环境内分泌干扰物双酚A对非酒精性脂肪性肝病大鼠模型肠道黏膜屏障的影响  

Influence of bisphenol A on intestinal mucosal barrier in rats with nonalcoholic fatty liver disease

作　　者：丁雯瑾[1] 袁涛[2] 沈峰[1] 周达[1] 孙超[1] 范建高[1] 陈源文[1] DING Wenfin;YUAN Tao;SHEN Feng(Department of Gastroenterology,Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China)

机构地区：[1]上海交通大学医学院附属新华医院消化科,上海200092 [2]上海交通大学环境科学与工程学院,上海200240

出　　处：《临床肝胆病杂志》2018年第6期1268-1272,共5页Journal of Clinical Hepatology

基　　金：国家自然科学基金资助项目(81400610);教育部留学回国人员科研启动基金项目(201404802);上海市卫生和计划生育委员会青年科研项目(20134Y043);上海交通大学医工交叉研究基金资助项目(YG2016MS72);黎介寿院士肠道屏障研究专项基金(LJS_201214;LJS_201319)

摘　　要：目的研究环境内分泌干扰物双酚A(BPA)对非酒精性脂肪性肝病(NAFLD)大鼠炎症反应的影响,以及对肠道黏膜屏障的损害作用。方法将18只成熟雄性SD大鼠随机分为3组:正常组、NAFLD组和NAFLD+BPA组,每组6只。光镜下观察肝脏病理改变,ELISA检测血清TNFα、IL-1β、IL-6和IL-8等炎症因子,鲎试剂终点比色法检测内毒素水平,免疫荧光法观察肠道黏膜Occludin蛋白的表达情况,及实时聚合酶链式反应测定肠道黏膜外周蛋白ZO-1 mRNA的改变。计量资料多组间比较采用单因素方差分析,进一步两两比较用SNK-q检验。结果肝脏病理学证实NAFLD模型建模成功。100 nmol/L BPA摄入8周后,TNFα、IL-6、IL-8炎症因子表达上调[分别为(127.65±22.40)pg/ml、(199.34±17.46)pg/ml和(258.79±12.82)pg/ml]伴内毒素水平增高[(0.88±0.26)EU/ml],与正常组[分别为(64.87±10.83)pg/ml、(91.27±9.82)pg/ml、(123.76±19.68)pg/ml和(0.27±0.09)EU/ml]及NAFLD组[分别为(92.34±10.68)pg/ml、(181.93±20.11)pg/ml、(201.64±22.34)pg/ml和(0.63±0.15)EU/ml]相比,差异均有统计学意义(P值均<0.05)。NAFLD组和NAFLD+BPA组中的IL-1β水平明显高于正常组[(186.31±20.06)pg/ml、(208.78±13.77)pg/ml)vs(112.84±23.12)pg/ml,P值均<0.05],但NAFLD组和NAFLD+BPA组间差异无统计学意义。此外,与正常组比较,NAFLD组和NAFLD+BPA组中肠道紧密连接蛋白Occludin水平下降,肠道黏膜外周蛋白ZO-1 mRNA表达水平呈显著降低,BPA干预促进ZO-1 mRNA进一步下调(68.03±11.73、45.24±6.98、33.25±11.04,两两组间比较,P值均<0.05)。结论在NAFLD背景下,长期低剂量BPA的暴露能加重炎症因子释放,促进内毒素血症,进而损伤肠道黏膜屏障。Objective To investigate the influence of bisphenol A（BPA）,an environmental endocrine disruptor,on inflammatory response in rats with nonalcoholic fatty liver disease（NAFLD）,as well as its adverse effect on intestinal mucosal barrier. Methods A total of 18 mature male rats were randomly divided into normal group,NAFLD group,and NAFLD ＋ BPA group,with 6 rats in each group. liver pathological changes were observed under a light microscope; ELISA was used to measure the serum levels of tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β）,interleukin-6（IL-6）,and interleukin-8（IL-8）; the terminal colorimetric analysis with Limulus amebocyte lysate was used to measure the level of endotoxin; an immunofluorescence assay was used to measure the expression of occluding protein in the intestinal mucosa; real-time PCR was used to measure the mRNA expression of ZO-1 in the intestinal mucosa. A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the SNK-q test was used for further comparison between two groups. Results Liver pathological examination showed that the rat model of NAFLD was established successfully. After the 8-week treatment with 100 nmol/L BPA,compared with the normal group and the NAFLD group,the NAFLD ＋ BPA group had significant increases in the expression of TNF-α（127. 65 ± 22. 4 pg/ml vs 64. 87 ± 10. 83 pg/ml and 92. 34 ± 10. 68 pg/ml,P 0. 05）,IL-6（199. 34 ± 17. 46 pg/ml vs 91. 27 ± 9. 82 pg/ml and 181. 93 ± 20. 11 pg/ml,P 0. 05）,and IL-8（258. 79 ± 12. 82 pg/ml vs 123. 76 ±19. 68 pg/ml and 201. 64 ± 22. 34 pg/ml,P 0. 05） and the level of endotoxin（0. 88 ± 0. 26 EU/ml vs 0. 27 ± 0. 09 EU/ml and 0. 63 ±0. 15 EU/ml,P 0. 05）. The NAFLD group and the NAFLD ＋ BPA group had a significantly higher level of IL-1β than the normal group（186. 31 ± 20. 06 pg/ml and 208. 78 ± 13. 77 pg/ml vs 112. 84 ± 23. 12 pg/ml,both P 0. 05）,but there was no significant difference between these two groups. Compared w

关 键 词：非酒精性脂肪性肝病 内分泌干扰物 肠黏膜 大鼠 SPRAGUE-DAWLEY 

分 类 号：R575.5[医药卫生—消化系统]