巴戟天含药血清对成骨-破骨细胞共育体系 OPGmRNA、RANKLmRNA 表达的影响
Effect of morinda officinalis- containing serum on the mRNA expressions of OPG and RANKL in the osteoblast and osteoclast co-culture system
  
DOI:
中文关键词:  中医中药;巴戟天;成骨-破骨细胞共育体系;细胞核因子kB受体活化因子配体(RANKL)  破骨细胞抑制因子 (OPG,osteoprotegerin)  细胞核因子 kB 受体活化因子(RANK,receptor activator of NF-kB)
英文关键词:Chinese medcine and Chinese herb  Morinda officinalis  Osteoblast and osteoclast co-cultured system  RANKL  OPG
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黄慧' 2陈健 何剑全 郑素玉 张永晟 李艺敏1 1.福建中医药大学,福州350108 2. 厦门大学附属中山医院康复科,厦门361004 
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中文摘要:
      目的探讨不同浓度巴戟天含药血清对体外成骨-破骨细胞共育体系OPG、RANKLmRNA表达的影响方法取健康5 周龄SD大鼠12只,体重165 -172 g。随机分4组,分别予生理盐水及低中髙三种浓度巴戟天溶液灌胃,分离血清。取健康5 周龄SD大鼠6只,体重165 - 172 g,取四肢长骨骨髓基质细胞,诱导培养破骨细胞;用24 h内新生SD乳鼠4只,体重5 ~6 g, 头盖骨分离体外培养成骨细胞。将成骨细胞加人含有培养5 d的破骨细胞中,共育2 d后分别改用不同浓度巴戟天含药血清 与不含药血清的培养基培养,设低、中、髙浓度组及对照组,培养3d后提取各组总RNA,应用RT-PCR检测各组OPGmRNA、 RANKLmRNA表达。结果RT-PCR结果示,中、髙浓度组RANKLmRNA表达量均低于对照组(P < 0. 05),而OPGmRNA表达 量显著髙于对照组(P <0. 001)。结论巴戟天含药血清可上调共育体系中OPGmRNA表达,并下调RANKLmRNA表达,从而 降低破骨细胞分化成熟及骨吸收活性。
英文摘要:
      Objective To investigate the effect of the morinda officinalis-containing serum on the mRNA expressions of the OPG and RANKL in osteoblast and osteoclast co-culture system. Methods Twelve 5-week-old healthy SD rats (165 - 172g) were randomly assigned to 4 groups. Rats in each group were given different concentrations of morinda officinalis-containing serum or normal saline. The blood of the animals was extracted. Bone marrow cells were harvested from the femurs and tibiae of 6 5-week- old SD rats (165 - 172g). The cells were induced to osteoclasts. Osteoblasts were separated from the cranium of 4 24h newborn SD rats (5 - 6g). The osteoblasts were added to 5-day osteoclast culture. After 2-day co-culture ’ the different concentrations of morinda officinalis-containing serum or normal saline-containing serum were added to the culture plates. After intervention 3 days, total RNA was extracted from cells in high, middle, and low concentrations of morinda officinalis groups and control groups. The OPG and RANKL mRNA expressions were detected using real-time PCR. Results The results of RT-PCR indicated that RANKL mRNA expression was lower in all concentrations of morinda officinalis-containing serum groups than in the control group (P < 0.05),but OPG expression was higher than in the control group (P <0. 001). Conclusion Morinda officinalis-containing serum up-regulates the expression of OPG mRNA ’ but down-regulates the expression of RANKL mRNA in the co-culture system ’ leading to the decrease of osteoclast differentiation and activity of bone resorption.
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