摘要
目的探讨不同浓度巴戟天含药血清对体外培养成骨-破骨细胞共育体系中碳酸酐酶Ⅱ(CAⅡ)、活化T细胞核因子(NFAT2)mRNA表达的影响。方法取24 h内新生SD乳鼠头盖骨分离培养成骨细胞,取5周龄SD大鼠四肢长骨骨髓基质细胞,加入集落细胞刺激因子(M-CSF)和细胞核因子κB受体活化因子配体(RANKL)诱导培养破骨细胞。采用ALP染色鉴定成骨细胞,TRAP染色、骨吸收陷窝甲苯胺蓝染色、电镜等扫描鉴定破骨细胞,体外建立成骨-破骨细胞共育体系,设置高、中、低3种浓度巴戟天含药血清组和对照组,干预3d后提取各组总RNA,应用Real Time PCR(RT-PCR)方法测定各组CAⅡ、NFAT2mRNA表达并进行统计学分析。结果不同浓度巴戟天含药大鼠血清对成骨-破骨细胞共育体系CAⅡ、NFAT2 mRNA的表达均有抑制作用,且其抑制作用表现出一定的浓度依赖性;各组间差异有统计学意义(P<0.05)。结论巴戟天含药血清可抑制成骨-破骨细胞共育体系CAⅡ、NFAT2mRNA表达,从而达到降低破骨细胞分化成熟及骨吸收活性。
Objective To investigate the effect of different concentrations of morinda officinalis （RMO）- containing serum on the expressions of the carbonic anhydrase II （CAII） and nuclear factor 2 of activated T ceils （NFAT2） mRNA in osteoblast-osteoclast co-culture system. Methods Osteoblasts were isolated from the cranium of 24-hour newborn SD rats and cultured. Bone marrow cells were isolated from the femurs and the tibiae of 5-week-old SD rats. Then bone marrow cells were induced to osteoclast by addition of M-CSF and RANKL. Osteoblast ceils were identified using alkaline phosphatase （ALP） staining. Osteoclast cells were identified using tartrate resistant acidphos phatase （TRAP） staining, bone resorption lacuna toluidine blue staining, and electronic speculum. Osteoblast-osteoclast co-culture system was established in vitro. Study groups including groups with high, middle, and low concentrations of RMO-containing serum, and control group were established. Total RNA of the cells in each group was extracted after 3-day the intervention. The expression of CAII and NFAT2 mRNA was tested using real-time polymerase chain reaction （RT-PCR）. Results Different concentrations of RMO-containing serum had inhibitory effect on the expression of CAII and NFAT2 mRNA. And the inhibitory effect was concentration-dependent to some extent. The difference among different groups was significant （P 〈 0. 05 ）. Conclusion The RMO- containing serum can inhibit the expression of CAII and NFAT2 mRNA in osteoblast-osteoclast co-culture system. And it can also reduce the differentiation of osteoclasts and the activity of bone resorption.
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2013年第2期120-124,134,共6页
Chinese Journal of Osteoporosis
基金
厦门市科技局资助项目(NO.3502z20104031)
关键词
巴戟天
成骨一破骨细胞共育体系
碳酸酐酶Ⅱ
活化T细胞核因子2
Morinda officinalis
Osteoblast and osteoclast co -culture system
Carbonic anhydrase I1
Nuclear factor 2 of activated T cells
