益气温经方对CKIP-1介导的破骨细胞凋亡的影响

Effect of Yiqiwenjing Prescription on Osteoclasts Apoptosis Mediated by CKIP-1

目的:观察益气温经方对酪蛋白激酶2相互作用蛋白1(Casein Kinase 2Interaction Protein 1,CKIP-1)介导的破骨细胞凋亡程度的影响。方法:从SD乳鼠中分离破骨细胞体外培养,建立CKIP-1过表达和低表达载体,病毒包装构建稳定表达的细胞系。分为以下5组:破骨细胞对照组(A组),破骨细胞+空载病毒组(B组),破骨细胞+CKIP-1过表达病毒组(C组),破骨细胞+CKIP-1过表达病毒+益气温经方处理组(D组),破骨细胞+CKIP-1低表达病毒组(E组)。病毒感染96h后,Annexin VFITC/PI检测细胞凋亡。结果:在破骨细胞中,与A组相比,过表达CKIP-1病毒组凋亡细胞比例显著降低,差异有统计学意义(P=0.017),而益气温经方处理后,凋亡细胞比例有升高的趋势,与A组相比差异无统计学意义(P=0.059),与C组相比差异有统计学意义(P=0.000 6);与A组相比,低表达CKIP-1组凋亡细胞比例显著升高,差异有统计学意义(P<0.000 1)。结论:CKIP-1具有抑制破骨细胞凋亡的功能,益气温经方可能通过逆转这种趋势,抑制骨吸收,从而起到治疗骨质疏松的作用。

Objective：To observe the effect of Yiqiwenjing prescription on osteoclasts apoptosis mediated by casein kinase 2 interaction protein 1（CKIP-1）.Methods：Osteoclasts were isolated from SD neonatal rats in vitro,we established CKIP-1 overexpression and low expression vector.With the method of virus packaging,stable expressed cell lines were established.Then these cell lines were divided into the following 5 groups：group A,osteoclasts control group;group B,osteoclasts ＆vector group;group C,osteoclasts ＆ CKIP-1 overexpression virus group;group D,osteoclasts ＆ CKIP-1 overexpression virus ＆ Yiqiwenjing prescription intervened group;group E,osteoclasts ＆ CKIP-1 low expression virus group.96 hafter virus infection,Annexin V-FITC/PI was used to detect apoptosis.Results：Among osteoclasts,compared with the group A,the proportion of apoptotic cells in CKIP-1 overexpression virus group was significantly reduced（P=0.017）.After intervened by Yiqiwenjing prescription,the proportion of apoptotic cells increased,and there were no significant difference compared with group A（P =0.059）,while significant difference compared with group C（P =0.000 6）.The proportion of apoptotic cells showed significantly increased compared with group A（P〈0.000 1）.Conclusion：CKIP-1 has the function of inhibiting the apoptosis of osteoclasts,and Yiqiwenjing prescription’s effect may lie in reversing this trend,inhibiting bone absorption,and treating osteoporosis.