摘要
目的探讨基质金属蛋白酶3(MMP3)基因与性别决定相关高迁移率簇蛋白盒基因9(Sox9)调控在人椎间盘退变髓核细胞中的作用。方法构建MMP3与Sox9慢病毒表达载体,感染人退变髓核细胞,按照处理方式不同分为4组,分别为空白对照组(A组)、MMP3沉默转染组(B组)、Sox9过表达组(C组)以及MMP3沉默+Sox9过表达共转组(D组),各组髓核细胞置于恒温培养箱中培养,定期换液。72h后,通过荧光定量PCR方法检测转染人退变髓核细胞Ⅱ型胶原、蛋白多糖mRNA的表达,通过Western Blot方法检测转染人退变髓核细胞Ⅱ型胶原、蛋白多糖蛋白的表达。结果 B、C、D组蛋白多糖、Ⅱ型胶原蛋白及mRNA表达水平与A组比较明显增高,差异具有显著性(F=84.50~413.09,q=4.57~49.13,P<0.01);D组蛋白多糖、Ⅱ型胶原蛋白及mRNA表达水平与B、C组比较,差异具有显著性(q=13.72~30.24,P<0.01)。结论 MMP3基因沉默、Sox9基因过表达可以促进人退变髓核细胞蛋白多糖、Ⅱ型胶原的分泌,延缓椎间盘退变,且二者之间具有协同作用。
Objective To investigate the regulatory effect of matrix metalloproteinase 3（MMP3）gene and Sox9 gene on human degenerative nucleus pulposus cells. Methods The lentiviral expression vectors of the MMP3 and Sox9 was constructed and transfected into human degenerative nucleus pulposus cells.According to the treatment method,the cells were divided into blank control group（group A）,MMP3 silencing group（group B）,Sox9 overexpression group（group C）,and MMP3 silencing＋Sox9 overexpression group（group D）.All groups of nucleus pulposus cells were placed and cultured in a constant-temperature incubator and the medium was changed regularly.After 72 hours,qualitative real-time PCR was used to measure the mRNA expression of typeⅡ collagen and proteoglycan in transfected human degenerative nucleus pulposus cells,and Western blot was used to mea-sure the protein expression of typeⅡcollagen and proteoglycan in these cells. Results Groups B,C,and D had significantly higher protein and mRNA expression of proteoglycan and typeⅡ collagen than group A（F=84.50-413.09,q=4.57-49.13,P0.01）.There were significant differences in the protein and mRNA expression of proteoglycan and type Ⅱ collagen between group D and groups B and C（q=13.72-30.24,P0.01）. Conclusion MMP3 silencing and Sox9 overexpression can promote the secretion of proteoglycan and typeⅡ collagen in human degenerative nucleus pulposus cells and thus delay intervertebral disc dege-neration,and there is a synergistic effect between the two genes.
出处
《青岛大学医学院学报》
CAS
2017年第4期386-389,共4页
Acta Academiae Medicinae Qingdao Universitatis
基金
国家自然科学基金资助项目(81470104)
