CAJ | 학술논문

目的观察脂多糖(LPS)作用下大鼠腹膜间皮细胞NF-κB活性及其对CD40和细胞间黏附分子1(ICAM-1)表达的影响.方法分离及培养大鼠腹膜间皮细胞.LPS不同浓度作用12 h 及LPS (5 μg/ml)作用不同时间点收集细胞;LPS(5 μg/ml)或BAY11-7085(一种IκBα的磷酸化抑制剂)不同浓度(1 μmol/L和5 μmol/L)预处理3 h加LPS,作用3 h后收集细胞;采用RT-PCR方法检测CD40和ICAM-1 mRNA表达.采用蛋白印迹检测NF-κB和磷酸化NF-κB(p-NF-κB)蛋白表达.结果与常规培养基对照组相比,5 μg/ml LPS组CD40和ICAM-1 mRNA表达显著升高(P<0.05),10 μg/ml LPS组显著高于5 μg/ml LPS作用组(P<0.05).5 μg/ml LPS 作用下,ICAM-1 mRNA表达从1 h开始升高,3 h达到高峰,之后逐渐降低.CD40 mRNA表达在1 h无显著变化,3 h时迅速达到高峰,之后逐渐降低.常规培养的大鼠腹膜间皮细胞结构性表达p-NF-κB蛋白;加入LPS后,p-NF-κB蛋白表达显著增加,其中30 min～1 h表达最强,之后逐渐降低,至2 h仍显著高于常规培养组(P<0.05).加入5 μmol/L BAY11-7085后,LPS诱导的CD40和ICAM-1 mRNA表达显著降低(P<0.05),与正常对照组相比差异无统计学意义. 结论 LPS以时间依赖和浓度依赖模式上调CD40和ICAM-1的表达.NF-κB信号途径参与调节LPS诱导的大鼠腹膜间皮细胞CD40和ICAM-1的表达.
목적 관찰지다당(LPS)작용하대서복막간피세포NF-κB활성급기대CD40화세포간점부분자1(ICAM-1)표체적영향.방법 분리급배양대서복막간피세포.LPS불동농도작용12 h 급LPS (5 μg/ml)작용불동시간점수집세포;LPS(5 μg/ml)혹BAY11-7085(일충IκBα적린산화억제제)불동농도(1 μmol/L화5 μmol/L)예처리3 h가LPS,작용3 h후수집세포;채용RT-PCR방법검측CD40화ICAM-1 mRNA표체.채용단백인적검측NF-κB화린산화NF-κB(p-NF-κB)단백표체.결과 여상규배양기대조조상비,5 μg/ml LPS조CD40화ICAM-1 mRNA표체현저승고(P<0.05),10 μg/ml LPS조현저고우5 μg/ml LPS작용조(P<0.05).5 μg/ml LPS 작용하,ICAM-1 mRNA표체종1 h개시승고,3 h체도고봉,지후축점강저.CD40 mRNA표체재1 h무현저변화,3 h시신속체도고봉,지후축점강저.상규배양적대서복막간피세포결구성표체p-NF-κB단백;가입LPS후,p-NF-κB단백표체현저증가,기중30 min～1 h표체최강,지후축점강저,지2 h잉현저고우상규배양조(P<0.05).가입5 μmol/L BAY11-7085후,LPS유도적CD40화ICAM-1 mRNA표체현저강저(P<0.05),여정상대조조상비차이무통계학의의. 결론 LPS이시간의뢰화농도의뢰모식상조CD40화ICAM-1적표체.NF-κB신호도경삼여조절LPS유도적대서복막간피세포CD40화ICAM-1적표체.
Objective To investigate the expression of CD40 and intercellular adhesion molecule-1 (ICAM-1) treated with lipopolysaccharide (LPS) in rat peritoneal mesothelial cells(RPMC) and the role of NF-κB signal transduction pathway. Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were exposed respectively to different concentrations of LPS for 12 h or treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of CD40 and ICAM-1, the RPMCs were treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of NF-κB and p-NF-κB protein, the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μmol/L or 1 μmol/L ) for 3 h, then treated with LPS for another 3 h, respectively. Expression of CD40 and ICAM-1 mRNA was examined by RT-PCR. Expression of NF-κB and p-NF-κB protein was detected by Western blot. Results Compared with medium control group, stimulation of RPMCs with 1 μg/ml and 5 μg/ml of LPS resulted in a significant increase in the expression of CD40 and ICAM-1 mRNA(P<0.05). 10 μg/ml of LPS had strongest effect on CD40 and ICAM-1 expression compared with that of 1 μg/ml and 5 μg/ml of LPS. Treatment with 5 μg/ml of LPS resulted in time-dependent increase in the gene level of CD40 and ICAM-1, with the peak at 3 h. However, after that time point, the gene level of them was gradually attenuated. Following treatment with LPS (5 μg/ml), the level of p-NF-κB began to increase at 15 min, gradually reached the peak at 1 h, and then decreased. But the level of p-NF-κB at 2 h was still significantly higher than that of medium control. 5 μmol/L of BAY11-7085 decreased significantly the up-regulation of CD40 and ICAM-1 induced by LPS. Conclusion LPS enhanced the expression of CD40 and ICAM-1 on RPMCs in a concentration-dependent and a time-dependent manner. LPS induced expression of CD40 and ICAM-1 depend on the NF-κB signal transduction pathway.