右归饮对假体周围骨溶解大鼠模型体外培养破骨细胞分化和相关基因表达的影响  

Effects of Yougui Drink on the Phantom around the Osteolysis Big Mouse Model in vitro Raise Osteoclast Differentiation and the Related Gene Expression Influence

作　　者：蔡运火[1] 鲍航行[1] 王金法[1] 单乐天[1] 杜文喜[2] 赵鹏[1] 方剑利[1] 肖鲁伟[1] 童培建[2] 

机构地区：[1]浙江中医药大学,杭州310053 [2]浙江中医药大学附属第一医院

出　　处：《浙江中医药大学学报》2012年第1期55-60,共6页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金(30873276)~~

摘　　要：[目的]为进一步观察右归饮对假体周围骨溶解模型大鼠体外诱导培育破骨细胞的分化以及相关基因表达的影响。[方法]制备假体周围骨溶解大鼠模型,六周后(造模)处死,无菌条件下用造模完成的大鼠的四肢骨髓腔中分离破骨细胞,接种于象牙薄片底物上,然后随机分为4组:高、中、低浓度右归饮血清模型组(Y1、Y2、Y3)和生理盐水对照血清模型组(Y4),分别添加高、中、低浓度右归饮含药血清及生理盐水对照血清;没有造模的大鼠的破骨细胞培养设为空白对照组(Y5),于第7d终止培养,通过抗酒石酸酸性磷酸酶(TRAP)染色对TRAP阳性多核破骨细胞计数;采用吖啶橙荧光染色法计数破骨细胞的凋亡率;用RT-PCR测定ATP6i、IL-1、IL-6基因相对表达量,分析破骨细胞骨吸收活性。[结果]高、中、低浓度右归饮血清模型组(Y1、Y2、Y3)与生理盐水血清模型组(Y4)相比,TRAP染色阳性的破骨细胞数量显著减少,破骨细胞的凋亡率明显增加,ATP6i、IL-1、IL-6基因表达明显减少。生理盐水血清模型组(Y4)与生理盐水血清空白组(Y5)相比,TRAP染色阳性的破骨细胞数量显著增加,破骨细胞的凋亡率无显著性差异(P>0.05)。[结论]一定浓度的右归饮含药血清对体外培养的颗粒病模型大鼠破骨细胞的增殖和分化具有明显的抑制作用,能显著抑制破骨细胞相关基因的表达。[Objective] To further observe Youguiyin to the bone around the prosthesis dissolution big mouse model in vitro induction cultivation osteoclast＇s differentiation as well as the correlation gene expression influence.[Method] Prepare the bone around the prosthesis dissolution big mouse model,six weeks later（makes mold） to execute,under the aseptic condition,make in the big mouse＇s four limbs medullary cavity of bones which the mold complete to separate the osteoclast,vaccinate on the ivory thin slice substrate,then divide into 4 groups stochastically： High,middle,the low concentration Youguiyin the blood serum model group（Y1,Y2,Y3） and the physiological saline comparison blood serum model group（Y4）,add separ-ately high,middle and low concentration Youguiyin including the medicine blood serum and the physiological saline comparison blood serum;Not make mold＇s big mouse＇s osteoclast raise to suppose for the blank control group（Y5）,in the 7d termination raise,dye through anti-tartaric acid acid phosphatase（TRAP） to the TRAP masculine gender polynuclear osteoclast counting;Use acridine orange fluorescence dyeing counting osteoclast＇s weak perishing rate;Determine ATP6i,IL-1,the IL-6 gene relative expression quantity with RT-PCR,analyse osteoclast bone absorption activeness.[Result] High,middle,low concentration Youguiyin the blood serum model groups（Y1,Y2,Y3） to compare with the physiological saline blood serum model group（Y4）,TRAP dyeing masculine osteoclast quantity obviously reduced,osteoclast＇s weak perishing rate increases obviously,ATP6i,IL-1,IL-6 gene expression obvious reduction.The physiological saline blood serum model group（Y4） compared with the physiological saline blood serum blank group（Y5）,TRAP dyeing masculine osteoclast quantity obviously increases,osteoclast＇s weak perishing rate non-significance difference.[Conclusion] Certain density Youguiyin has the obvious inhibitory action including the medicine blood serum to vitro raise pellet sickness model

关 键 词：右归饮 破骨细胞 颗粒病模型 细胞培养 

分 类 号：R331[医药卫生—人体生理学]