大肠杆菌对奶牛子宫内膜上皮细胞炎性损伤的研究

doi:10.16431/j.cnki.1671-7236.2021.07.039

Abstract

Abstract: The purpose of this experiment was to study the in vitro inflammatory damage of bovine endometrial epithelial cells caused by Escherichia coli (E.coli),and explore the optimal concentration and duration time of E.coli to induce inflammatory reactions and its mechanism.Firstly,different concentrations of E.coli (5×10⁴,5×10⁵,1×10⁶,2.5×10⁶,5×10⁶ CFU/mL) were used to infect the cells for 3,6 and 9 h,then detected the effect of E.coli on cell viability by observing the cells morphology with inverted microscope morphology and measured the D_{450 nm} with CCK-8 method;Secondly,different concentrations of E.coli (5×10⁴,5×10⁵ CFU/mL) were used to infect the cells for 3,6 and 9 h,then detected the secretion levels of IL-1β,IL-6,IL-8 and TNF-α in the cell supernatant with ELISA method.Finally,different concentrations of E.coli (5×10⁴,5×10⁵ CFU/mL) were used to infect the cells for 6 and 9 h,then the phosphorylation level of IκBα and p65 protein were detected by Western blotting.The results showed that compared with control group,the cell viability of 1×10⁶,2.5×10⁶ and 5×10⁶ CFU/mL E.coli groups was extremely significantly reduced after 9 h of E.coli infection (P<0.01),while the cell viability of 5×10⁵ CFU/mL E.coli group was significantly decreased (P<0.05).The levels of IL-1β,IL-6,IL-8 and TNF-α of 5×10⁵ CFU/mL E.coli group were extremely significantly increassed after 9 h infection (P<0.01).Moreover,the levels of IκBα and p65 phosphorylation protein were significantly higher than those in both 5×10⁴ CFU/mL and control groups (P<0.01).The levels of IL-6,IκBα and p65 phosphorylation protein in 5×10⁵ CFU/mL E.coli group were extremely significantly increased after 6 h infection (P<0.01),while IκBα and p65 phosphorylation protein in the 5×10⁴ CFU/mL E.coli group were significantly increased after 6 h infection (P<0.05).In conclusion,bovine endometrial epithelial cells inflammatory response could be induced by E.coli and the optimal action time was 6 h when E.coli infection concentration was 5×10⁵ CFU/mL,and 9 h when E.coli infection concentration was 5×10⁴ CFU/mL.

Key words: bovine endometrial epithelial cells; Escherichia coli; inflammation; cytokines

CLC Number:

S823.9⁺1

LI Botong, WANG Shiyu, LIU Jiawei, ZHANG Weiqi, NI Yaodi, LIU Mingchao. Effect of Escherichia coli on Inflammatory Damage of Bovine Endometrial Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2627-2634.

References

[1] SHELDON I M,WILLIAMS E J,MILLER A N A,et al.Uterine diseases in cattle after parturition[J].Veterinary Journal,2008,176(1):115-121.
[2] WILLIAMS E J,FISCHER D P,PFEIFFER D U,et al.Clinical evaluation of postpartum vaginal mucus reflects uterine bacterial infection and the immune response in cattle[J].Theriogenology,2005,63(1):102-117.
[3] FÖLDI J,KULCSÁR M,PÉCSI A,et al.Bacterial complications of postpartum uterine involution in cattle[J].Animal Reproduction Science,2006,96(3-4):265-281.
[4] 刘昆,毛伟,李婷婷,等.大肠杆菌与金黄色葡萄球菌对奶牛子宫内膜组织细胞因子表达及损伤程度的影响[J].中国畜牧兽医,2019,46(6):1841-1848. LIU K,MAO W,LI T T,et al.Effect of Escherichia coli and staphylococcus aureus on expression of cytokine and damage degree of dairy cows endometrium tissue[J].China Animal Husbandry & Veterinary Medicine,2019,46(6):1841-1848.(in Chinese)
[5] BELL M J,ROBERTS D J.The impact of uterine infection on a dairy cow's performance[J].Theriogeno-logy,2007,68(7):1074-1079.
[6] 杜金梁,徐丽娜,马玉忠,等.家兔子宫内膜细胞炎症模型的建立[J].中国农学通报,2009,25(4):18-20. DU J L,XU L N,MA Y Z,et al.Inflammatory model of endometrial cells in rabbits[J].Chinese Agricultural Science Bulletin,2009,25(4):18-20.(in Chinese)
[7] 邝晓娇,张世栋,董书伟,等.丹参水提物对山羊子宫内膜上皮细胞炎症模型中基质金属蛋白酶-2表达的影响[J].动物医学进展,2015,36(3):54-59. KUANG X J,ZHANG S D,DONG S W,et al.Effect of extract of salvia miltiorrhiza on MMP-2 in endometrium epithelial cell inflammation model of goat[J].Progress in Veterinary Medicine,2015,36(3):54-59.(in Chinese)
[8] 杨倩,史万玉,赵驻军,等.小鼠子宫内膜细胞炎症模型的建立[J].河北农业大学学报,2011,34(6):87-90. YANG Q,SHI W Y,ZHAO Z J,et al.Inflammatory model of endometrial cells in mice[J].Journal of Agricultural University of Hebei,2011,34(6):87-90.(in Chinese)
[9] 于晓红,白云静,程鹏,等.苦参碱对奶牛子宫内膜上皮细胞抗炎作用及其机制[J].北京农学院学报,2015,30(3):35-39. YU X H,BAI Y J,CHENG P,et al.Anti-inflammatory effect of matrine on the bovine endometrial epithelial cell and its mechanism[J].Journal of Beijing University of Agriculture,2015,30(3):35-39.(in Chinese)
[10] 王仕宇,李博通,刘佳玮,等.原代奶牛子宫内膜上皮细胞体外培养方法的改进[J].中国畜牧兽医,2020,47(3):958-964. WANG S Y,LI B T,LIU J W,et al.Improvement of culture method of primary bovine endometrial epithelial cells in vitro[J].China Animal Husbandry & Veterinary Medicine,2020,47(3):958-964.(in Chinese)
[11] SHELDON I M,CRONIN J,GOETZE L,et al.Defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle[J].Biology of Reproduction,2009,81(6):1025-1032.
[12] 赵红霞.奶牛子宫内膜炎致病大肠杆菌的分离、鉴定及耐药性研究[D].呼和浩特:内蒙古农业大学,2008. ZHAO H X.Studies on identification and resistance of pathogenic E.coli isolated from bovine with endometritis[D].Hohhot:Inner Mongolia Agricultural University,2008.(in Chinese)
[13] 杭苏琴,王一昊,郑婕,等.产后奶牛子宫内大肠杆菌的分离和血清型鉴定[J].畜牧与兽医,2015,47(3):104-106. HANG S Q,WANG Y H,ZHENG J,et al.Isolation and identification of Escherichia coli from the postpartum dairy cows[J].Animal Husbandry & Veterinary Medicine,2015,47(3):104-106.(in Chinese)
[14] 赵立香,郭梦尧,李丰阳,等.大肠杆菌引发奶牛子宫内膜炎的致病机制研究[J].黑龙江畜牧兽医,2013,19:14-17. ZHAO L X,GUO M Y,LI F Y,et al.Research on the pathogenic mechanism of endometritis caused by Escherichia coli in dairy cattle[J].Heilongjiang Animal Science and Veterinary Medicine,2013,19:14-17.(in Chinese)
[15] 李德军,刘运枫,裴小英,等.子宫内膜炎急性相蛋白与IL-6变化的研究[J].畜牧兽医学报,2010,41(10):1333-1336. LI D J,LIU Y F,PEI X Y,et al.Research on change of acute phase protein and IL-6 in cows with endometritis[J].Chinese Journal of Animal and Veterinary Sciences,2010,41(10):1333-1336.(in Chinese)
[16] 刘明超.奶牛子宫内膜炎早期诊断及益生菌抗大肠杆菌诱导的子宫内膜上皮细胞炎性损伤作用机理[D].北京:中国农业大学,2017. LIU M C.Early diagnosis of endometritis of dairy cows and the mechanism of probiotics against inflammatory injury of bovine endometrial epithelial cells induced by Escherichia coli[D].Beijing:China Agricultural University,2017.(in Chinese)
[17] OECKINGHAUS A,HAYDEN M S,GHOSH S.Crosstalk in NF-κB signaling pathways[J].Nature Immunology,2011,12(8):695-708.
[18] FERNÁNDEZ M F,BORIS S,BARBÉS C.Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract[J].Journal of Applied Microbiology,2003,94(3):449-455.
[19] SMALE S T.Hierarchies of NF-κB target-gene regulation[J].Nature Immunology,2011,12(8):689-694.
[20] MEDZHITOV R.Origin and physiological roles of inflammation[J].Nature,2008,454(7203):428-435.
[21] YIN P,ZHANG Z,LI J,et al.Ferulic acid inhibits bovine endometrial epithelial cells against LPS-induced inflammation via suppressing NK-κB and MAPK pathway[J].Research in Veterinary Science,2019,126:164-169.
[22] 张桂林,崔晓妮,曹荣峰.脂多糖诱导奶牛子宫内膜上皮细胞核因子κB的表达[J].中国兽医科学,2010,40(10):1063-1066. ZHANG G L,CUI X N,CAO R F.Expression of NF-κB LPS-induced in bovine endometrial epithelial cell[J].Chinese Veterinary Science,2010,40(10):1063-1066.(in Chinese)
[23] WEI Z,WANG J,SHI M,et al.Saikosaponin a inhibits LPS-induced inflammatory response by inducing liver X receptor alpha activation in primary mouse macrophages[J].Oncotarget,2016,7(31):48995-49007.

Effect of Escherichia coli on Inflammatory Damage of Bovine Endometrial Epithelial Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	CHENG Qian, GAO Qingqing, WANG Yuhe, HUAN Changchao, GAO Song. Effects of Avian Pathogenic Escherichia coli Outer Membrane Protein OmpA on Autophagy of DF-1 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1971-1980.
[2]	XU Chenghui, ZHANG Xumei, YANG Tong, LI Jiahui, SUN Yawei, SHI Huijun, FU Qiang, YANG Li. Molecular Mechanism of Leonurine in the Treatment of Intestinal Inflammation Based on Network Pharmacology and Molecular Docking [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2103-2113.
[3]	SHAO Yongbin, ZHENG Yichuang, ZHANG Jingyi, WEI Linying, TIAN Yong, SUN Guojie, LIU Yuran, LUO Yan, GU Xinli. Study on the Anti-inflammatory Effect of Leonurine on LPS-induced Endometritis in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2149-2155.
[4]	WANG Leping, PAN Yan, TAN Yizhou, LI Lei, HAN Kaiou, CAO Yajie, LI Jun, ZHOU Yuqing, LIAO Yuying, MA Dongxin, GONG Yu, PENG Hao, WANG Xiaoye. Isolation,Identification and Genome Analysis of Escherichia coli Phage vB_EcoP_GN07 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1522-1531.
[5]	GUO Bingyu, LI Li, LI Chen, KONG Xianghui. Role and Molecular Mechanism of Mpeg1 in Pathogen Clearance [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1584-1592.
[6]	LI Cuihan, QU Zhina, GAO Yubin, LI Yan, WANG Juan, DUAN Xiaoxiao, WANG Lin, ZHANG Xiyue, ZHAO Ge, HUANG Xiumei, ZHAO Jianmei, ZHANG Qingqing, WANG Junwei, HUANG Baoxu, LIU Junhui. Epidemiology Investigation and Whole Genome Sequencing Analysis of Three Types of Poultry Originated Carbapenem-resistant Escherichia coli [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1653-1662.
[7]	HUANG Qiaochen, LIAO Binru, YU Chunyan, MO Yupeng, HUANG Weiqian, LI Lei, HAN Kaiou, WANG Leping, LI Xun, WANG Xiaoye. Isolation,Identification and Biological Characteristics Analysis of a Bacteriophage EP_H11 from Escherichia coli [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1684-1694.
[8]	ZHU Qiqi, ZHANG Jiyu, WANG Chengye, WU Lingyu, WANG Weiwei, ZHU Zhen. Analysis of Multidrug Resistance and Prevalence of Animal-derived Escherichia coli in Hebei [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1081-1092.
[9]	LIANG Tian, WANG Mingdi, SUN Peng, ZHU Aichen, ZHAO Qianhui, HUO Shuying. Network Pharmacological Analysis of the Anti-inflammatory Mechanism of Traditional Chinese Medicine Mixture Phellodendron amurense and Acorus calamus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1195-1206.
[10]	LUO Gan, ZOU Hong, REN Shaoke, WU Chunxia, ZOU Yao, LOU Yinying, ZHOU Yang, CHENG Fangjun, GUO Jianhua. Detection of O-antigen,Virulence Gene and Drug Sensitivity Test of Pathogenic Escherichia coli from Duck [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 713-721.
[11]	LIU Lin, WU Baoqing, GUO Xiaoyin, YU Wenhui, TIAN Lang. Study on Bacteriostatic Action of Aqueous Extract of Granati pericarpium Against Multi-drug Resistant Escherichia coli of Swine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 328-340.
[12]	LIU Haoyuan, LI Yanjie, QIN Hewei, JIN Xiaoqin, NIU Yuqing, HUA Xiaoqiong, ZHANG Shuqin, NIU Li. Study on the Mechanism of WU Shen Venation Smooth Decoction on Atherosclerosis AopE^-/- Mice Based on Network Pharmacology [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 377-389.
[13]	SHAO Mingzhu, ZHAO Yunqing, LI Songli, HOU Shaohua, ZHANG Yanying. Isolation and Identification of Escherichia coli from Dairy Cows in Hebei Province and Its Pathogenicity and Antibacterial Effect of Chinese Herbal Medicine in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3683-3691.
[14]	LU Wengeng, SI Linqing, TIAN Chunyu, YUAN Siqi, XIE Hexin, ZHOU Jiwei, JIN Jidong, HAN Yinghao. Establishment of Oxidative Stress Model of Bovine Endometrial Epithelial Cells and the Effects of Bovine Adipose-derived Mesenchymal Stem Cells on Cell Migration [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2506-2514.
[15]	DU Wenqi, XIA Liye, LI Guimei, SHAN Hu. Prokaryotic Expression and Purification of Seneca Virus A VP1 Protein [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2708-2715.