二硫代氨基吡咯烷对慢性混合反流性食管炎大鼠核因子-κB/白细胞介素-6信号通路的干预作用

Effects of pyrrolidine dithiocarbamate on nuclear factor-κB/interleukin-6 signaling pathway in rats with chronic mixed reflux esophagitis

目的 观察NF-κB抑制剂二硫代氨基吡咯烷（PDTC）对慢性混合反流性食管炎大鼠食管黏膜的保护作用及对NFκB/IL-6信号通路的影响.方法 40只健康清洁级SD雄性大鼠平均分为健康对照组、假手术组、模型对照组、奥美拉唑组、PDTC组,每组8只.除健康对照组和假手术组以外的大鼠建立慢性混合反流性食管炎大鼠模型.健康对照组、假手术组及模型对照组大鼠予以0.9％ NaCl溶液2 mL/d腹腔注射,奥美拉唑组大鼠每天按体质量予以20 mg/kg奥美拉唑腹腔注射,PDTC组大鼠每天按体质量予以100 mg/kg PDTC腹腔注射,6周后处死,肉眼及光学显微镜下观察食管组织的形态学改变并评分,应用ELISA法检测血清NF-κB p65、IL-6蛋白在各组大鼠中的表达.各组间均数比较采用t检验.结果 健康对照组、假手术组、模型对照组、奥美拉唑组、PDTC组5组大鼠的食管黏膜肉眼评分分别为0.000±0.000、0.000±0.000、2.250±0.707、1.125±0.835、1.429±0.535,病理评分分别为0.000±0.000、0.000±0.000、2.625±0.518、1.500±0.535、1.429±0.535,与模型对照组比较,健康对照组、假手术组、奥美拉唑组及PDTC组大鼠食管黏膜的肉眼、病理积分均降低,差异均有统计学意义（t肉眼 =7.603、7.603、2.909、2.506,t病理评分=9.674、9.674、4.277、4.399;P均＜0.05）.健康对照组、假手术组、奥美拉唑组、PDTC组大鼠的血清NF-κB p65蛋白含量分别为（68.618±18.450） pg/mL、（77.824±22.228） pg/mL、（106.693±45.312） pg/mL、（103.781±42.502） pg/mL,与模型对照组[（184.882±49.165） pg/mL]比较明显降低,差异均有统计学意义（t=6.262、5.612、3.308、3.427,P均＜0.05）;血清IL-6蛋白含量分别为（24.826±4.008） pg/mL、（23.599±4.351） pg/mL、（32.370±11.657） pg/mL、（33.694±10.394） pg/mL,与模型对照组[（51.378±9.697） pg/mL]比较亦显著降低,差异均有统计学意义（t=7.157、7.393、3.546、3.392,P均＜0.05）.PDTC组大鼠食管黏膜的肉眼、病理积分及血清NF-κB p65、IL-6蛋白含量与奥美拉唑组相比无明显升高,差异均无统计学意义（P均＞0.05）.结论 NF-κB抑制剂PDTC能减轻反流性食管炎大鼠食管黏膜的损伤程度,其机制可能与下调NF-κB/IL6信号通路有关.

Objective To observe the protective effects of nuclear factor （NF） κB inhibitor pyrrolidine dithiocarbamate （PDTC） on chronic mixed reflux esophagitis in rats and its influence on NF-κB/interleukin （IL）-6 signaling pathway.Method A total of 40 healthy male Sprague-Dawley （SD） rats were divided into healthy control group,sham operation group,model control group,omeprazole group and PDTC group with eight rats in each group.Except rats in healthy control group and sham operation group,mixed reflux esophagitis model were established in all the other groups.The rats of healthy control group,sham operation group and model control group were all intraperitoneally injected with 2 mL 0.9% NaCl,rats of omeprazole group were intraperitoneally injected with omeprazole 20 mg/kg,and rats of PDTC group were intraperitoneally injected with PDTC 100 mg/kg every day.After six weeks,the rats were sacrificed,the morphological changes of esophageal tissues were observed and scored by visual inspection and under light microscope.The serum levels of NF-κB p65 and IL-6 in rats of each group were assessed by enzyme linked immunoassay （ELISA）.t test was performed for mean comparison among groups.Results The scores of esophageal mucosa judged by visual inspection of healthy control group,sham operation group,model control group,omeprazole group and PDTC group were 0.000 20.000,0.000±0.000,2.250± 0.707,1.125 ± 0.835 and 1.429± 0.535,respectively.The pathological scores were 0.00020.000,0.000±0.000,2.625±0.518,1.500±0.535,1.429±0.535,respectively.Compared with those of model control group,the scores judged by visual inspection and the pathological scores of healthy control group,sham operation group,omeprazole group and PDTC group were lower,and the differences were statistically significant （t=7.603,7.603,2.909,2.506; t=9.674,9.674,4.277,4.399,all P〈0.05）.The serum levels of NF-κB p65 protein of healthy control group,sham operation group,omeprazole group and PDTC group were （68.618±18.450） pg/mL,（77.824±22.228） pg/mL,（106.693±45.312） pg/mL and （103.781± 42.502）pg/mL,respectively; compared with that of model group （（184.882±49.165） pg/mL）,which were significantly lower and the differences were statistically significant （t=6.262,5.612,3.308 and 3.427,all P〈0.05）.The serum levels of IL-6 protein were （24.826±4.008） pg/mL,（23.599±4.351） pg/mL,（32.370± 11.657） pg/mL and （33.694±10.394） pg/mL,respectively,which significantly decreased when compared with that of model group （（51.378±9.697） pg/mL,t=7.157,7.393,3.546 and 3.392,all P〈0.05）.There was no significant difference between PDTC group and omeprazole group in the score judged by visual inspection,pathological scores,the serum levels of NF-κB p65 and IL-6 protein （all P〉0.05）.Conclusion NF-κB inhibitor PDTC could reduce the injury severity of esophageal mucosal in reflux esophagitis rat,which mechanism might be related with the down-regulation of NF-κB/1L-6 signaling pathway.