黑骨藤抑制破骨细胞分化及骨吸收能力的活性部位研究

Research on Active Fraction of Periploca forrestii Inhibiting Osteoclast Differentiation and Bone Resorption Ability

目的:探究黑骨藤对破骨细胞分化的影响并确定其活性部位。方法:采用维生素D3(VD3)诱导兔骨髓细胞法获得破骨细胞,以形态学观察、苏木素和伊红(HE)染色、抗酒石酸酸性磷酸酶(TRAP)染色鉴定破骨细胞,然后通过测定黑骨藤各极性部位低、中、高剂量组(1,10,100 mg·L-1)对破骨细胞TRAP酶活力,TRAP+细胞计数及骨吸收陷窝面积的影响来确定活性部位。结果:黑骨藤水饱和正丁醇层的高剂量组及乙酸乙酯层的中、高剂量组均对骨髓细胞向破骨细胞分化具有显著的抑制作用(P<0.01),并具有剂量依赖性。在抑制破骨细胞骨吸收能力实验中,黑骨藤乙酸乙酯层各剂量组均有显著抑制骨吸收陷窝面积的作用(P<0.01),而正丁醇层中、高剂量组具有显著抑制作用(P<0.01),均呈剂量依赖性。结论:黑骨藤抑制骨髓细胞向破骨细胞分化及骨吸收能力的主要活性部位是乙酸乙酯部位。

Objective： To investigate the effects of Periploca forrestii against osteoclast differentiation and determine its active fraction. Method： The three indicators-morphological observation, hematoxylin and eosin （HE） staining, tartrate-resisstant acid phosphatase （TRAP） staining were used to identify osteoclasts which was obtained by vitamin D3 （VD3 ） inducing rabbit bone marrow ceils. The active fraction was determined, the inhibitory effects of each polarity of P. forrestii with different doses-low, medium and high dose groups （ 1, 10, 100 mg·L-1） on the TRAP enzyme activity of osteoclasts, TRAP＋ cells and bone resorption whirlpool area were measured. Result： In addition to the high-dose of water-saturated n-butanol layer, high-dose group as well as middle-dose of ethyl acetate layer also had significant inhibition on marrow cells differentiating into osteoclasts with dose-dependent manner （P 〈 0.01 ）. The measuring result of the bone resorption ability of osteoclast, showed that ethyl acetate layer inhibited bone resorption whirlpool area dose-dependently （P 〈 0. 01）, while the high-dose group of n-butanol layer also had apparent effects （P 〈 0. 01 ）. Conclusion ： Ethyl acetate fraction is the most active part of P. forrestii to suppress differentiation of bone marrow ceils into osteoclast and bone resorption capacity.