摘要
目的研究吗啡对体外培养人绒毛膜滋养层细胞雌激素受体(ER)和孕激素受体(PR)表达的影响,探讨吗啡对滋养层细胞的直接作用,为进一步寻求减少或阻断吗啡依赖对胎盘组织的负面影响提供一定的理论依据。方法用胰酶-胶原酶消化法从人早孕绒毛组织中分离培养滋养层细胞,用免疫组织化学方法鉴定滋养层细胞;制备不同浓度的吗啡及其拮抗剂纳络酮,作用于体外培养的早孕绒毛膜滋养层细胞上,通过反转录-聚合酶链反应(RT-PCR)法半定量分析不同浓度的吗啡及其拮抗剂纳络酮对滋养层细胞ER mRNA、PR mRNA表达的影响。结果用改良法分离的人滋养层细胞不同浓度吗啡组滋养层细胞上ER mRNA表达与对照组比较有显著性下降(P<0.05),且与吗啡浓度呈负相关(r=-0.996,P<0.01);不同浓度吗啡组滋养层细胞上PR mRNA表达与对照组比较均有显著性下性(P<0.05),且与吗啡浓度呈负相关(r=-0.983,P<0.01)。结论用改良法进行人滋养层细胞培养,简捷易行,可培养大量高纯度人滋养层细胞供研究使用;从mRNA水平证明了吗啡可以引起体外培养的人早孕绒毛膜滋养层细胞内ER、PR数量下调,且与吗啡浓度呈负相关,可剂量依赖下调人滋养层细胞ER mRNA、PR mRNA的表达。
Objective To culture and identify the functions of digestion-isolated human trophoblast cells during first-trimester pregnancy, and to evaluate the effect of morphine on expressions of estrogen receptor （ER） and progesterone receptor （PR） mRNAs of trophoblast cells, so as to explore the direct actions of morphine on trophoblast cells and provide a theoretical basis for minimizing or blocking the negative impacts on placenta from morphine dependence, Methods The trophoblast cells were isolated from human chorionic villi during the first-trimester pregnancy by trypsin-collagenase digestion and functionally identified by microscopy and immunocytochemical staining. Morphine and naloxone in various concentrations were prepared and used to act on cultured trophoblast cells. Expressions of ER and PR mRNAs of trophoblast cells were examined with RT-PCR. Results Expression of ER mRNA on morphinetreated human trophoblast ceils was significantly down-regulated as compared with controls （P〈0.05） and negatively correlated with the level of morphine （r=-0.996, P〈0.01）. Expression of PR mRNA on morphine-treated human tro- phoblast cells was significantly down-regulated as compared with controls （P〈0.05） and negatively correlated with the level of morphine （r=-0.983, P〈0.01）. Conclusion The modified approach for trophoblast cell cuhure appeared easy to use and effective for massive cultivation of high-purity cells as needed in studies. At the mRNA level, morphine was demonstrated to down-regulate, negatively correlated with its concentration, the expression of ER and PR mRNAs of cultured human trophoblast cells during first-trimester pregnancy, suggesting a dose-dependent manner.
出处
《中国药物与临床》
CAS
2010年第4期397-399,共3页
Chinese Remedies & Clinics
关键词
吗啡
滋养层
受体
雌激素
受体
孕激素
Morphine
Trophoblasts
Receptor, estrogen
Receptor, progesterone
