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目的探讨 NF-kB 及其诱导 TNFα在大鼠铜绿假单胞菌(PA)肺炎中的表达及二硫代氨基甲酸吡咯烷(PDTC)对其的影响。方法 72只 SD 大鼠分为健康对照组、PA 组、PDTC 干预组,每组24只。PA 组大鼠制作 PA 模型,PDTC 干预组制作 PA 模型前腹腔注射 PDTC 200 mg/kg 体重,PA组、PDTC 干预组接种铜绿假单胞菌悬液0.2ml(6×10~8CFU/ml)。观察大鼠肺组织形态学改变,免疫组化及 Western blot 检测肺组织 NF-kB 表达,RT-PCR 检测肺组织 TNFαmRNA 表达。结果 PA 组大鼠肺组织明显充血、水肿,大量炎性细胞浸润;PDTC 干预组大鼠肺组织充血、水肿等炎性表现较 PA组明显减轻。PA 组大鼠肺组织 NF-kB 表达阳性细胞明显增多,PDTC 干预组 NF-kB 表达阳性细胞明显减少。Western blot 显示,各时间点 NF-kB 表达不同,3h 达高峰。RT-PCR 显示,PA 组 TNFαmRNA高表达,以6h 最为明显;PDTC 干预组 TNFαmRNA 表达明显下调,与 PA 组相比,差异有统计学意义(P<0.01)。结论 NF-kB 及其诱导的 TNFα在 PA 肺炎中发挥重要作用,PDTC 可能通过抑制NF-kB 的活性减轻 PA 引起的肺损伤。
Objective To investigate the expression of NF-κB and its inducing TNFα in Pseudomonas aeruginosa (PA) pneumonia and the effect of PDTC on it. Methods 72 SD rats were divided into healthy control group, PA group and PDTC intervention group, 24 rats in each group. The PA model was made in PA group. The PDTC intervention group was given intraperitoneal injection of 200 mg / kg body weight of PDTC. PA and PDTC groups were inoculated with 0.2 ml (6 × 10 ~ 8 CFU / ml) of Pseudomonas aeruginosa suspension. The morphological changes of lung tissue were observed. The expression of NF-κB in lung tissue was detected by immunohistochemistry and Western blot. The expression of TNFα mRNA in lung tissue was detected by RT-PCR. Results The pulmonary tissue of PA rats was hyperemic, edematous and a large number of inflammatory cells were infiltrated. Compared with PA group, inflammatory reaction such as congestion and edema in lung tissue of rats in PDTC intervention group was significantly reduced. The expression of NF-κB positive cells in PA group was significantly increased, while the expression of NF-κB positive cells in PDTC group was significantly decreased. Western blot showed that the expression of NF-kB at different time points reached the peak at 3h. RT-PCR showed that TNFαmRNA was highly expressed in PA group, which was the most obvious at 6h; the expression of TNFαmRNA in PDTC group was significantly down-regulated compared with PA group (P <0.01). Conclusion NF-κB and TNF-α induced by it play an important role in PA pneumonia. PDTC may reduce PA-induced lung injury by inhibiting the activity of NF-κB.