豆腐果苷对脂多糖诱导的猪小肠上皮细胞损伤分析

doi:10.6048/j.issn.1001-4330.2022.10.029

新疆农业科学 ›› 2022, Vol. 59 ›› Issue (10): 2595-2600.DOI: 10.6048/j.issn.1001-4330.2022.10.029

• 植物保护·微生物·农业生态环境·畜牧兽医 • 上一篇

豆腐果苷对脂多糖诱导的猪小肠上皮细胞损伤分析

石乐天¹(), 陈勇臻², 郭腾达¹, 刘金铄¹, 张光辉¹(), 陈鹏举³^,⁴()

1.河南农业大学动物医学院,郑州 450002
2.焦作工学院,河南焦作,454000
3.河南省现代中兽医研究院,郑州 450002
4.山东鑫德慧生物科技有限公司,山东郓城 274700

收稿日期:2022-01-10 出版日期:2022-10-20 发布日期:2022-12-21
通信作者: 张光辉,陈鹏举
作者简介:石乐天(1998-),男,硕士研究生,研究方向为兽用新制剂研发与兽医公共卫生,(E-mail)1989798043@qq.com
基金资助:
国家重点研发计划“畜禽重大疫病防控与高效安全养殖综合技术研发”重点专项“新型动物药剂创制与产业化”(2017YFD0501400)

The Protective Effect of Helicid on Lipopolysaccharide Induced Intestinal Epithelial Cell Injury in Pigs

SHI Letian¹(), CHEN Yongzhen², GUO Tengda¹, LIU Jinshuo¹, ZHANG Guanghui¹(), CHEN Pengju³^,⁴()

1. College of Veterinary Medicine, Henan Agricultural University, ZhengZhou, 450002
2. Jiaozuo Institute of Technology, JiaoZuo,454000
3. Henan Institute of Modern Chinese Veterinary Medicine,ZhengZhou,450002
4. Shandong Xindehui Biotechnology Co.,Ltd, Yuncheng, Shandong Province 274700, China

Received:2022-01-10 Online:2022-10-20 Published:2022-12-21
Correspondence author: ZHANG Guanghui, CHEN Pengju
Supported by:
National Key R & D Program Project “Comprehensive Research and Development of Prevention and Control of Livestock and Poultry Epidemic Prevention and Control and Efficient and Safe Breeding—Creation and Industrialization of New Animal Agents”(2017YFD0501400)

摘要/Abstract

摘要：

【目的】研究脂多糖(LPS)诱导猪空肠上皮细胞(IPEC-J2)来建立氧化及炎症模型,分析豆腐果苷(HEL)抗炎及抗氧化的作用机理。【方法】试验分为5组:对照组,LPS组,HEL(25、50和100 μM)+LPS组。【结果】HEL会显著降低炎性因子TNF-α、IL-1β和IL-6在mRNA水平的表达;LPS会显著提高氧化相关蛋白(Nrf2和HO-1)的表达和抗氧化酶(SOD和CAT)的活性,显著减少氧化产物(MDA)的含量,HEL表现出相反的结果,显著降低了Nrf2和HO-1的表达量和SOD和CAT的活性,显著增加了MDA的含量。【结论】HEL会缓解LPS所引起的IPEC-J2细胞的氧化及炎性损伤,一定程度上保护细胞免受伤害。

关键词: HEL; LPS; 炎症; 氧化损伤; IPEC-J2细胞

Abstract:

【Objective】 The aim of this study is to establish an oxidative and inflammatory model of porcine jejunal epithelial cells (IPEC-J2) induced by lipopolysaccharide (LPS), and explore the anti-inflammatory and anti-oxidation mechanism of helicid (HEL).【Methods】 The experiment was divided into 5 groups: control group, LPS group, HEL (25, 50 and 100 μM) + LPS group.【Results】 The results showed that HEL could significantly reduce the expression of TNF-α, IL-1β and IL-6 at mRNA level, and we further verified the anti-inflammatory mechanism of HEL at protein level by ELISA.In terms of oxidative damage, LPS could significantly improve the expression of oxidation related proteins (Nrf2 and HO-1) and the activity of antioxidant enzymes (SOD and CAT), and significantly reduce the content of oxidation products (MDA).Importantly, HEL showed the opposite results, reducing the expression of Nrf2, HO-1 and the activity of SOD and CAT significantly and also significantly increasing the content of MDA.【Conclusion】 In conclusion, all the results show that HEL can alleviate the oxidative and inflammatory damage of IPEC-J2 cells caused by LPS, and protect cells from injury to a certain extent.

Key words: HEL; LPS; inflammation; oxidative damage; IPEC-J2 cells

中图分类号:

S828

石乐天, 陈勇臻, 郭腾达, 刘金铄, 张光辉, 陈鹏举. 豆腐果苷对脂多糖诱导的猪小肠上皮细胞损伤分析[J]. 新疆农业科学, 2022, 59(10): 2595-2600.

SHI Letian, CHEN Yongzhen, GUO Tengda, LIU Jinshuo, ZHANG Guanghui, CHEN Pengju. The Protective Effect of Helicid on Lipopolysaccharide Induced Intestinal Epithelial Cell Injury in Pigs[J]. Xinjiang Agricultural Sciences, 2022, 59(10): 2595-2600.

图/表 5

表1 引物序列

Table 1 Primer sequence

引物名称 Primer name	引物序列(5’-3’) Primer sequence (5 '- 3')
TNF-α	F:CGACTCAGTGCCGAGATCAA
TNF-α	R:CCTGCCCAGATTCAGCAAAG
IL-1β	F:TGATTGTGGCAAAGGAGGA
IL-1β	R:TTGGGTCATCATCACAGACG
IL-6	F:CCAGGAACCCAGCTATGAAC
IL-6	R:CTGCACAGCCTCGACATT
GAPDH	F:ACATCATCCCTGCTTCTACTGG
GAPDH	R:CTCGGACGCCTGCTTCAC

图1 HEL和LPS细胞毒性变化

Fig.1 Effects of Hel and LPS on the cytotoxicity of IPEC-J2 cells

图2 不同添加剂量HEL下TNF-α、IL-1β和IL-6表达水平变化

Fig.2 Effect of HEL on the expression of inflammatory factors

图3 不同添加剂量HEL下SOD,CAT和MDA变化

Fig.3 Effect of HEL on SOD, CAT and MDA

图4 不同添加剂量HEL下TNF-α、IL-1β、IL-6、Nrf2和HO-1蛋白变化

Fig.4 Effects of HEL on TNF-α, IL-1 β, IL-6, Nrf2 and HO-1 proteins

参考文献 21

[1]	Tang J, Jiang Y, Tang Y, et al. Effects of propofol on damage of rat intestinal epithelial cells induced by heat stress and lipopolysaccharides[J]. Brazilian Journal of Medical & Biological Research, 2013, 46(6):507-512.
[2]	Renzhammer R, Loncaric I, Roch F F, et al. Prevalence of Virulence Genes and Antimicrobial Resistances in E.coli Associated with Neonatal Diarrhea, Postweaning Diarrhea, and Edema Disease in Pigs from Austria[J]. Antibiotics, 2020, 09(4).
[3]	Diao X, Man L, Cheng X, et al. Identification of metabolites of helicid in vivo using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry[J]. Biomedical Chromatography, 2018, 32:4263.
[4]	Liu G Y, Shuang C, Zhang Y M, et al. Study on chemical constituents in seeds of Helicia nilagirica (II)[J]. China Journal of Chinese Materia Medica, 2005, 30(11):830-832.
[5]	Wei Y, Cao R, Wen H, et al. Synthesis and biological evaluation of helicid analogues as mushroom tyrosinase inhibitors[J]. Bioorganic & Medicinal Chemistry Letters, 2008, 18(24):6490-6493. DOI URL
[6]	Tong J C K.Lü, Yang B, et al. Effects of helicid on hippocampal neurogenesis in chronically stressed mice[J]. Chinese Pharmaceutical Journal, 2012, 47(7):512-516.
[7]	Li X Y, Qi W W, Zhang Y X, et al. Helicid Ameliorates Learning and Cognitive Ability and Activities cAMP/PKA/CREB Signaling in Chronic Unpredictable Mild Stress Rats[J]. Biological and Pharmaceutical Bulletin, 2019, 42(7):1146-1154. DOI URL
[8]	Rong Y, Han X, Dai X C. Studies on antidepressant activity of helicid[J]. Pharmacology and Clinics of Chinese Materia Medica, 2007, 23:22-23.
[9]	Tong J, Zhou Z, Qi W, et al. Antidepressant effect of helicid in chronic unpredictable mild stress model in rats[J]. International Immunopharmacology, 2019, 67:13-21. DOI PMID
[10]	袁媛, 孙梅. 黄芪多糖对LPS损伤小肠上皮细胞的保护作用[J]. 世界华人消化杂志, 2008(1):15-19.
	YUAN Yuan, SUN Mei. Mechanism of impact of Astragalus mongholicus polysaccharides on lipopolysaccharide-iinduced damage in intestinal epithelial cells[J]. World Chinese Journal of Digestology, 2008(1):15-19.
[11]	钟佳, 卞艺斐, 刘萍, 等. 橙皮苷抑制LPS诱导的肠上皮细胞炎性损伤及其机制研究[J]. 中国兽医杂志, 2022, 058(001):66-74.
	ZHONG Jia, BIAN Yifei, LIU Ping, et al. Effect and Mechanism of Hesperidin on Lipopolysaccharide-Induced Inflammatory Injury of Intestinal Epithelial Cells[J]. Chinese Journal of Veterinary Medicine, 2022, 058(001):66-74.
[12]	Piwowarski J P, Kiss A K, Granica S, et al. Urolithins, gut microbiota-derived metabolites of ellagitannins, inhibit LPS-induced inflammation in RAW264.7 murine macrophages[J]. Molecular Nutrition & Food Research, 2015, 59(11):2168-2177.
[13]	Spitzer A J, Tian Q, Choudhary R K, et al. Bacterial Endotoxin Induces Oxidative Stress and Reduces Milk Protein Expression and Hypoxia in the Mouse Mammary Gland[J]. Oxidative Medicine and Cellular Longevity, 2020, 2020(6):1-16.
[14]	Mz A, Mhb D, Hs C, et al. Vitamin D 3 attenuates lipopolysaccharide-induced cognitive impairment in rats by inhibiting inflammation and oxidative stress[J]. Life Sciences, 2020.
[15]	Loffredo L, Ivanov V, Ciobanu N, et al. Is there an association between atherosclerotic burden, oxidative stress and gut-derived lipopolysaccharides?[J]. Antioxid Redox Signal, 2020.
[16]	林谦, 邱磊, 云龙, 等. 核因子E2相关因子2调控机体抗氧化途径特性及其与畜禽的健康和肉品质的关系[J]. 动物营养学报, 2014, 26(6):1421-1429.
	LIN Qian, QIU Lei, YUN Long, et al. Nuclear Factor Erythroid-2-Related Factor 2 Mediated Antioxidant Pathway Character and Its Relation on Health and Meat Quality of Livestock and Poultry[J]. Chinese Journal of Animal Nutrition, 2014, 26(6):1421-1429.
[17]	Ryter S W, Otterbein L E, Morse D, et al. Heme oxygenase/carbon monoxide signaling path-ways: Regulation and functional significance[J]. Molecular and Cellular Biochemistry, 2002, 234-235.
[18]	Smale S T, Natoli G. Transcriptional control of inflammatory responses[J]. Cold Spring Harbor Perspectives in Biology, 2014, 6(11):a016261. DOI URL
[19]	Medzhitov R, Horng T. Transcriptional control of the inflammatory response[J]. Nature Reviews Immunology, 2009, 9(10):692-703. DOI PMID
[20]	Zhang Y, Luan D, Li X, et al. Helicid Reverses Lipopolysaccharide-Induced Inflammation and Promotes GDNF Levels in C6 Glioma Cells through Modulation of Prepronociceptin[J]. Chemistry & Biodiversity, 2020, 17(7).
[21]	Chen S, Zhang C L, Zhou X F, et al. Anti-inflammatory and antioxidative properties of helicid protect against CCl 4 induced acute liver injury in mice[J]. Biotechnic and Histochemistry, 2020:1-7.

豆腐果苷对脂多糖诱导的猪小肠上皮细胞损伤分析

The Protective Effect of Helicid on Lipopolysaccharide Induced Intestinal Epithelial Cell Injury in Pigs

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