Shh信号通路对髁突肥大软骨细胞凋亡活性影响的研究
Effects of Shh signaling pathway on apoptotic activity of the chondrocytes in the cartilage with condylar hyperplasia
张武阳 1陈宇翔 1杨璇璇 2贾骏麒 1张玉灿 1房维 3刘洋 1常士平 1薄斌 1马秦 1龙星3
作者信息
- 1. 军事口腔医学国家重点实验室,口腔疾病国家临床医学研究中心,陕西省口腔疾病临床医学研究中心,第四军医大学口腔医院:颌面外科,陕西 西安 710032
- 2. 军事口腔医学国家重点实验室,口腔疾病国家临床医学研究中心,陕西省口腔疾病临床医学研究中心,第四军医大学口腔医院:正畸科,陕西 西安 710032
- 3. 口腔生物医学教育部重点实验室,武汉大学口腔医院颌面外科,湖北 武汉 430079
- 折叠
摘要
目的:探讨Sonic hedgehog(Shh)信号通路对髁突肥大软骨细胞凋亡活性的影响.方法:取正常髁突软骨3例,髁突肥大软骨6例,酶消化法分离培养其软骨细胞后,用Western blotting和Real-time PCR法分别检测两种软骨细胞中凋亡效应因子cleaved-caspase-3和BCL-2的表达;用免疫组化和Western blotting法比较正常髁突软骨和髁突肥大软骨中Shh和Smo的表达;用Shh、Cyclopamine、LY294002和U0126分别刺激两种软骨细胞,观察cleaved-caspase-3和BCL-2表达的变化.结果:与正常髁突软骨相比,髁突肥大软骨细胞中cleaved-caspase-3的表达水平降低,BCL-2 mRNA的表达水平升高(P<0.05);在髁突肥大软骨中Shh、Smo呈高表达;Shh刺激组软骨细胞的凋亡活性降低(P<0.05),Cyclopamine刺激组软骨细胞的凋亡活性增加(P<0.05),LY294002及U0126刺激组软骨细胞的BCL-2 mRNA表达水平均降低(P<0.05).结论:Shh可通过介导PI3K/AKT和MAPK/ERK通路而抑制髁突肥大软骨细胞的凋亡,促进髁突肥大的病理进程.
Abstract
AIM:To investigate the effect of Shh signaling pathway on the apoptosis of the chondrocytes in the cartilage with condylar hyperplasia(CH).METHODS:Chondrocytes were isolated from 6 CH and 3 normal cartilage specimens.Western blotting and Real-timePCR were used to detect the expression levels of cleaved-caspase-3 and BCL-2.Then the expression levels of Shh and Smo were assessed by immunohistochemistry and Western blotting.Finally,chondrocytes were treated with Shh,cyclopamine,LY294002 or U0126,the expression levels of cleaved-caspase-3 and BCL-2 were examined.RESULTS:As compared with normal chondrocytes,CH chondrocytes showed lower apoptotic activity and expressed higher level of BCL-2 mRNA and lower level of cleaved-caspase-3 (P<0.05).Shh and Smo had higher expression in CH chondrocytes.Shh stimulation significantly inhibited the apoptosis of chondrocytes.On the contrary,cyclopamine stimulation significantly increased the apoptosis of chondrocytes.LY294002 and U0126 treatments reduced BCL-2 mRNA expression in both groups (P<0.05).CONCLUSION:Shh promotes human TMJ cartilage overgrowth in the developing process of CH by inhibiting chondrocyte apoptosis via PI3K/AKT and MAPK/ERK pathway.
关键词
髁突肥大/Sonichedgehog/凋亡Key words
condylar hyperplasia/sonic hedgehog/apoptosis引用本文复制引用
出版年
2017
NETL
NSTL
维普
万方数据