[1]杨依然,刘钟,王均华,等.强骨饮对CKIP-1过表达成骨细胞的影响研究[J].中国中医骨伤科杂志,2018,26(10):1-5.
　YANG Yiran,LIU Zhong,WANG Junhua,et al.Effect of Qiangguyin on Rat Osteoblasts in CKIP-1 Overexpressing[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2018,26(10):1-5.
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强骨饮对CKIP-1过表达成骨细胞的影响研究()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第26卷
期数:
2018年10期
页码:
1-5
栏目:
实验研究
出版日期:
2018-10-10

文章信息/Info

Title:
Effect of Qiangguyin on Rat Osteoblasts in CKIP-1 Overexpressing
文章编号:
1005-0205(2018)10-0001-05
作者:
杨依然1; 刘钟1; 王均华1; 刘康2; 史晓林2△
1浙江中医药大学(杭州,310053) 2浙江中医药大学附属第二医院
Author(s):
YANG Yiran1; LIU Zhong1; WANG Junhua1; LIU Kang2; SHI Xiaolin2△
1Zhejiang University of Traditional Chinese Medicine,Hangzhou 310053,China; 2The Second Hospital Affiliated to Zhejiang University of Traditional Chinese Medicine,Hangzhou 310005,China.
关键词:
酪蛋白激酶2相互作用蛋白1;  成骨细胞;  过表达;  细胞增殖;  中药疗法
Keywords:
casein kinase 2 interaction protein 1;  osteoblast;  overexpression;  cell proliferation;  Chinese herbal medicine
分类号:
R-33
文献标志码:
A
摘要:
目的:观察强骨饮对酪蛋白激酶 2 相互作用蛋白1(CKIP-1)过表达慢病毒感染的体外小鼠成骨细胞的调控情况,研究强骨饮治疗骨质疏松的分子作用机制。方法:从2 d龄SD乳大鼠分离成骨细胞,构建CKIP-1过表达慢病毒,将成骨细胞分成4组:a组为空白成骨细胞对照组,b组为空载慢病毒感染的成骨细胞,c组为CKIP-1过表达慢病毒感染的成骨细胞,d组为CKIP-1过表达慢病毒感染成骨细胞+强骨饮处理。qRT-PCR检测成骨抑制基因CKIP-1和成骨相关基因ALP及RUNX2的表达,CKK-8检测细胞相对数目,茜素红染色鉴定成骨细胞,低倍光镜下观察钙化结节计数。结果:qRT-PCR检测:相比a组、b组,c组的CKIP-1有较高表达,差异有统计学意义(P<0.01),RUNX2和ALPL表达水平极低,差异有统计学意义(P<0.01); c组与d组比较,d组的CKIP-1表达水平显著降低,差异有统计学意义(P<0.01),RUNX2和ALPL表达显著升高,差异有统计学意义(P<0.01)。CKK-8检测:相比a组、b组,c组的细胞增殖水平显著降低,差异有统计学意义(P<0.01); 相比c组,d组强骨饮处理后,细胞增殖比例有升高的趋势,且差异有统计学意义(P<0.01)。茜素红染色:相比a组、b组,c组钙化结节明显减少; 相比c组,d组钙化结节显著增多。结论:CKIP-1过表达对成骨细胞的增殖、成骨分化及矿化有明显抑制作用,强骨饮能提高CKIP-1过表达病理状态下成骨细胞的活性和成骨分化及矿化能力。
Abstract:
Objective:To observe the effect of Qiangguyin on osteoblasts of mice infected by lentivirus overexpressing casein kinase 2 interaction protein 1(CKIP-1),and to study the molecular mechanism of Qiangguyin for treating osteoporosis.Methods:Osteoblasts were isolated from 2-day-old SD lactating rats to construct CKIP-1 over-expressed lentivirus.Osteoblasts were divided into 4 groups.a group,blank osteoblasts control group; b group,empty-loaded osteoblasts infected by lentivirus; c group,osteoblasts infected by lentivirus overexpression CKIP-1; d group,CKIP-1 overexpression osteoblasts infected by lentivirus combined with Qiangguyin treatment.The expression of CKIP-1,ALP and RUNX2 were detected by qRT-PCR,the relative number of osteoblasts was detected by CKK-8,the osteoblasts were identified by alizarin red staining,and the number of calcified nodules was observed under low power microscope.Results:The qRT-PCR assay showed:compared with group a and group b,the expression of CKIP-1 in group c was significantly higher(P<0.01),the expression of RUNX2 and ALPL was extremely low(P<0.01); compared with group d,the expression of CKIP-1 in group c was significantly lower(P<0.01),and the expression of RUNX2 and ALPL was significantly higher(P<0.01).The CKK-8 assay showed:compared with group a and group b,the level of cell proliferation in group c was significantly decreased(P<0.01)); compared with group c,the percentage of cell proliferation in group d after Qiangguyin treatment was increased,and the difference was significant(P<0.01).Alizarin red staining showed:compared with group a and group b,calcified nodules in group c decreased significantly; compared with group c,calcified nodules in group d increased significantly.Conclusion:The CKIP-1 overexpression is significantly inhibited the proliferation,osteogenic differentiation and mineralization of osteoblasts.Qiangguyin can increase osteoblast activity,osteogenic differentiation and mineralization in the pathological state of CKIP-1 overexpression ability.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(81373878)
△通信作者 E-mail:xlshi-2002@163.com
更新日期/Last Update: 2018-10-10