓度范围较广。
关键词:  中草药  小鼠乳腺上皮细胞  细胞培养  增殖
DOI:
分类号:
基金项目:安徽省科技攻关项目(08010302157)
Effects of Chinese medicinal herb extracts on proliferation of mice mammary gland epithelial cells
Abstract:
【Objective】In order to explore the mechanism of treatment dairy cattle breast diseases by using Chinese medicinal herbs,the experiment was carried to study the effects of Chinese medicinal herb extracts on proliferation of normal mice mammary gland epithelial cells in primary culture.【Method】The mammary tissue clumps from KM mice in lactation period were obtained and cultured to harvest epithelial cells by both tissue transfer and trypsin digestion. Then the proliferation effects of the cells were detected by SDS PAGE.Five kinds of Chinese medicinal herbs (Rhizome paris,Herba violae,Folium hibisci mutabilis,Herba scutellaria barbata,Herba lycop)and the complex were extracted to add to cell cultural medium for five days.The vigor of mice mammary gland epithelial cells was determined by MTT.【Result】The result indicated that the normal mice mammary gland epithelial cells which had function of casein excretion could be purified by both tissue transfer and trypsin digestion;the mass concentration of Rhizome paris,Herba violae,Folium hibisci mutabilis,Herba scutellaria barbata,Herba lycop was 8,40,40,1.6,8 mg/mL,and Chinese medicinal herb extracts significantly promoted the proliferation of mice mammary gland epithelial cells;Except for Herba violae,the other four Chinese medicinal herbs extract significantly restrained proliferation of mice mammary gland epithelial cells.The mass concentration of Chinese medicinal herb complex extract was 8-200 mg/mL,the mice mammary gland epithelial cells can be enhanced significantly,and with the growth of mass concentration,the proliferation effect was decreased.【Conclusion】The experiment revealed that certain concentration of Chinese medicinal herbs and the complex exacts could promote or restrain mammary gland epithelial cells,and the complex had a wider range of mass concentration.
Key words:  Chinese medicinal herbs  mice mammary gland epithelial  cell culture  proliferation