异烟肼、利福平联合应用对体外培养成骨细胞生物活性的影响  

Effects of different concentrations of Isoniazide and Rifampin on the bioactivity of cultured osteoblasts in vitro

作　　者：马永海 巩栋 王伟[1] 邵陇龙 甄平[3] MA Yonghai;GONG Dong;WANG Wei;SHAO Longlong;ZHEN Ping(Ningxia Medical University, Yinchuan 750000;Gansu University of Chinese Medicine, Lanzhou 730000;Department of Orthopedics, Lanzhou General Hospital of Lanzhou Military Region of PLA, Lanzhou 730050, China)

机构地区：[1]宁夏医科大学,宁夏银川750000 [2]甘肃中医药大学,甘肃兰州730000 [3]解放军兰州军区兰州总医院骨科,甘肃兰州730050

出　　处：《中国骨质疏松杂志》2019年第2期141-145,151,共6页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金(81371983)

摘　　要：目的研究不同浓度的异烟肼(isoniazid,INH)、利福平(rifampin,RFP)联合应用对大鼠成骨细胞(osteoblast,OB)增殖、碱性磷酸酶(alkaline phosphatase,ALP)活性、I型胶原蛋白表达和成骨细胞成熟与矿化能力的影响。方法分离、培养新生SD大鼠乳鼠颅骨OB,分别采用CCK-8法、碱性磷酸酶试剂盒检测ALP活性、免疫荧光染色法和茜素红染色法检测,观察不同浓度INH+RFP [(10+3)μg/mL、(20+6)μg/mL、(30+12)μg/mL、(40+24)μg/mL、(50+48)μg/mL]对成骨细胞增殖、ALP活性、Ⅰ型胶原蛋白表达以及矿化能力的影响。结果与对照组相比,当INH+RFP的浓度在(20+6)μg/mL及以上时,抑制大鼠OB增殖、I型胶原合成、使ALP活性及矿化能力显著下降(P<0.05),且均随着浓度增加而加强抑制作用。结论 INH、RFP联合用药较低浓度对成骨细胞有抑制作用,因此,骨关节结核病灶清除术后,局部用药应该严格控制药物浓度。Objective To investigate the effect of different concentrations of isoniazid(INH) and rifampicin(RFP) on cell proliferation, alkaline phosphatase(ALP) activity, expression of typeⅠcollagen and mineralization of osteoblasts derived from newly born SD rat in vitro. Methods Osteoblasts derived from newly born(<24 h) SD rats’ calvarias were cultured with INH and rifampicin at different concentrations of 10+3 μg/mL, 20+6μg/mL, 30+12μg/mL, 40+24μg/mL, 50+48μg/mL for some days, then evaluated for cell proliferation, ALP activity, expression of typeⅠcollagen and mineralization using CCK-8, ALP activity assay, immunofluorescence staining and alizarin red staining assay. Results When the concentration of INH and RFP reached 20+6 μg/mL and above, CCK-8 values, ALP activity, type I collagen synthesis and mineralization of OB were significantly decreased compared with those of the control group(P<0.05). With the increase in the concentration of INH and RFP, the inhibition increased gradually. Conclusion Our study shows that INH and RFP at lower concentration could inhibit osteoblasts. After the removal of bone and joint tuberculosis, it is necessary to strictly control the concentration of INH and RFP.

关 键 词：异烟肼 利福平 成骨细胞 碱性磷酸酶 Ⅰ型胶原 

分 类 号：R965.2[医药卫生—药理学]