摘要
目的研究氧化应激对猪眼小梁细胞NF-κB信号通路相关蛋白P65及MMPs/TIMPs表达的影响,探讨氧化应激介导的NF-κB信号通路在猪小梁细胞外基质重塑中的作用。方法传第三代的猪眼小梁细胞血清饥饿培养24 h后,利用NF-κB的特异抑制剂吡咯二硫氨基甲酸(pyrrolidine dithocarbamate,PDTC)进行预处理,或直接用H2 O2进行干预,细胞免疫组化和RT-PCR检测猪眼小梁细胞NF-κB P65、M M P-2、M M P-3和TIM P-1的表达。结果 H2O2处理组猪眼小梁细胞NF-κB P65、MMP-2、MMP-3和TIMP-1的表达明显增加,与对照组比较,差异均有统计学意义。PDTC组H2O2诱导的猪眼小梁细胞MMP-2、MMP-3的表达,但仍高于对照组猪眼小梁细胞的表达。PDTC对H2O2诱导的猪眼小梁细胞TIMP-1的表达无影响。结论氧化应激可能通过诱导猪眼小梁细胞NF-κB信号通路的激活,参与猪小梁细胞外基质的重塑,可能还有其他的信号通路参与这一过程。
Objective To investigate the effects of oxidative stress on expressions of NF-κB P65,MMPs/TIMPs in porcine trabecular meshwork cells（PTCs） and to explore the role of the NF-κB signaling pathway in trabecular extracelluar matrix（ECM） remodeling.Methods The PTCs were cultured and passaged.The 3rd generation trabecular cells were cultured without serum and stimulated by 1 mmol/L H2O2 with or without pretreatment of pyrrolidine dithiocarbamate（PDTC）.Expressions of NF-κB P65,MMP-2,MMP-3 and TIMP-1 were analyzed with immunocytochemistry and RT-PCR.The data were statistically analyzed.Results Expressions of NF-κB P65,MMP-2,MMP-3 and TIMP-1 were significantly increased by H2O2 in PTCs.PDTC partly reversed H2O2 induced expressions of MMP-2 and MMP-3.Expression of TIMP-1 had no significant variation after pre-treatment of PDTC.Conclusion Oxidative stress can induce expression of NF-κB P65 in PTCs.The NF-κB signaling pathway was involved in trabecular ECM remodeling.Other signaling pathways may be related to the process.
出处
《山东大学学报（医学版）》
CAS
北大核心
2012年第5期51-54,59,共5页
Journal of Shandong University:Health Sciences
基金
山东省中青年科学家科研奖励基金资助(2009BSB01638)
山东省卫生厅科技计划资助项目(2009HZ105)
山东省自然科学基金资助(Y2008C127)
山东省教育厅科技计划资助项目(J08LG13)
关键词
氧化应激
小梁细胞
NF-ΚB信号通路
吡咯二硫代氨基甲酸
细胞外基质
重塑
Oxidative stress
Trabecular meshwork cell
NF-κB signaling pathway
Pyrrolidine dithiocarbamate
Extracelluar matrix
Remodelling
