RESEARCH PAPER

Baicalin Alleviates Lipopolysaccharide Induced Jejunal Inflammation and Oxidative Stress via Inhibiting Nuclear Factor-κB and Activating Nuclear Factor E2-Related Factor 2 Signaling Pathways in Mice

  • BAO Minglong ,
  • SUN Xinyi ,
  • LIANG Mei ,
  • JU Xianghong ,
  • YONG Yanhong ,
  • MA Xingbin ,
  • YU Zhichao ,
  • GUO Yiying ,
  • LIU Xiaoxi
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  • Department of Veterinary Medicine, College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang 524000, China

Received date: 2021-06-20

  Online published: 2022-02-15

Abstract

This study mainly explored the alleviation effect and molecular mechanisms of baicalin on lipopolysaccharide (LPS) induced jejunal inflammation and oxidative stress in mice. Thirty-six mice were randomly divided into control group, negative control group, model group, low-dose baicalin group, medium-dose baicalin group and high-dose baicalin group with six mice in each group. Mice in the control group did not make any treatment, mice in the negative control group and model group were separately treated by continuous 7 days gavage with 0.2 mL phosphate buffer saline (PBS), and mice in baicalin groups were separately treated by continuous 7 days gavage with 0.2 mL different doses of baicalin (low-dose group, 100 mg/kg BW; medium-dose group, 200 mg/kg BW; high-dose group, 400 mg/kg BW) solution, followed by intraperitoneal injection of 3.5 mg/kg BW LPS to construct the jejunal inflammation model of mice in the model group and the baicalin groups. The mice were sacrificed 24 h later, and the jejunum was harvested for morphological observation, and the expression level changes and the contents of inflammatory factors in the jejunum, and the expression level changes of key proteins of nuclear factor-κB (NF-κB) and nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathways in the jejunum were detected. The results showed that compared with the control group, the jejunal tissue of mice in the model group had damage to the villus structure, and the villus height to crypt depth ratio was significant decreased (P<0.05), the Toll-like receptor 4 (TLR4) mRNA expression level in the jejunum was significant increased (P<0.05), the inflammatory factors interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) mRNA expression levels and contents were significantly increased (P<0.05), the P65 protein expression level, and the ratios of phosphorylated inhibitor of nuclear factor-kappa Bα (P-IκBα)/inhibitor of nuclear factor-kappa Bα (IκBα) and phosphorylated P65 (P-P65)/P65 were significantly increased (P<0.05), and the superoxide dismutase(SOD) activity and HO-1 protein expression level were significantly reduced (P<0.05). Relative to the model group, the histological structure of the jejunum in mice was obviously improved in the baicalin groups, with a significant higher ratio of villus height to crypt depth, and the mRNA expression levels of TLR4 and inflammatory factors (P<0.05), and the protein expression levels of P-65 and P-IκBα were significantly decreased (P<0.05); the SOD activity in the jejunum in high-dose baicalin group was significantly increased (P<0.05); the mRNA expression levels of HO-1 and quinone oxidoreductase-1 (NQO-1)in the jejunum in the high-dose baicalin group and medium-dose baicalin group were significantly increased (P<0.05), and the protein expression levels of nuclear Nrf2 and HO-1 were significantly elevated (P<0.05). In conclusion, in a jejunal inflammation model of mice introduced by LPS, baicalin at an optimum dose can exert anti-inflammatory function by inhibiting NF-κB pathway and play antioxidant function by activating Nrf2 signaling pathway. The recommended dose is 200 mg/kg BW.

Cite this article

BAO Minglong , SUN Xinyi , LIANG Mei , JU Xianghong , YONG Yanhong , MA Xingbin , YU Zhichao , GUO Yiying , LIU Xiaoxi . Baicalin Alleviates Lipopolysaccharide Induced Jejunal Inflammation and Oxidative Stress via Inhibiting Nuclear Factor-κB and Activating Nuclear Factor E2-Related Factor 2 Signaling Pathways in Mice[J]. Chinese Journal of Animal Nutrition, 2022 , 34(2) : 1217 -1229 . DOI: 10.3969/j.issn.1006-267x.2022.02.053

References

[1] CHAN G, FARZAN A, SOLTES G, et al.The epidemiology of Clostridium perfringens type A on Ontario swine farms, with special reference to cpb2-positive isolates[J].BMC Veterinary Research, 2012, 8:156.
[2] 范长友.刺五加多糖对脂多糖刺激断奶仔猪肠粘膜损伤的影响研究[D].硕士学位论文.沈阳:沈阳农业大学, 2020. FAN C Y.Effects of Acanthopanax senticosus polysaccharides on intestinal mucosal injury in immune stress weaned piglets[D].Master's Thesis.Shenyang:Shenyang Agricultural University, 2020.(in Chinese)
[3] 易宗容, 冯堂超.饲粮添加白藜芦醇对断奶仔猪生长性能和肠道健康的影响[J].中国饲料, 2020(10):32-36. YI Z R, FENG T C.Effects of dietary resveratrol supplementation on growth performance and intestinal health of weaned piglets[J].China Feed, 2020(10):32-36.(in Chinese)
[4] ZENG Y D, WANG Z R, ZOU T D, et al.Bacteriophage as an alternative to antibiotics promotes growth performance by regulating intestinal inflammation, intestinal barrier function and gut microbiota in weaned piglets[J].Frontiers in Veterinary Science, 2021, 8:623899.
[5] FANDY T E, JIEMJIT A, THAKAR M, et al.Decitabine induces delayed reactive oxygen species (ROS) accumulation in leukemia cells and induces the expression of ROS generating enzymes[J].Clinical Cancer Research, 2014, 20(5):1249-1258.  
[6] 朱凌羽, 张子琪, 兰海楠, 等.虾青素对脂多糖通过TLR4/MyD88/NF-κB信号通路诱导的IPEC-J2细胞炎症的影响[J].华南农业大学学报, 2018, 39(5):53-58. ZHU L Y, ZHANG Z Q, LAN H N, et al.Effects of astaxanthin on IPEC-J2 cell inflammation induced by lipopolysaccharide via TLR4/MyD88/NF-κB signaling pathway[J].Journal of South China Agricultural University, 2018, 39(5):53-58.(in Chinese)
[7] ZHOU J Y, LIN H L, QIN Y C, et al.L-carnosine protects against deoxynivalenol-induced oxidative stress in intestinal stem cells by regulating the Keap1/Nrf2 signaling pathway[J/OL].Molecular Nutrition & Food Research, 2021:e2100406.[2021-07-03].https://pubmed.ncbi.nlm.nih.gov/34216418/.DOI:10.1002/mnfr.202100406.
[8] MENG X L, HU L, LI W Q.Baicalin ameliorates lipopolysaccharide-induced acute lung injury in mice by suppressing oxidative stress and inflammation via the activation of the Nrf2-mediated HO-1 signaling pathway[J].Naunyn-Schmiedeberg's Archives of Pharmacology, 2019, 392(11):1421-1433.  
[9] WANG J, ISHFAQ M, LI J C.Baicalin ameliorates Mycoplasma gallisepticum-induced inflammatory injury in the chicken lung through regulating the intestinal microbiota and phenylalanine metabolism[J].Food & Function, 2021, 12(9):4092-4104.  
[10] QIAN Y Z, CHEN Y, WANG L Y, et al.Effects of baicalin on inflammatory reaction, oxidative stress and PKDl and NF-κB protein expressions in rats with severe acute pancreatitis1[J].Acta Cirurgica Brasileira, 2018, 33(7):556-564.  
[11] WANG X X, CHANG X H, ZHAN H B, et al.Curcumin and baicalin ameliorate ethanol-induced liver oxidative damage via the Nrf2/HO-1 pathway[J/OL].Journal of Food Biochemistry, 2020:e13425.[2020-08-08].https://pubmed.ncbi.nlm.nih.gov/32770697/.DOI:10.1111/jfbc.13425.
[12] 刘晓曦, 董杰, 李敏霞, 等.黄芩水提液对肠炎模型小鼠空肠损伤修复的作用[J].畜牧兽医学报, 2020, 51(2):392-398. LIU X X, DONG J, LI M X, et al.Effect of the Scutellaria baicalensis water extraction on the regulation of jejunum damage and repair in the model of enteritis mice[J].Acta Veterinaria et Zootechnica Sinica, 2020, 51(2):392-398.(in Chinese)
[13] YI Q Y, LIU J X, ZHANG Y F, et al.Anethole attenuates enterotoxigenic Escherichia coli-induced intestinal barrier disruption and intestinal inflammation via modification of TLR signaling and intestinal microbiota[J].Frontiers in Microbiology, 2021, 12:647242.
[14] 杨志远, 苏洁, 俞静静, 等.铁皮石斛超微粉对LPS致肠黏膜损伤小鼠的保护作用研究[J].中国中药杂志, 2021, 46(7):1667-1673. YANG Z Y, SU J, YU J J, et al.Protective effects of Dendrobium officinale superfine powder against LPS-induced intestinal mucosal injury in mice[J].China Journal of Chinese Materia Medica, 2021, 46(7):1667-1673.(in Chinese)
[15] 王梦竹.京尼平苷对LPS诱导小鼠肠道损伤时NF-κB/IAP的影响[D].硕士学位论文.大庆:黑龙江八一农垦大学, 2020. WANG M Z.Effect of geniposide on NF-κB/IAP in LPS-induced intestinal damage in mice[D].Master's Thesis.Daqing:Heilongjiang Bayi Agricultural University, 2020.(in Chinese)
[16] HUANG Y Q, SUN M Y, YANG X F, et al.Baicalin relieves inflammation stimulated by lipopolysaccharide via upregulating TUG1 in liver cells[J].Journal of Physiology and Biochemistry, 2019, 75(4):463-473.  
[17] LIU X Y, WANG S L, ZHAO G A.Baicalin relieves lipopolysaccharide-evoked inflammatory injury through regulation of miR-21 in H9c2 cells[J].Phytotherapy Research, 2020, 34(5):1134-1141.  
[18] 汪旭, 张鑫, 马晓娟, 等.黄芩苷在脂多糖(LPS)诱导的大鼠多器官急性损伤中的保护作用[J].河南科技大学学报(医学版), 2019, 37(2):104-107, 123. WANG X, ZHANG X, MA X J, et al.Protective effect of baicalin on LPS-induced acute injury in multiple rat organs[J].Journal of Henan University of Science & Technology (Medical Science), 2019, 37(2):104-107, 123.(in Chinese)
[19] 张莹.诃子提取物对LPS致小鼠肠黏膜损伤的保护作用及机制研究[D].硕士学位论文.哈尔滨:东北农业大学, 2019. ZHANG Y.Protective effect and its mechanism of Chebulia extract on intestinal mucosal injury induced by LPS in mice[D].Master's Thesis.Harbin:Northeast Agricultural University, 2019.(in Chinese)
[20] 王忠清, 林春发, 钟文杰, 等.术芩提取液对脂多糖损伤的IPEC-J2细胞增殖及炎症因子转录的影响[J].畜牧兽医学报, 2019, 50(7):1500-1508. WANG Z Q, LIN C F, ZHONG W J, et al.Effect of Zhuqin extractive fluid on the proliferation and transcription of inflammatory factors in LPS-injured IPEC-J2 cells[J].Acta Veterinaria et Zootechnica Sinica, 2019, 50(7):1500-1508.(in Chinese)
[21] 刘晓曦, 马云飞, 李焕荣, 等.加味葛根芩连汤对湿热泄泻仔猪肠道炎症和损伤修复的作用[J].畜牧兽医学报, 2021, 52(1):238-247. LIU X X, MA Y F, LI H R, et al.Effect of Jiawei Gegen Qinlian decoction on the regulation of intestine damage and repair in the piglets of damp-heat diarrhea[J].Acta Veterinaria et Zootechnica Sinica, 2021, 52(1):238-247.(in Chinese)
[22] WULLAERT A.Role of NF-kappaB activation in intestinal immune homeostasis[J].International Journal of Medical Microbiology, 2010, 300(1):49-56.  
[23] BURKEY T E, SKJOLAAS K A, DRITZ S S, et al.Expression of Toll-like receptors, interleukin 8, macrophage migration inhibitory factor, and osteopontin in tissues from pigs challenged with Salmonella enterica serovar Typhimurium or serovar Choleraesuis[J].Veterinary Immunology and Immunopathology, 2007, 115(3/4):309-319.
[24] MAO X H, SU Z Y, MOOKHTIAR A K.Long non-coding RNA:a versatile regulator of the nuclear factor-κB signalling circuit[J].Immunology, 2017, 150(4):379-388.  
[25] 王宇, 陈霞, 吴立斌, 等.艾灸对腹泻型肠易激综合征模型大鼠海马与结肠组织中IKKβ/IKBα/NF-κB通路的影响[J].安徽中医药大学学报, 2020, 39(3):32-36. WANG Y, CHEN X, WU L B, et al.Effect of moxibustion on the IKKβ/IKBα/NF-κB pathway in hippocampal and colonic tissue in a rat model of diarrhea-predominant irritable bowel syndrome[J].Journal of Anhui University of Chinese Medicine, 2020, 39(3):32-36.(in Chinese)
[26] LIU M J, SONG S X, LI H R, et al.The protective effect of caffeic acid against inflammation injury of primary bovine mammary epithelial cells induced by lipopolysaccharide[J].Journal of Dairy Science, 2014, 97(5):2856-2865.  
[27] LIU X X, LIU F H, MA Y F, et al.Effect of puerarin, baicalin and berberine hydrochloride on the regulation of IPEC-J2 cells infected with enterotoxigenic Escherichia coli[J].Evidence-Based Complementary and Alternative Medicine, 2019, 2019:7438593.
[28] 王欢.虾青素对赭曲霉毒素A引起小鼠空肠损伤的影响[D].硕士学位论文.沈阳:沈阳农业大学, 2020. WANG H.Effects of astaxanthin on ochratoxin A-induced jejunum damage in mice[D].Master's Thesis.Shenyang:Shenyang Agricultural University, 2020.(in Chinese)
[29] SIES H.Oxidative stress:oxidants and antioxidants[J].Experimental Physiology, 1997, 82(2):291-295.  
[30] YIN J, WU M M, XIAO H, et al.Development of an antioxidant system after early weaning in piglets[J].Journal of Animal Science, 2014, 92(2):612-619.  
[31] YUAN X, ZHOU Y, WANG W, et al.Activation of TLR4 signaling promotes gastric cancer progression by inducing mitochondrial ROS production[J].Cell Death & Disease, 2013, 4(9):e794.
[32] GOU Z Y, JIANG S Q, ZHENG C T, et al.Equol inhibits LPS-induced oxidative stress and enhances the immune response in chicken HD11 macrophages[J].Cellular Physiology and Biochemistry, 2015, 36(2):611-621.  
[33] FAN H, LE J W, SUN M, et al.Pretreatment with S-nitrosoglutathione attenuates septic acute kidney injury in rats by inhibiting inflammation, oxidation, and apoptosis[J].BioMed Research International, 2021, 2021:6678165.
[34] 王思博, 崔红霞, 杨季, 等.超氧化物歧化酶模拟物对氧化应激IPEC-J2细胞抗氧化性能的影响[J].中国畜牧兽医, 2021, 48(5):1566-1573. WANG S B, CUI H X, YANG J, et al.Effect of superoxide dismutase mimetic on antioxidant capacity of oxidatively damaged IPEC-J2 cells[J].China Animal Husbandry & Veterinary Medicine, 2021, 48(5):1566-1573.(in Chinese)
[35] LI L R, DONG H, SONG E Q, et al.Nrf2/ARE pathway activation, HO-1 and NQO1 induction by polychlorinated biphenyl quinone is associated with reactive oxygen species and PI3K/AKT signaling[J].Chemico-Biological Interactions, 2014, 209:56-67.
[36] HEO H S, HAN G E, WON J, et al.Pueraria montana var. lobata root extract inhibits photoaging on skin through Nrf2 pathway[J].Journal of Microbiology and Biotechnology, 2019, 29(4):518-526.  
[37] ISHFAQ M, CHEN C L, BAO J X, et al.Baicalin ameliorates oxidative stress and apoptosis by restoring mitochondrial dynamics in the spleen of chickens via the opposite modulation of NF-κB and Nrf2/HO-1 signaling pathway during Mycoplasma gallisepticum infection[J].Poultry Science, 2019, 98(12):6296-6310.  
[38] 朱慧渊, 万海同, 王江, 等.中医药防治缺血性脑卒中剂量-效应关系的探讨[J].中医药学报, 2021, 49(6):9-11. ZHU H Y, WAN H T, WANG J, et al.Dose-effect relationship in preventing and treating cerebral ischemic stroke with traditional Chinese medicine[J].Acta Chinese Medicine and Pharmacology, 2021, 49(6):9-11.(in Chinese)
[39] 刘显军, 陈静, 王丽, 等.断奶仔猪试验中建立LPS应激模型的作用体现[J].黑龙江畜牧兽医, 2017(17):141-143. LIU X J, CHEN J, WANG L, et al.Effects of LPS stress model in weaning piglet test[J].Heilongjiang Animal Science and Veterinary Medicine, 2017(17):141-143.(in Chinese)
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