摘要
目的小干扰RNA技术沉默结缔组织生长因子(CTGF)表达,并观察对类风湿关节炎(RA)成纤维样滑膜细胞(FLS)凋亡及对部分凋亡相关基因的影响。方法脂质体转染CTGF siRNA至FLS,real time PCR筛选出有效的CTGF siRNA。使FLSCTGF表达沉默后,采用FITC-PI双染法通过流式细胞仪检测对FLS凋亡的影响。Real-time PCR检测CTGF基因沉默对bax、bcl-xl和survivin等凋亡相关基因表达的影响。结果筛选出H-1序列是针对人CTGF的特异性SiRNA,抑制率>70%。在外源性CTGF作用下,由去血清导致的FLS凋亡明显减少;沉默FLS中的CTGF基因表达,FLS的凋亡明显增加。在FLS中CTGF基因表达沉默后,Bax基因表达上调,bcl-xl基因表达下调,但差异没有统计学意义(P>0.05)。survivin基因表达出现明显下调(P<0.01)。结论 CTGF减少RA FLS凋亡的保护性作用很可能是通过维持survivin表达来实现的。
Objective To silence the expression of CTGF by small interfering RNA technology, to observe the influence on fibroblast-hke synovial cell apoptosis and several apoptosis-related genes, and to explore the mechanism of action of CTGF in rheumatoid arthritis synovial lesions. Methods Effective CTGF siRNA was screened through real-time PCR. The influence of CTGF siRNA on FLS apoptosis was detected with FITC-PI double staining by flow cytometry, bax, bcl-xl and survivin were detected using real-time PCR when CTGFmRNA has been silenced. Results Compared with other 2 groups of ohgo and NC ohgo, H 1 oligo exhibited the strongest interfering action to CTGF （ inhibition ratio〉70% ）, so that it is selected as the effective target gene sequence for the following experiment. Apoptosis of FLS induced by serum deprivation was significantly decreased in the presence of exogenous CTGF. When expression of the CTGF gene was knocked down in FLS, FLS apoptosis was significantly increased, and expres- sion levels of surivin mRNA were decreased significantly （P 〈 0.01 ）. Conclusion FLS survival is positively regulated by CTGF, which may through the sustaining the expression of survivin.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2016年第5期430-433,共4页
Journal of China Medical University
基金
辽宁省医学高峰建设工程专项经费(2010006)
关键词
结缔组织生长因子
成纤维样滑膜细胞
凋亡
connective tissue growth factor
fibroblast-like synoviocytes
apoptosis
