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目的:确定清络通痹颗粒(QLT)含药血清的制备方法并观察其对体外培养破骨细胞的活性和骨吸收的影响。方法:选用高效液相色谱外标一点法测定,QLT含药血清中青藤碱的含量以确定含药血清的制备条件;乳鼠破骨细胞1×108/mL加入96孔板,将QLT 3.6,7.2,14.4 g.kg-1给药的大鼠含药血清1∶10稀释,分别加入各孔培养48 h,观察其对破骨细胞增殖、抗酒石酸酸性磷酸酶(TRAP)活性及破骨细胞形成的骨吸收陷窝的影响。结果:QLT 7.2,14.4 g.kg-1给药的含药血清可明显抑制破骨细胞的增殖能力和吸收陷窝面积(P<0.05,P<0.01),QLT 3.6,7.2,14.4 g.kg-1给药的含药血清可明显抑制破骨细胞的TRAP活性(P<0.05,P<0.01)。结论:QLT含药血清可抑制破骨细胞增殖、TRAP活性和骨吸收能力,对破骨细胞引起的骨质破坏产生抑制作用。
Objective: To determine the preparation method of Qingluotongbi granule (QLT) containing serum and observe its effect on osteoclast activity and bone resorption in vitro. Methods: The content of sinomenine in QLT-containing serum was determined by one-point HPLC method to determine the preparation conditions of serum containing serum. The osteoclast of 1 × 108 / mL was added into 96-well plate, and QLT 3.6 , 7.2, 14.4 g.kg-1 rat drug-containing serum 1:10 dilution, were added to each well for 48 h culture, observe its osteoclast proliferation, tartrate-resistant acid phosphatase (TRAP) activity and osteoclasts The cells that form the bone absorb the effects of the lacuna. Results: The serum containing QLT 7.2 and 14.4 g.kg-1 could significantly inhibit the proliferation and resorption lacunae of osteoclasts (P <0.05, P <0.01), QLT 3.6, 7.2 and 14.4 g.kg -1 could significantly inhibit the TRAP activity of osteoclasts (P <0.05, P <0.01). CONCLUSION: QLT-containing serum can inhibit osteoclast proliferation, TRAP activity and bone resorption ability, and inhibit osteoclast-induced bone destruction.