摘要
目的 观察薯蓣皂苷对破骨细胞(OC)分化及骨吸收功能的调控作用,并探讨可能的机制。方法 取对数期RAW 264.7细胞,分为对照组、低剂量组、中剂量组、高剂量组(加入薯蓣皂苷0,1,3,9μmol·mL^(-1))、高剂量+茴香霉素组(薯蓣皂苷9μmol·mL^(-1)、茴香霉素2 000 mg·L^(-1))。诱导分化第1,7天进行抗酒石酸酸性磷酸酶(TRAP)染色观察OC分化情况;以蛋白质印迹法检测OC特异性蛋白组织蛋白酶K(CTSK)蛋白表达;以牛骨片实验检测骨吸收功能;以蛋白质印迹法检测p38丝裂原活化蛋白激酶(p38 MAPK)、p-p38 MAPK、细胞外信号调节激酶(ERK1/2)、p-ERK1/2蛋白表达。结果 对照组、低剂量组、中剂量组、高剂量组、高剂量+茴香霉素组OC数目分别为(142.20±25.41),(115.40±22.63),(74.80±9.51),(23.80±4.16),(62.60±7.52)个;这5组的CTSK蛋白表达量分别为0.62±0.07,0.50±0.06,0.39±0.04,0.24±0.03,0.35±0.05;这5组的骨吸收陷窝面积分别为(2.41±0.56),(1.26±0.30),(0.85±0.12),(0.31±0.05),(0.64±0.09)mm;这5组的p-p38 MAPK/p38 MAPK分别为0.72±0.08,0.50±0.06,0.38±0.04,0.19±0.03,0.27±0.04,p-ERK1/2/ERK1/2分别为0.69±0.07,0.58±0.06,0.42±0.05,0.28±0.03,0.36±0.04。上述指标,低、中、高剂量组与对照组比较,中剂量组与低剂量组比较,高剂量组与中剂量组比较,高剂量+茴香霉素组与高剂量组比较,差异均有统计学意义(均P<0.05)。结论 薯蓣皂苷可抑制OC分化及骨吸收功能,且呈浓度依赖性,推测其作用机制与抑制p38MAPK信号通路有关。
Objective To observe the regulation effect of dioscin on osteoclast (OC) differentiation and bone resorption,and to explore the possible mechanism.Methods RAW 264.7 cells in log phase were divided into control group,low-dose group,medium-dose group,high-dose group (adding dioscin 0,1,3,9μmol·m L^(-1)),highdose+anisomycin group (dioscin 9μmol·m L^(-1),anisomycin 2 000mg·L^(-1)).OC differentiation on the 1st and 7th day of induction were observed by tartrate-resistant acid phosphatase (TRAP) staining.The expression of OC-specific protein cathepsin K (CTSK) protein were detected by Western blotting.The bone resorption function were detected by bovine bone slice experiment.The expression of p38 mitogenactivated protein kinase (p38 MAPK),p-p38 MAPK,extracellular signal-regulated kinase (ERK1/2),and p-ERK1/2 protein expression were detected by Western blotting.ResultsThe number of OC in control group,low-dose group,medium-dose group,high-dose group and high dose+anisomycin group were 142.20±25.41,115.40±22.63,74.80±9.51,23.80±4.16,62.60±7.52,the expression levels of CTSK protein in the 5 groups were 0.62±0.07,0.50±0.06,0.39±0.04,0.24±0.03,0.35±0.05,the bone resorption lacuna area in the 5 groups were (2.41±0.56),(1.26±0.30),(0.85±0.12),(0.31±0.05),(0.64±0.09) mm;,the p-p38 MAPK/p38 MAPK in the 5 groups were 0.72±0.08,0.50±0.06,0.38±0.04,0.19±0.03,0.27±0.04,p-ERK1/2/ERK1/2 were 0.69±0.07,0.58±0.06,0.42±0.05,0.28±0.03,0.36±0.04.For the above indicators,the differences between low-,medium-,highdose groups and control group,between medium-dose group and low-dose group,between high-dose group and medium-dose group,between high dose+anisomycin group and high-dose group were all statistically significant (all P<0.05).Conclusion Dioscin can inhibit OC differentiation and bone resorption in a concentration-dependent manner.It is speculated that its mechanism of action is related to the inhibition of p38MAPK signaling pathway.
作者
陈学武
王弘
CHEN Xue-wu;WANG Hong(Department of Spine Orthopedics,Wannan Medical College,Yijishan Hospital,Wuhu 241000,Anhui Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2022年第6期523-527,共5页
The Chinese Journal of Clinical Pharmacology
基金
安徽省自然科学基金资助项目(1308085MHl52)。
