摘要
目的探讨细胞核转录因子-κB(NF-κB)在慢性氟中毒大鼠肾脏损伤机制中意义。方法原位末端转移酶标记法(TUNEL)及流式细胞学检测肾组织细胞凋亡率;免疫组化和实时荧光定量PCR(RT-PCR)检测肾组织p50、p65、IκBα和Bcl-2、Bax蛋白及mRNA表达。结果肾小管上皮细胞中p50和p65阳性表达强度分别为(134.65±9.71)、(113.62±10.49)、(105.30±8.00)和(119.74±5.17)、(111.90±6.90)、(101.71±9.31),核阳性细胞数随染氟增加而逐渐减少;IκBα表达[(152.67±3.36)、(155.75±3.37)、(156.29±3.78)]则呈逐渐增加趋势,差异均有统计学意义(P<0.05));与对照组比较,Bcl-2蛋白表达强度逐渐降低,Bax蛋白则逐渐升高(P<0.05);肾组织中mRNA表达与蛋白表达水平一致;凋亡细胞数随染氟增加逐渐增加(P<0.05);p65、p50及IκBα与Bcl-2/Bax比值呈相关性(r=0.696、0.720、-0.435,P<0.05)。结论 NF-κB相关基因表达减少引起的肾小管上皮细胞凋亡是慢性氟中毒大鼠肾脏病变的机制之一。
Objective To explore the significance of nuclear factor kappa B（NF-κB） related gene in the pathogenesis of the rats＇ kidney with chronic fluorosis.Methods Kidney cell apoptosis was detected with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling（TUNEL） and flow cytometry.Immunohistochemistry and real-time fluorescence quantitative PCR（RT-PCR） were used to detect the expressions of p50,p65,IκBα and Bcl-2,Bax protein and mRNA.Results The positive expressions of p50 and p65 in turbular epithelial cells were 134.65±9.71,113.62±10.49,105.30±8.00 and 119.74±5.17,111.90±6.90,101.71±9.31 for the rats in control,low fluorine,and high fluorine group,and the numbers of positive nuclear were decreased with the fluorine concerntration,but the expressions of IκBα were increased（152.67±3.36,155.75±3.37,156.29±3.78）,wtih significant differences.Compared with the control group,the expression of Bcl-2 was decreased and Bax＇s was increased（P0.05）.The expressions of all the indicatos＇ mRNA in the kidney were the same as those of the protein.The apoptosis cells were increased with the fluorine concentration（P0.05）.There were correlations between p50,p65,IκBα and the ratio of Bcl-2/Bax（r=0.696,r=0.720,r=-0.435,P0.05）.Conclusion The apoptosis of nephridial tubule epithelial cells caused by the reduction of NF-κB related gene is one of the pathogenesis in the kidney injury of chronic fluorosis in rats.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2011年第8期1021-1023,共3页
Chinese Journal of Public Health
基金
国家科技部国际合作项目(2010DFB30530)
贵阳市科技局科研基金([2009]筑科技合同字第3-007号)
关键词
NF-ΚB
氟中毒
肾小管上皮细胞
NF-κB
fluorine poisoning
nephridial tubule epithelial cell
