CAJ | 학술논문

目的探讨克罗恩病(Crohn disease,CD)相关的NOD2及人β-防御素-2(human beta-defensin 2,hBD-2)基因多态性对hBD-2转录活性的影响及主要机理.方法将hBD-2报告基因质粒和NOD2真核表达载体共转染至HEK293T细胞,用脂多糖(LPS)和TNF-α分别孵育刺激后测定hBD-2转录活性变化.结果 LPS对hBD-2转录活性有抑制作用(P=0.020),TNF-α可相对升高hBD-2转录活性,呈剂量依赖性(P=0.004);UPS作用时,NOD2(P268S)改变前后hBD-2的转录活性差异有统计学意义(P=0.008);hBD-2(-233)改变前后hBD-2的转录活性差异无统计学意义(P=0.053).在TNF-α(5ng/ml)刺激时.NOD2(P268S)改变前后hBD-2的转录活性差异无统计学意义(P=0.064);hBD-2(-233)改变前后hBD-2的转录活性差异有统计学意义(P=0.006);NF-κB抑制剂可显著下调hBD-2转录的激活(P<0.001).结论 NOD2(P268S)改变能够降低hBD-2的内源性表达;hBD-2(233)改变导致hBD-2诱生性表达的减少;NF-κB通路可能是hBD-2诱导表达的主要路径.
목적 탐토극라은병(Crohn disease,CD)상관적NOD2급인β-방어소-2(human beta-defensin 2,hBD-2)기인다태성대hBD-2전록활성적영향급주요궤리.방법 장hBD-2보고기인질립화NOD2진핵표체재체공전염지HEK293T세포,용지다당(LPS)화TNF-α분별부육자격후측정hBD-2전록활성변화.결과 LPS대hBD-2전록활성유억제작용(P=0.020),TNF-α가상대승고hBD-2전록활성,정제량의뢰성(P=0.004);UPS작용시,NOD2(P268S)개변전후hBD-2적전록활성차이유통계학의의(P=0.008);hBD-2(-233)개변전후hBD-2적전록활성차이무통계학의의(P=0.053).재TNF-α(5ng/ml)자격시.NOD2(P268S)개변전후hBD-2적전록활성차이무통계학의의(P=0.064);hBD-2(-233)개변전후hBD-2적전록활성차이유통계학의의(P=0.006);NF-κB억제제가현저하조hBD-2전록적격활(P<0.001).결론 NOD2(P268S)개변능구강저hBD-2적내원성표체;hBD-2(233)개변도치hBD-2유생성표체적감소;NF-κB통로가능시hBD-2유도표체적주요로경.
Objective To explore the effects of polymorphisms of Crohn's disease related NOD2 gene and human beta-defensin 2 (hBD-2) on transcription of hBD-2 gene and its mechanism. Methods HEK293T cells were transfected with hBD-2 gene and NOD2 eukaryotic expression plasmid, and were then stimulated with LPS, TNF-α, or BAY 11-7082 (antagonist of NF-κB), respectively. Transcriptional activity of hBD-2 was detected afterwards. Results LPS could suppress transcription of hBD-2 (P=0. 020), which was increased by TNF-α in a dose-dependent manner (P =0. 004). In the presence of LPS, there was sig-nificant difference in transcriptional activity of hBD-2 between wild-NOD2 transfected group and mutated NOD2 (P268S) transfected group (P=0. 008), but there was no significant difference between wild hBD-2 transfected group and mutated hBD-2 transfected group (P=0. 053). With the stimulation of TNF-α (5 ng/ml), there was a significant difference between mutated hBD-2 transfected group and wild hBD-2 transfected group (P=0. 006), but no significant difference between wild-NOD2 transfected and mutated NOD2 transfected group was defected (P = 0. 064). Pretreatment with BAY 11-7082 before TNF-α (5 ng/ml) significantly inhibited the transcriptional activity of hBD-2 (P < 0. 001). Conclusion The poly-morphism of NOD2 affects the innate expression of hBD-2, the polymorphism of site in hBD-2 promoter (-233) may lead to significant decline of the inducible expression of hBD-2, and NF-κB might be a key pathway that NOD2 protein mediates the expression of defensin.