摘要
目的:研究CEBPs各亚型在慢性肾损伤动物模型中的表达情况,探讨CEBPs在慢性肾脏疾病中可能作用机制,为阻断或延缓慢性肾损伤提供新的靶点。方法:建立阿霉素肾病(ADR)模型,制作各时间点肾组织切片及留取肾组织标本。利用免疫荧光染色和western blot检测CEBPs亚型(CEBP-β/CEBP-δ)在ADR肾病模型中表达情况。结果:(1)免疫荧光染色检测发现CEBP-β转录因子主要表达在肾小球及肾间质细胞核中,在ADR模型中阳性部位随着慢性肾损伤加重表达量逐渐下降。(2)western blot检测发现CEBP-β在ADR模型的4周和8周组较正常组相比表达量下降,且差异有统计学意义(P<0.05)。(3)western blot检测发现磷酸化的CEBP-β(p-CEBP-β)在ADR模型的4周和8周组较正常组相比表达量升高,与CEBP-β表达趋势相反。(4)western blot检测发现CEBP-δ在ADR模型4周组和8周组较正常组相比表达量升高,且差异具有统计学意义(P<0.05)。(5)NF-κBp65在ADR肾病模型4周组及8周组表达升高,较正常组比较差异有统计学意义(P<0.05)。且NF-κBp65表达趋势与CEBP-δ表达趋势相同,与CEBP-β表达趋势相反。结论:(1)CEBP-β/CEBP-δ可作为肾损伤的生物学标记物,参与慢性肾脏疾病的发生发展。(2)CEBP-β参与肾损伤机制可能与其磷酸化激活下游因子相关。(3)阻断CEBP-β/CEBP-δ转录因子的激活可能缓解肾脏炎症反应和纤维化反应的发生。(4)CEBP-β/CEBP-δ转录因子信号通路可能与NF-κB信号通路存在相互关联作用。
Objective：Study expression and mechanism of CEBP subtypes in chronic kidney disease model,for blocking or slowing the progression of chronic renal injury and providing a new target.Methods：Establishment of adriamycin （ADR） nephropathy model,making kidney tissue slices and kidney tissue specimens.Using immunofluorescence staining and western blot assay for the detection of CEBPs subtype （CEBP-β /p-CEBP-β/CEBP-δ） in adriamycin nephropathy model.Results：（1） Immunofluorescence staining detects CEBP-β transcription factor express in the renal glomerular and renal interstitial cell nucleus.The normal group of the ADR model showed the positive area.With chronic renal injurying,the expression of CEBP-β gradually decreased.（2） The results of western blot assay for detection of CEBP-β in ADR model shows expression in ADR 4 weeks and in ADR 8 weeks group is less than the normal group,and has significant difference （P0.05）.（3） While western blot assay detects the expression of p-CEBP-β in ADR 4 weeks and ADR 8 weeks model group is more than in the normal control group,and the trends contrary to expression of CEBP-β.（4） Western blot assay for detection of expression of CEBP-δ in ADR 4 weeks and 8 weeks model group more than in the normal control group,and has significant difference （P0.05）.（5） NF-κ B p65 in ADR 4 weeks and 8 weeks model group expression is elevated,compared with the normal group,and had significant difference （P0.05）.The trend of NF-κ B p65 is the same with the trend of CEBP-δ and opposite to the trend of CEBP-β.Conclusion：（1） CEBP-β /CEBP-δ can be used as a biological marker of renal injury,involving in occurrence and development of chronic kidney disease.（2） CEBP-β in renal injury mechanism is possibly that its phosphorylation activates the downstream factors.（3） Hypothesized that the blockade of CEBP-β /CEBP-δ transcription factor activation may relieve renal inflammation and fibrosis.（4） CEBP-β /CEBP-δ transcription factor
出处
《中国中西医结合肾病杂志》
2012年第11期963-966,I0004,共5页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
国家重点基础研究发展计划项目(No.2012CB517700)
国家自然科学基金资助项目(No.81270782
30771000)
上海市科学技术委员会科研计划重大项目(No.08dz1900502)
