Notch信号通路对骨关节炎软骨细胞增殖和软骨基质合成的作用研究  

Effects of Notch Signaling Pathway on Chondrocyte Proliferation and Cartilage Matrix Synthesis Osteoarthritis

作　　者：林飞太[1,2] 林煜[1] 林钡 陈赛楠[3] 翁艳[1] 冯尔宥[1,2] LIN Fei-tai;LIN Yu;LIN Bei;CHEN Sai-nan;WENG Yan;FENG Er-you(Department of Arthrosis Surgery,Fuzhou Second Hospital,Fuzhou 350007,China;The Third Clinical Medical College,Fujian Medical University,Fuzhou 350122,China;Fujian Institute of Traditional Chinese Medicine,Fuzhou 350003,China)

机构地区：[1]厦门大学附属福州市第二医院关节外科,福州350007 [2]福建医科大学第三临床医学院,福州350122 [3]福建中医药研究院,福州350003

出　　处：《中国合理用药探索》2023年第2期110-118,共9页Chinese Journal of Rational Drug Use

基　　金：福州市科技项目(2020-WS-118);福建省骨科创伤急救与康复临床医药研究中心(2020Y2014)。

摘　　要：目的:探索Notch信号通路对白介素-1β(IL-1β)诱导软骨细胞增殖和软骨基质合成的调控作用。方法:培养C28/I2软骨细胞系,用10ng/ml IL-1β诱导凋亡,采用Ad-NICD感染过表达NICD和si-NICD转染沉默NICD,Real-time PCR和Western Blot检测过表达和沉默效率,CCK-8检测细胞增殖,检测PCNA、ColⅡ和ACAN mRNA和蛋白的表达水平。结果:Ad-NICD的病毒感染复数(MOI)为40时过表达效果最佳,si-NICD的沉默效率最佳;Ad-NICD感染C28/I2可抑制细胞增殖和抑制PCNA、ColⅡ和ACAN mRNA和蛋白的表达水平(P<0.05);si-NICD3转染C28/I2可促进细胞增殖和促进PCNA、ColⅡ和ACAN mRNA和蛋白的表达水平(P<0.05)。结论:过表达NICD激活Notch通路可抑制IL-1β诱导C28/I2软骨细胞增殖和软骨基质合成,沉默NICD阻遏Notch通路可促进细胞增殖和软骨基质合成。Objective:To investigate the regulation of Notch signaling pathway on chondrocyte proliferation and cartilage matrix synthesis induced by interleukin-1β(IL-1β).Methods:C28/I2 chondrocyte line was cultured,and 10ng/ml IL-1βwas used to induce apoptosis.Overexpressed NICD was infected with Ad-NICD and its expression was silenced by si-NICD.Real-time PCR and Western Blot were used to detect overexpression and silencing efficiency.Cell proliferation was detected by CCK-8.The mRNA and protein expression levels of PCNA,colⅡand ACAN were detected.Results:The overexpression effect of multiplicity of infection(MOI)40 was the best,and the silencing efficiency of si-NICD was the best.Ad-NICD infection with C28/I2 inhibited cell proliferation and the mRNA and protein expression levels of PCNA,ColⅡand ACAN(P<0.05).si-NICD3 transfection with C28/I2 can promote cell proliferation and the mRNA and protein expression levels of PCNA,ColⅡand ACAN(P<0.05).Conclusion:Overexpression of NICD in C28/I2 can inhibit cell proliferation and cartilage matrix synthesis,and silencing NICD can promote cell proliferation and cartilage matrix synthesis.

关 键 词：骨关节炎 软骨细胞 NOTCH信号通路 NICD 细胞外基质 

分 类 号：R246.9[医药卫生—针灸推拿学]