LPS致炎时间对小鼠小肠组织中IAP与NF-κB表达的影响

doi:10.16431/j.cnki.1671-7236.2023.11.011

摘要/Abstract

摘要： 【目的】探究随着脂多糖（LPS）致炎时间延长小鼠小肠组织中肠型碱性磷酸酶（IAP）和核转录因子-κB（NF-κB）表达量变化，为减少肠黏膜应激损伤提供理论基础。【方法】选用体重相近、日龄接近的雄性健康ICR小鼠35只，随机分为7组，每组5只，小鼠腹腔注射1 mL/只LPS（5 mg/kg）。分别在注射LPS前（0 h）与注射后3、6、12、24、48和72 h，各组小鼠麻醉后断颈处死，采集十二指肠、空肠、回肠组织。用实时荧光定量PCR及Western blotting分别检测各肠段中IAP与NF-κB的mRNA水平及蛋白表达量；通过免疫共沉淀法检测IAP与NF-κB之间相互作用。【结果】与0 h相比，随着LPS致炎时间延长小鼠小肠组织中IAP基因表达量呈现波浪式变化，十二指肠、空肠组织在6 h时达到最高（P<0.01），回肠组织在48 h达到最高（P<0.01）；24 h时十二指肠、空肠组织中NF-κB基因表达量极显著或显著升高（P<0.01；P<0.05），其他时间点表达量变化均差异不显著（P>0.05）。LPS处理后，随着时间延长，小鼠各肠段中IAP和NF-κB蛋白表达量均呈现先升高后降低的趋势，且在6 h时蛋白表达量达到最高（P<0.05）。免疫共沉淀结果显示，NF-κB与IAP存在相互作用。【结论】LPS致炎过程中，NF-κB和 IAP的mRNA和蛋白表达量随着肠道炎症发生而升高，在致炎后 6 h表达量达到最高，但随着致炎时间延长表达量逐渐降低，在肠黏膜应激损伤过程中IAP和NF-κB存在相互协同作用。

关键词: 脂多糖; 肠黏膜损伤; IAP; NF-κB

Abstract: 【Objective】 The purpose of this experiment was to investigate the changes of the expression levels of intestinal alkaline phosphatase (IAP) and nuclear transcription factor-κB (NF-κB) in mice with the prolonged inflammatory time induced by lipopolysaccharide (LPS), and provide a theoretical basis for reducing intestinal mucosal stress damage.【Method】 A total of 35 healthy male ICR mice with similar body weight and age were randomly divided into 7 groups with 5 mice in each group.The mice were intraperitoneally injected with 1 mL/mouse LPS (5 mg/kg).Before (0 h) and after injection (3, 6, 12, 24, 48 and 72 h) of LPS, the mice in each group were killed by neck dissection after anesthesia, and duodenum, jejunum and ileum tissues were collected.Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression of IAP and NF-κB in each intestinal segment.The interaction between IAP and NF-κB was detected by co-immunoprecipitation.【Result】 Compared with 0 h, IAP gene expression in the small intestine of mice showed a wave change with the prolongation of LPS inflammatory time, the highest level was reached in duodenum and jejunum at 6 h (P < 0.01), and the highest level was reached in ileum at 48 h (P < 0.01).At 24 h, the expression of NF-κB gene in duodenum and jejunum were extremely significantly or significantly increased (P <0.01 or P < 0.05), there were no significant differences in expression levels at other time points (P > 0.05).After LPS treatment, the expression of IAP and NF-κB protein in each intestinal segment of mice showed a trend of first increasing and then decreasing, and the protein expression reached the highest level at 6 h (P < 0.05).The results of co-immunoprecipitation showed that NF-κB interacted with IAP.【Conclusion】 In the inflammatory process of LPS, the mRNA and protein expressions of NF-κB and IAP increased with the occurrence of intestinal inflammation, and reached the highest level at 6 h after inflammation, but gradually decreased with the extension of inflammatory time.There was a synergistic effect between IAP and NF-κB in the process of intestinal mucosal stress injury.

Key words: lipopolysaccharide; intestinal mucosal injury; IAP; NF-κB

中图分类号:

S816.7

陶燕子, 王秋菊, 袁玲, 崔一喆, 贾军峰. LPS致炎时间对小鼠小肠组织中IAP与NF-κB表达的影响[J]. 中国畜牧兽医, 2023, 50(11): 4424-4432.

TAO Yanzi, WANG Qiuju, YUAN Ling, CUI Yizhe, JIA Junfeng. Effect of LPS-induced Inflammatory Time on IAP and NF-κB Expression in Small Intestine of Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4424-4432.

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