苦苣菜对肺炎小鼠炎症因子及HMGB1/TLR4/NF-κB信号通路的影响

doi:10.16431/j.cnki.1671-7236.2023.12.044

摘要/Abstract

摘要： 【目的】研究苦苣菜水提物对脂多糖(LPS)诱导的小鼠肺炎模型中炎症相关因子及炎症信号通路的影响,探究苦苣菜水提物对小鼠肺炎的抗炎作用。【方法】制备苦苣菜水提物,将昆明小鼠随机分为空白组、模型组、阳性对照组及苦苣菜水提物低、中、高浓度组,空白组小鼠不做任何处理,模型组小鼠灌胃0.3 mL生理盐水,阳性对照组小鼠灌胃0.3 mL 5 mg/kg地塞米松,苦苣菜水提物低、中、高浓度组小鼠依次灌胃0.3 mL 100、200和400 mg/mL苦苣菜水提物,持续灌胃给药1周后,空白组小鼠腹腔注射0.3 mL生理盐水,其余组小鼠腹腔注射0.3 mL 30 mg/kg LPS诱导建立小鼠肺炎模型。采用ELISA方法检测各组小鼠血清中白细胞介素-6(IL-6)、IL-10、转化生长因子-β(TGF-β)的含量;利用免疫组织化学(IHC)和Western blotting检测各组小鼠肺中炎症信号通路(HMGB1/TLR4/NF-κB)蛋白的表达水平。【结果】与空白组相比,模型组小鼠血清中IL-6含量显著上调(P＜0.05),IL-10、TGF-β含量有所上调,肺脏中HMGB1、TLR4、NF-κB的分布及蛋白表达量极显著上调(P＜0.01);与模型组相比,苦苣菜水提物处理组小鼠血清中促炎因子IL-6含量有所下调,抗炎因子IL-10含量有所上调,抗炎因子TGF-β含量极显著上调(P＜0.01);在肺脏中HMGB1、TLR4、NF-κB大量分布在肺泡周围的上皮细胞上,与模型组相比,苦苣菜水提物处理组小鼠肺脏中HMGB1、TLR4、NF-κB的分布极显著下调(P＜0.01),且表现出剂量依赖效应,HMGB1、TLR4蛋白表达量极显著下调(P＜0.01),NF-κB蛋白表达量显著或极显著下调(P＜0.05;P＜0.01)。【结论】苦苣菜水提物能抑制LPS诱导的肺炎小鼠血清中促炎因子IL-6的分泌及肺脏中信号通路HMGB1、TLR4、NF-κB蛋白的表达,促进小鼠血清中抗炎因子IL-10、TGF-β的分泌,表明苦苣菜水提物具有抗肺炎作用。

关键词: 苦苣菜; 肺炎; 炎症因子; HMGB1/TLR4/NF-κB

Abstract: 【Objective】 The aim of this study was to investigate the effects of Sonchus oleraceus L.aqueous extract on inflammation-related factors and inflammatory signaling pathways in mouse pneumonia model induced by lipopolysaccharide (LPS),to explore the anti-inflammatory effects of Sonchus oleraceus L.aqueous extract on mouse pneumonia.【Method】 Preparation of aqueous extract of Sonchus oleraceus L.,the test mice were randomly divided into blank group,model group,positive control group,and Sonchus oleraceus L.aqueous extract low,medium and high concentrations groups,mice in blank group were not treated,mice in model group were gavaged with 0.3 mL of saline,mice in positive control group were gavaged with 0.3 mL of 5 mg/kg dexamethasone,and mice in Sonchus oleraceus L.aqueous extract low,medium and high concentrations groups were gavaged with 0.3 mL of 100,200 and 400 mg/mL Sonchus oleraceus L.aqueous extract,respectively.After being administered by gavage for 1 week,mice in blank group were injected intraperitoneally with 0.3 mL of saline,and mice in the rest of groups were injected intraperitoneally with 0.3 mL of 30 mg/kg LPS to induce the establishment of a mouse pneumonia model. The levels of interleukin-6 (IL-6),IL-10 and transforming growth factor-β (TGF-β) in serum of each group were measured by ELISA,and the expression levels of inflammatory signaling pathway (HMGB1/TLR4/NF-κB) proteins in lungs of mice in each group were measured by IHC and Western blotting.【Result】 Compared with blank group,the serum levels of IL-6 in mice were significantly up-regulated (P＜0.05),the levels of IL-10 and TGF-β in mice were up-regulated,and the distribution and expression of HMGB1,TLR4 and NF-κB proteins in lung of mice were highly significantly up-regulated in model group (P＜0.01).Compared with model group,the serum levels of pro-inflammatory factor IL-6 in mice was down-regulated,anti-inflammatory factor IL-10 was up-regulated,and anti-inflammatory factor TGF-β was highly significantly up-regulated in Sonchus oleraceus L.aqueous extract-treated groups (P＜0.01).HMGB1,TLR4 and NF-κB was abundantly distributed on the epithelial cells surrounding the alveoli in lungs.Compared with model group,the distribution of HMGB1,TLR4 and NF-κB were highly significantly down-regulated in lung of mice treated with aqueous extract of Sonchus oleraceus L.(P＜0.01),and showed a dose-dependent effect,the protein expression of HMGB1 and TLR4 were exremely significantly or down-regulation (P＜0.01),and the protein expression of NF-κB was significantly or extremely significantly down-regulated (P＜0.05 or P＜0.01).【Conclusion】 The aqueous extract of Sonchus oleraceus L.could inhibit the secretion of pro-inflammatory factor IL-6 and the expression of signaling pathway HMGB1,TLR4 and NF-κB in lung of mice with pneumonia induced by intraperitoneal injection of LPS,and promote the secretion of anti-inflammatory factor IL-10 and TGF-β in serum of mice,indicating that the aqueous extract of Sonchus oleraceus L.had anti-pneumonia effect.

Key words: Sonchus oleraceus L.; pneumonia; inflammatory factors; HMGB1/TLR4/NF-κB

中图分类号:

S852.21

李孟阳, 晁冰, 周涛, 张建萍. 苦苣菜对肺炎小鼠炎症因子及HMGB1/TLR4/NF-κB信号通路的影响[J]. 中国畜牧兽医, 2023, 50(12): 5232-5242.

LI Mengyang, CHAO Bing, ZHOU Tao, ZHANG Jianping. Effect of Sonchus oleraceus L.on Inflammatory Factors and HMGB1/TLR4/NF-κB Signaling Pathway in Mice with Pneumonia[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5232-5242.

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