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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 effect of black bone vine on osteoclast differentiation and determine the active site. Methods: Osteoclasts were obtained from rabbit bone marrow cells induced by vitamin D3 (VD3). The osteoclasts were identified by morphological observation, hematoxylin and eosin (HE) staining and TRAP staining. The effect of low, medium and high doses (1, 10, 100 mg · L-1) on the activity of TRAP, the number of TRAP + Results: Both high-dose and high-dose ethyl acetate layers of nigrum nigrum saturated with n-butanol had a significant inhibitory effect on the differentiation of bone marrow cells into osteoclasts (P <0.01) and in a dose-dependent manner Sex. In the experiment of inhibiting the bone resorption of osteoclasts, all the dose groups of Eupatorium villosum all inhibited the bone resorption lacuna area (P <0.01), while the n-butyl alcohol group had significant Inhibition (P <0.01), all in a dose-dependent manner. CONCLUSION: The main active site that inhibits the differentiation of bone marrow cells into osteoclasts and the capacity of bone resorption is the ethyl acetate fraction.