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目的探讨核转录因子κB(NF-κB)及其诱导的细胞因子在大鼠角膜炎中的表达及二硫代氨基甲酸吡咯烷(PDTC)对其表达的影响。方法选择112只大鼠建立角膜炎模型,按随机数字表法分为炎性组56只和PDTC预处理组56只．模型制作前30 min,大鼠球结膜下分别注射生理盐水(炎性组)和PDTC(PDTC预处理组),每组按脂多糖(LPS)刺激后不同时间又分为0.5、1．0、3．0、6．0、12．0、24.0及72．0 h亚组,每亚组8只鼠。裂隙灯显微镜下观察大鼠的眼部变化;病理组织切片观察角膜形态学改变;免疫组织化学染色检测角膜NF-κB p65的表达;逆转录聚合酶链反应(RT- PCR)检测角膜肿瘤坏死因子α(TNF-α)mRNA的表达。结果LPS刺激后炎性组大鼠角膜组织明显水肿,大量炎性细胞浸润,胶原纤维排列紊乱;免疫组织化学染色显示LPS刺激后0．5 h即可见NF-κB阳性细胞,并表达逐渐增强,3．0～12．0 h最强,0.5～24．0 h间均较PDTC预处理组明显增多(P< 0．01);TNF-αmRNA的表达在LPS刺激后0．5 h就开始升高,3.0～12．0 h最强,0．5～24．0 h间均较PDTC预处理组明显增高(P<0．01)。结论NF-κB及其诱导的TNF-α在角膜炎中发挥重要作用, PDTC可能通过抑制NF-κB的活性减轻角膜损伤。
Objective To investigate the expression of nuclear factor kappa B (NF-κB) and its induced cytokines in rat keratitis and the effect of pyrrolidine dithiocarbamate (PDTC) on its expression. Methods 112 rats were selected to establish keratitis model. According to the random number table, 56 patients were divided into inflammatory group and PDTC pretreatment group. Rats in each group were injected subcutaneously with saline (inflammatory group) and PDTC (pretreatment group) respectively 30 minutes before the model was made. Each group was divided into 0.5, 1.0, 3.0, 6.0, 12.0, 24.0 and 72.0 h subgroups, 8 mice per subgroup. The changes of corneal morphology were observed under slit lamp microscope. The corneal morphology was observed by histological sections. The expression of NF-κB p65 was detected by immunohistochemical staining. The expression of corneal tumor necrosis factor-α (TNF-α) was detected by reverse transcription-polymerase chain reaction α (TNF-α) mRNA expression. Results The corneal edema, inflammatory cell infiltration and collagenous fibers were disordered in LPS-stimulated rats. Immunohistochemical staining showed that NF-κB positive cells were observed at 0.5 h after LPS stimulation, 3.0 ~ 12.0 h was the strongest, 0.5 ~ 24.0 h were significantly higher than the PDTC pretreatment group (P <0.01); TNF-αmRNA expression began 0.5 h after LPS stimulation High, 3.0 ~ 12.0 h strongest, 0.5 ~ 24.0 h were significantly higher than PDTC pretreatment group (P <0.01). Conclusion NF-κB and its induced TNF-α play an important role in keratitis. PDTC may reduce corneal injury by inhibiting the activity of NF-κB.