续断对成骨细胞增殖、分化、凋亡和细胞周期的影响

THE INFLUENCE OF RADIX DIPSACI ON THE PROLIFERATION, DIFFERENTIATION, WITHERING AND CELL CYCLE OF OSTEOBLASTS

在动物实验的基础上 ,探讨续断对成骨细胞增殖、分化、凋亡、细胞周期和 Ⅰ 型前胶原αmRNA表达影响。根据文献方法培养大鼠成骨细胞株UMR 10 6 /0 1,采用3 H Tdr法检测细胞增殖 ,PNPP法、RIA法、茜素红染色法以及定量RT PCR法分别检测碱性磷酸酶活性、骨钙素活性、矿化结节形成和Ⅰ 型胶原αmRNA的表达 ,从而了解续断对成骨细胞增殖、分化的影响 ;FCM法检测细胞周期。结果显示续断中、高剂量能显著促进成骨细胞的增殖、增加碱性磷酸酶的表达及矿化结节形成的数量 ,促进成骨细胞骨钙素和Ⅰ 型前胶原mRNA的表达 (P <0 .0 1,P <0 .0 5 ) ,和雌激素组比较差异无显著性。续断高剂量组细胞增殖率、S期细胞比率和细胞增殖指数等均显著高于空白对照组 (P <0 .0 1,P <0 .0 5 ) ,和雌激素组比较差异也无显著性。细胞凋亡率和G0 G1期细胞比率显著低于空白对照组 (P <0 .0 1,P <0 .0 5 )。表明续断能有效促进成骨细胞的分化、增殖 ,防止成骨细胞凋亡 ,可能是该药促进骨折愈合、防治骨质疏松的机制之一。

The aim of the study is at studying the influence of Radix Dipsaci on the proliferation, differentiation, withering and cell cycle of osteoblasts and the type-I procollagen αmRNA expression based on the animal experiments. Rat osteoblast strains UMR-106/01 were cultivated by the method reported in the references; the cell proliferation was determined by 3H-Tdr; the ALP- and BGP- activity, mineralized node formation and type-I collagen mRNA expression were detected by PNPP, RIA, alizarin-red staining method and quantitative RT-PCR as to find out the influence of Radix Dipsaci on the proliferation and differentiation of osteoblasts; and the cell cycles were detected by FCM method. The results showed that the moderate and high doses of Radix Dipsaci function to promote the proliferation and BGP and type-I procollagen mRNA expression of osteoblasts and increase the ALP expression and the mineralized node formation number (P < 0.01, P < 0.05); the cell proliferation rate, the S-staged cell ratio and the cell proliferation increase index of the high dose group were much higher than those of the blank control group (P < 0.01, P < 0.05) with no significant difference from the estrogen group. The cell withering rate and the C0-C1-staged cell proportion were much lower than those of the blank control group (P < 0.01, P < 0.05), suggesting that Radix Dipsaci can effectively promote the differentiation and proliferation of osteoblasts and prevent the withering of osteoblasts, which may be one of the mechanisms for stimulating fracture healing, preventing and treating osteoporosis.