CAJ | 학술논문

组织构建%骨组织工程%双膦酸盐%破骨细胞%抗酒石酸酸性磷酸酶%组织蛋白酶K%骨吸收陷窝%免疫印迹%免疫荧光化学%骨质疏松%扫描电镜%牙本质磨片%国家自然科学基金組織構建%骨組織工程%雙膦痠鹽%破骨細胞%抗酒石痠痠性燐痠酶%組織蛋白酶K%骨吸收陷窩%免疫印跡%免疫熒光化學%骨質疏鬆%掃描電鏡%牙本質磨片%國傢自然科學基金
조직구건%골조직공정%쌍련산염%파골세포%항주석산산성린산매%조직단백매K%골흡수함와%면역인적%면역형광화학%골질소송%소묘전경%아본질마편%국가자연과학기금
tissue engineering%diphosphonates%osteoclasts%osteoporosis

背景：有研究表明双膦酸盐可抑制破骨细胞的骨吸收功能，但对其骨吸收功能关键细胞因子组织蛋白酶K是否产生作用，至今少有报道。 　　目的：观察双膦酸盐对破骨细胞分化中组织蛋白酶K及骨吸收功能影响。 　　方法：用小鼠单核巨噬细胞RAW264.7诱导培养破骨细胞。实验分2组：对照组加入质量浓度100μg/L核因子κB受体活化因子配体进行诱导至收获细胞，双膦酸盐组在对照组的基础上加入10-7 mol/L阿仑膦酸盐处理至收获细胞。培养第7天检测各组破骨细胞生成和骨吸收功能，培养72 h免疫荧光检测两组组织蛋白酶K表达差异，Western blot检测组织蛋白酶K蛋白表达情况。 　　结果与结论：两组均有抗酒石酸酸性磷酸酶阳性多核破骨细胞生成，并在牙本质磨片上形成吸收陷窝；但对照组抗酒石酸酸性磷酸酶阳性多核细胞数目、吸收陷窝数目及陷窝面积均大于双膦酸盐组(P<0.01)。免疫荧光检测组织蛋白酶K表达对照组强于双膦酸盐组(P<0.01)；Western blot检测组织蛋白酶K表达双膦酸盐组低于对照组(P<0.01)。结果证实，双膦酸盐通过抑制组织蛋白酶K因子的表达，阻碍破骨细胞的骨吸收功能。
배경：유연구표명쌍련산염가억제파골세포적골흡수공능，단대기골흡수공능관건세포인자조직단백매K시부산생작용，지금소유보도。 　　목적：관찰쌍련산염대파골세포분화중조직단백매K급골흡수공능영향。 　　방법：용소서단핵거서세포RAW264.7유도배양파골세포。실험분2조：대조조가입질량농도100μg/L핵인자κB수체활화인자배체진행유도지수획세포，쌍련산염조재대조조적기출상가입10-7 mol/L아륜련산염처리지수획세포。배양제7천검측각조파골세포생성화골흡수공능，배양72 h면역형광검측량조조직단백매K표체차이，Western blot검측조직단백매K단백표체정황。 　　결과여결론：량조균유항주석산산성린산매양성다핵파골세포생성，병재아본질마편상형성흡수함와；단대조조항주석산산성린산매양성다핵세포수목、흡수함와수목급함와면적균대우쌍련산염조(P<0.01)。면역형광검측조직단백매K표체대조조강우쌍련산염조(P<0.01)；Western blot검측조직단백매K표체쌍련산염조저우대조조(P<0.01)。결과증실，쌍련산염통과억제조직단백매K인자적표체，조애파골세포적골흡수공능。
BACKGROUND:Studies have shown that bisphosphonates inhibit osteoclast resorption, but whether cathepsin K, a key cytokine of bone resorption, plays an effect has rarely been reported. OBJECTIVE:To study the effect of bisphosphonate on capthesin K and bone resorption function during osteoclast differentiation. METHODS:Osteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groups:control group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture and gene expression of capthesin K was detected by immunofluorescence method at 72 hours of culture. Western blot assay was used to detect capthesin K protein expression at 72 hours of culture. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group (P<0.01). Immunofluorescence showed expression of capthesin K was higher in the control group than the alendronate group (P<0.01);furthermore, the protein expression of capthesin K was also lower in the alendronate group than the control group (P<0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting gene expression of capthesin K.