Objective: To study the effects of muskolibanum combination on the proliferation and differentiation of prostate stem cells.
Methods: We cultured prostate epithelial cells and urogenital sinus mesenchymal (UGSM) cells from 7－10 d old C57BL/6 mice and 16－18 d old pregnant C57BL/6 mice, transplanted the mixed suspension of the two types of cells under the kidney envelope of SCIDCB.17 male mice, and harvested the transplants 30 days later. We randomly divided the SCIDCB.17 mice into four groups to be treated intragastrically with musk (n = 8), olibanum (n = 8), musk+olibanum (n = 7), and normal saline (blank control, n = 8)) respectively, all for 14 days. Then we collected the kidney tissue for observation of the morphology of the glandular tubes and differentiation of different subsets of stem cells by HE staining and determination of the expressions and distribution of P63, CD133, CD117 and Sca1 by immunohistochemistry and Western blot.
Results: A system was successfully established for the isolation and mixed culture of Sca1 Lin+ CD49f+ (LSC) cells of prostate stem cells and UGSM cells of the mouse embryonic prostate. Immunohistochemistry showed positive expressions of P63, CD133, Sca1, and CD117 in the prostatic acinar epithelia and proved the presence of prostatic acinar epithelial structure in the transplants. Compared with the blank control group, the expressions of CD133, Sca1 and CD117 were significantly increased in the musk, olibanum, and musk+olibanum groups (P< 0.05), higher in the musk+olibanum than in the musk or olibanum group (P< 0.05), and their protein expressions were even more elevated in the musk+olibanum group (P< 0.01), with statistically significant difference from the olibanum group (P< 0.05).
Conclusions: The combination of musk and olibanum can improve the proliferation and differentiation of prostate stem cells.
目的: 研究麝香、乳香配伍处理对前列腺干细胞增殖分化的影响。方法: 选用7~10天龄的C57BL/6雄性幼鼠和16~18天龄的孕鼠，分别分离培养出前列腺上皮细胞和泌尿生殖窦间质(UGSM)细胞，并将二者混悬液移植到SCIDCB.17雄性小鼠的肾包膜下，30 d后收获移植物。将SCIDCB.17小鼠随机分为4组：麝香组、乳香组、麝香+乳香组、空白对照组，每组8只，按组别分别予麝香、乳香、麝香+乳香、生理盐水连续灌胃14 d后，取肾脏组织。通过HE染色形态学观察麝香配伍乳香对前列腺干细胞移植后腺管分化形成、各亚群干细胞分化状态影响，免疫组化和Western印迹检测前列腺干细胞抗原标志物P63、CD133、CD117、sca1的表达分布情况。结果: 建立了前列腺干细胞Lin Sca1+CD49f+（LSC）细胞及小鼠胚胎UGSM细胞的分离、混合培养体系；免疫组化结果示类前列腺腺泡上皮样结构P63、CD133、Sca1、CD117染色呈阳性，证实移植物中可见前列腺腺泡上皮结构，且乳香组、麝香组的表达较空白对照组均有不同程度的增高,其中麝香+乳香组CD133、Sca1、CD117免疫组化的积分光密度（IOD）值均显著高于单用乳香组和空白对照组（均< 0.05），同时Sca1的表达水平也显著高于单用麝香组（P< 0.05），而P63的表达水平4组间无显著差异。Western印迹结果也显示，麝香+乳香组CD133、Sca1、CD117的相对表达量均显著高于单用乳香组和空白对照组（P< 0.05或< 0.01），Sca1的表达水平也显著高于单用麝香组（P< 0.01）。 结论: 麝香、乳香配伍使用能促进前列腺干细胞的增殖再生。.
Keywords: musk; mice; olibanum; prostate stem cell; urogenital sinus mesenchymal cell.