CAJ | 학술논문

背景：骨髓间充质干细胞中可能存在芳香化酶和雌激素受体相互作用的雌激素信号途径，调控骨髓间充质干细胞生物活性。 　　目的：观察成人骨髓间充质干细胞成骨诱导分化过程中芳香化酶和雌激素相关受体的表达。 　　方法：将骨髓间充质干细胞分为单纯培养组和成骨诱导组，分别用低糖DMEM完全培养基和成骨诱导分化完全培养基培养，MTT检测细胞增殖能力，茜素红染色观察成骨矿化能力，qPCR和Western blot检测芳香化酶、雌激素受体α、雌激素受体β和雌激素受体相关受体αmRNA和蛋白表达，ELISA检测细胞上清液和裂解液中雌二醇水平。 　　结果与结论：培养72 h，成骨诱导组增殖能力最强；培养21 d，成骨诱导组茜素红染色显示大量钙沉积；qPCR和Western blot结果显示成骨诱导组能促进芳香化酶和雌激素受体αmRNA和蛋白表达，抑制雌激素受体相关受体αmRNA和蛋白表达，成骨诱导组雌激素受体βmRNA和蛋白表达与单纯培养组无差异；ELISA结果显示成骨诱导组上清液中雌二醇水平高于裂解液及单纯培养组上清液中雌二醇水平。结果表明成人骨髓间充质干细胞及成骨诱导分化后能表达芳香化酶、雌激素受体α、雌激素受体β和雌激素受体相关受体α，亦能自身合成和分泌雌激素，由此产生的雌激素可能是通过相关受体对骨髓间充质干细胞成骨分化发挥作用。
배경：골수간충질간세포중가능존재방향화매화자격소수체상호작용적자격소신호도경，조공골수간충질간세포생물활성。 　　목적：관찰성인골수간충질간세포성골유도분화과정중방향화매화자격소상관수체적표체。 　　방법：장골수간충질간세포분위단순배양조화성골유도조，분별용저당DMEM완전배양기화성골유도분화완전배양기배양，MTT검측세포증식능력，천소홍염색관찰성골광화능력，qPCR화Western blot검측방향화매、자격소수체α、자격소수체β화자격소수체상관수체αmRNA화단백표체，ELISA검측세포상청액화렬해액중자이순수평。 　　결과여결론：배양72 h，성골유도조증식능력최강；배양21 d，성골유도조천소홍염색현시대량개침적；qPCR화Western blot결과현시성골유도조능촉진방향화매화자격소수체αmRNA화단백표체，억제자격소수체상관수체αmRNA화단백표체，성골유도조자격소수체βmRNA화단백표체여단순배양조무차이；ELISA결과현시성골유도조상청액중자이순수평고우렬해액급단순배양조상청액중자이순수평。결과표명성인골수간충질간세포급성골유도분화후능표체방향화매、자격소수체α、자격소수체β화자격소수체상관수체α，역능자신합성화분비자격소，유차산생적자격소가능시통과상관수체대골수간충질간세포성골분화발휘작용。
BACKGROUND:Estrogen signaling pathway for interaction between aromatase and estrogen-related receptor may exist in bone marrow mesenchymal stem cel s, which is used for regulating biological activity of bone marrow mesenchymal stem cel s. OBJECTIVE:To observe the expression of aromatase and estrogen-related receptors in adult bone marrow mesenchymal stem cel s during osteogenic differentiation. METHODS:Bone marrow mesenchymal stem cel s were respectively cultured in low-glucose DMEM medium (control group) and osteogenic induction medium (induction group). Cel proliferation and calcium deposition were determined by MTT assay and alizarin red staining, respectively. The expression of aromatase, estrogen receptorα, estrogen receptorβ, and estrogen-related receptorαduring osteogenic differentiation were determined by real-time PCR and western blot analysis. Estradiol levels in supernatants and lysates were detected by ELISA method. RESULTS AND CONCLUSION:In the induction group, the proliferation ability of bone marrow mesenchymal stem cel s was the strongest at 72 hours of culture;while there were a great amount of calcium nodules formed at 21 days of culture. Results from PCR and western blot assay showed that the expression of aromatase and estrogen receptorαwas improved in the induction group, but the expression of estrogen-related receptorαwas inhibited. There was no difference in the expression of estrogen receptorβbetween the two groups. ELISA results indicated that the level of estradiol in the supernatant of induction group was the highest. These findings indicate that aromatase, estrogen receptorα, estrogen receptorβand estrogen-related receptorαare al involved in osteogenesis of bone marrow mesenchymal stem cel s. Moreover, estradiol can be synthesized and secreted in bone marrow mesenchymal stem cel s, and most likely, promote the osteogenic differentiation of bone marrow mesenchymal stem cel s by related receptor pathway.