p38蛋白激酶对脂多糖诱导肺泡巨噬细胞NF-κB活化的调控  

Role of p38 Mitogen-activated Protein Kinase in the Activation of Nuclear Factor-κB in Lipopolysaccharide-stimulated Alveolar Macrophages

作　　者：黄翠萍[1] 杨和平[1] 张珍祥[2] 徐永健[2] 

机构地区：[1]湖北咸宁学院医学院内科,咸宁437100 [2]华中科技大学同济医学院附属同济医院呼吸内科,武汉430030

出　　处：《华中科技大学学报（医学版）》2007年第6期804-807,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的探讨p38蛋白激酶对脂多糖(LPS)诱导肺泡巨噬细胞NF-κB活化的调控机制。方法分离培养肺泡巨噬细胞。设正常对照组、LPS刺激组、SB203580(p38蛋白激酶抑制剂)+LPS组、PDTC(NF-κB抑制剂)+LPS组和SB203580+PDTC+LPS组。分别采用免疫细胞化学(SP法)和Western blot检测NF-κB、p38蛋白激酶和NF-κB抑制蛋白(I-κB)的表达。结果与正常对照组相比,LPS刺激肺泡巨噬细胞后胞核NF-κB和p38蛋白激酶染色显著增强,而I-κB的表达显著降低(均P<0.01)。SB203580、PDTC均可显著降低LPS刺激的肺泡巨噬细胞NF-κB,p38蛋白激酶胞核染色阳性细胞的百分比,并显著增强I-κB的表达(均P<0.05)。同时应用PDTC和SB203580预孵育肺泡巨噬细胞,与LPS刺激组相比,p38蛋白激酶和NF-κB胞核染色阳性细胞百分比均显著降低(P<0.05),I-κB的表达显著增强(P<0.05),但分别与SB203580+LPS和PDTC+LPS组相比,差异均无显著性意义(均P>0.05)。结论LPS刺激肺泡巨噬细胞p38蛋白激酶和NF-κB活化;p38蛋白激酶可能通过调节I-κB降解而调控NF-κB的活化,NF-κB可能是p38信号途径下游的最重要位点。Objective To explore the role of p38 mitogen-activated protein kinase （MAPK） in the activation of nuclear fac- tor-κB （NF-κB） in lipopolysaccharide （LPS）-stimulated alveolar macrophages （AMs）. Methods AMs were isolated and purified from normal rats and divided into 5 groups： normal group, LPS-stimulated group, SB203580＋LPS group, PDTC＋LPS group and SB203580＋PDTC＋LPS group. By using immuocytochemical staining and Western blot, the expression of NF-κB, p38 protein and I-κB was detected. Results In LPS-stimulated AMs, the percentage of positive cells of active NF-κB and p38 protein was higher than that in normal AMs （P〈0.01）, which was increased from （7. 34±1.33）%, （6. 12±2. 15）% to （38.52±9.71） %, （35.20±8.51）%, and the expression of I-κB was decreased significantly [（0. 3012±0. 0431） vs （0. 7615± 0. 0924）, P〈0.01]. Either SB203580 or PDTC could decrease the percentage of positive cells of active NF-κB and p38 protein and increase the expression of I-κB （P〈0.05）. In the AMs cultured with both SB203580 and PDTC simultaneously, the percentage of positive cells of active NF-κB and p38 protein was decreased and the expression of I-κB increased as compared with those in LPS-stimulated AMs （P〈0.05）, but there was no significant difference in comparison to those in SB203580q-LPS group or PDTC＋LPS group （P〉0.05）. Conclusion LPS can stimulate the activation of p38 protein and NF-κB in AMs and p38 protein can regulate the activation of NF-κB by regulating the degradation of I-κB. NF-κB may be an important target in p38 protein signal pathway.

关 键 词：P38蛋白激酶 核因子-ΚB 脂多糖 肺泡巨噬细胞 

分 类 号：Q555.7[生物学—生物化学]