RESEARCHPAPER

Effects of Sophora alopecuroides L. on c-Jun N-Terminal Kinase/p38 Mitogen-Activated Protein Kinase Signaling Pathway and Tight Junction Protein Expression in Rumen Epithelium of Fattening Lambs Fed High Concentrate Diet

  • LI Shufang ,
  • MA Tian ,
  • YANG Jinli ,
  • AN Yawen ,
  • ZHANG Yu ,
  • YANG Xiaodong ,
  • WANG Hairong
Expand
  • Inner Mongolia Key Laboratory of Animal Nutrition and Feed Science, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2021-11-29

  Online published: 2022-06-14

Abstract

The purpose of this experiment was to explore the effects of Sophora alopecuroides L. on c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (p38MAPK) signaling pathway and tight junction protein expression in rumen epithelium of fattening lambs fed high concentrate diet. Thirty-two Dumont lambs with the body weight of (25.73±2.17) kg were randomly divided into 4 groups with 8 lambs in each group. Lambs in control group was fed with a high concentrate basal diet (concentrate:forage=7:3), and others in test groups were fed the basal diet supplemented with 0.1%, 0.3% and 0.5% Sophora alopecuroides L., which were named T1, T2 and T3 groups, respectively. The feeding experiment including 15-day of pretrial period and 60-day of formal trial period. All lambs were slaughtered at the end of formal trial period. The rumen epithelial tissue was sampled. The morphology of rumen epithelium was detected by hematoxylin-eosin staining method, the mRNA relative expression levels of inflammatory factor and tight junction protein genes in rumen epithelium were detected by real-time PCR (RT-PCR), the inflammatory factor contents in rumen epithelium were detected by enzyme-linked immunosorbent assay (ELISA) method, and the relative expression levels of JNK/p38MAPK signaling pathway and tight junction proteins were detected by Western blotting method. The results showed as follows:1) compared with the control group, the abnormal morphology of the rumen epithelial tissue such as keratinization, abnormal shedding, and nipple rupture in the test groups was improved. The rumen papillae length in test groups was significantly higher than that in the control group (P<0.05), and the muscle thickness in T1 and T2 groups was extremely significantly higher than that in the control group (P<0.01). 2) Compared with the control group, the mRNA relative expression levels of interferon-γ (IFN-γ), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) in the T1 group were significantly or extremely significantly down-regulated (P<0.05 or P<0.01), and the IFN-γ content was significantly decreased (P<0.05); the mRNA relative expression levels of IFN-γ, interleukin-1β (IL-1β) and TNF-α in the T2 group were significantly down-regulated (P<0.05); the mRNA relative expression level of IL-10 in the T1 and T2 groups was extremely significantly up-regulated (P<0.01), and the interleukin-10 (IL-10) content in the T1 group was extremely significantly increased (P<0.01). 3) Compared with the control group, the mRNA and protein relative expression levels of claudin-1 and occluding in T1 group were significantly or extremely significantly up-regulated (P<0.05 or P<0.01), and the mRNA relative expression level of zonula occluden-1 (ZO-1) was significantly up-regulated (P<0.05). 4) Compared with the control group, the phosphorylation level of JNK protein in T1 group was significantly reduced (P<0.05), and the phosphorylation level of p38MAPK protein in T1 and T2 groups was significantly reduced (P<0.05). In conclusion, on the one hand, the appropriate amount of Sophora alopecuroides L. can inhibit the phosphorylation of JNK and p38MAPK proteins, regulate the production and release of inflammatory factors, and alleviate the rumen damage caused by high concentrate diet; on the other hand, the appropriate amount of Sophora alopecuroides L. may directly regulate the expression of the tight junction proteins, reduce the permeability of rumen mucosa and enhance the rumen epithelial barrier function.

Cite this article

LI Shufang , MA Tian , YANG Jinli , AN Yawen , ZHANG Yu , YANG Xiaodong , WANG Hairong . Effects of Sophora alopecuroides L. on c-Jun N-Terminal Kinase/p38 Mitogen-Activated Protein Kinase Signaling Pathway and Tight Junction Protein Expression in Rumen Epithelium of Fattening Lambs Fed High Concentrate Diet[J]. Chinese Journal of Animal Nutrition, 2022 , 34(6) : 3940 -3952 . DOI: 10.3969/j.issn.1006-267x.2022.06.053

References

[1] PENNER G B, OBA M, GÄBEL G, et al.A single mild episode of subacute ruminal acidosis does not affect ruminal barrier function in the short term[J].Journal of Dairy Science, 2010, 93(10):4838-4845.  
[2] 孙燕勇.亚急性瘤胃酸中毒对奶山羊瘤胃上皮通透性的影响及其机制研究[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2017. SUN Y Y.Mechanism research on rumen epithelial permeability by subacute ruminal acidosis in dairy goats[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2017.(in Chinese)
[3] 杨淑青.亚急性瘤胃酸中毒对奶山羊瘤胃上皮屏障功能影响机制的研究[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2014. YANG S Q.Study on effect of subacute ruminal acidosis on rumen epithelial barrier function in dairy goats[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2014.(in Chinese)
[4] 唐志文, 孙福昱, 杨亮, 等.不同饲粮条件下奶牛胃肠道中内毒素浓度与炎症反应相关关系研究进展[J].家畜生态学报, 2018, 39(5):6-10. TANG Z W, SUN F Y, YANG L, et al.Concentrations of endotoxin in gastrointestinal tract of dairy cows under different dietary conditions and the relationship with inflammation[J].Acta Ecologae Animalis Domastici, 2018, 39(5):6-10.(in Chinese)
[5] OPAL S M.The host response to endotoxin, antilipopolysaccharide strategies, and the management of severe sepsis[J].International Journal of Medical Microbiology, 2007, 297(5):365-377.  
[6] LU Y C, YEH W C, OHASHI P S.LPS/TLR4 signal transduction pathway[J].Cytokine, 2008, 42(2):145-151.  
[7] LI S, KHAFIPOUR E, KRAUSE D O, et al.Effects of subacute ruminal acidosis challenges on fermentation and endotoxins in the rumen and hindgut of dairy cows[J].Journal of Dairy Science, 2012, 95(1):294-303.  
[8] 游菁菁, 李月英, 沙碧莹, 等.中药苦豆子生物碱的研究进展[J].江西中医药大学学报, 2015, 27(2):109-113, 116. YOU J J, LI Y Y, SHA B Y, et al.Advances in studies on the alkaloid of Sophora alopecuroides L.[J].Journal of Jiangxi University of Traditional Chinese Medicine, 2015, 27(2):109-113, 116.(in Chinese)
[9] ZHOU H F, LI J Y, SUN F, et al.A review on recent advances in aloperine research:pharmacological activities and underlying biological mechanisms[J].Frontiers in Pharmacology, 2020, 11:538137.
[10] 韩燕, 周娅, 刘泉.苦豆子抗内毒素效应的实验研究[J].中药材, 2006, 29(10):1066-1069. HAN Y, ZHOU Y, LIU Q.Antiendotoxic effects of Sophora alopecuroides L.[J].Jorunal of Chinese Medicinal Materials, 2006, 29(10):1066-1069.(in Chinese)
[11] 关亮俊.苦豆子化学成分分析及槐定碱、13, 14-去氢槐定碱制备工艺研究[D].硕士学位论文.北京:北京中医药大学, 2019. GUAN L J.Chemical composition analysis of sophora alopecia and preparation of sophoridine, 13, 14-dehydrosophoridine[D].Master's Thesis.Beijing:Beijing University of Chinese Medicine, 2019.(in Chinese)
[12] 谢明欣.苦豆子对蒙古羔羊体外发酵参数、微生物种群及血液中生理生化指标的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2018. XIE M X.Effect of Sophora alopecuroides L. on rumen fermentation parameters, ruminal microbial population and blood physiological-biochemical indexes of Mongolian lamb[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2018.(in Chinese)
[13] 杨晓东.高精料日粮中添加苦豆子对蒙古羔羊生长性能、血清生化及免疫指标的影响.[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2020. YANG X D.Effect of adding Sophora alopecuroides L. in high-concentration diet on growth performance, serum biochemical and immune indexes of Mongolian lambs[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2020.(in Chinese)
[14] STEELE M A, GREENWOOD S L, CROOM J, et al.An increase in dietary non-structural carbohydrates alters the structure and metabolism of the rumen epithelium in lambs[J].Canadian Journal of Animal Science, 2012, 92(2):123-130.  
[15] 邬宇航.亚急性瘤胃酸中毒对瘤胃、瓣胃上皮细胞增殖与凋亡的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2013. WU Y H.Effects of subacute ruminal acidosis on proliferation and apoptosis of rumen and omasum epithelial cell[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2013.(in Chinese)
[16] 高智雄.日粮不同精粗比对羔羊生长性能和免疫机能的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2020. GAO Z X.Effects of different dietary concentrate to forage ration on growth performance and immune function of lambs[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2020.(in Chinese)
[17] SUN X D, YUAN X, CHEN L, et al.Histamine induces bovine rumen epithelial cell inflammatory response via NF-κB pathway[J].Cellular Physiology and Biochemistry, 2017, 42(3):1109-1119.  
[18] 许超.烟酸调控高精料诱导瘤胃上皮细胞凋亡机制的研究[D].硕士学位论文.南昌:江西农业大学, 2017. XU C.Regulation mechanism of nicotinic acid on apoptosis of rumen epithelial cell induced by high concentrate diets in vivo[D].Master's Thesis.Nanchang:Jiangxi Agricultural University, 2017.(in Chinese)
[19] JIA Y Q, YUAN Z W, ZHANG X S, et al.Total alkaloids of Sophora alopecuroides L. ameliorated murine colitis by regulating bile acid metabolism and gut microbiota[J].Journal of Ethnopharmacology, 2020, 255:112775.
[20] ZHANG Z Q, LIU Q F, ZANG H, et al.Oxymatrine protects against L-arginine-induced acute pancreatitis and intestine injury involving Th1/Th17 cytokines and MAPK/NF-κB signalling[J].Pharmaceutical Biology, 2019, 57(1):595-603.  
[21] 宁夏回族自治区卫生厅.宁夏中药材标准[M].银川:宁夏人民出版社, 1993:88. Health Department of Ningxia Hui Autonomous Region.Ningxia medicinal material standard[M].Yinchuan:Ningxia People's Publishing House, 1993:88.(in Chinese)
[22] 曹晓东.苦豆子总碱的毒性及HPLC法测定绵羊血浆四种苦豆子碱的研究[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2014. CAO X D.TASA toxicology and by HPLC method to detect four kinds of alkaloids in plasma of sheep[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2014.(in Chinese)
[23] 姚雯, 孙培环, 徐晓敏, 等.苦豆子种子生物碱类化学成分及细胞毒活性[J].中国实验方剂学杂志, 2014, 20(22):95-99. YAO W, SUN P H, XU X M, et al.Alkaloids constituents of sophora alopecuroides seed and their cytotoxic activity[J].Chinese Journal of Experimental Traditional Medical Formulae, 2014, 20(22):95-99.(in Chinese)
[24] TURNER J R.Molecular basis of epithelial barrier regulation:from basic mechanisms to clinical application[J].The American Journal of Pathology, 2006, 169(6):1901-1909.  
[25] 李碧波.绒山羊胃肠道微生物区系及其对日粮响应的研究[D].博士学位论文.杨凌:西北农林科技大学, 2020. LI B B.Study on gastro-intestinal microbial flora and its response to diet in cashmere goat[D].Ph.D.Thesis.Yangling:Northwest A & F University, 2020.(in Chinese)
[26] 邓波波, 冯宝宝, 詹康, 等.短链脂肪酸对奶牛瘤胃上皮细胞促炎因子、趋化因子和紧密连接蛋白表达量的影响[J].甘肃农业大学学报, 2020, 55(4):22-28. DENG B B, FENG B B, ZHAN K, et al.Effects of short-chain fatty acids on the expression of pro-inflammatory factors, chemokines and tight junction proteins in dairy cow rumen epithelial cells[J].Journal of Gansu Agricultural University, 2020, 55(4):22-28.(in Chinese)
[27] 姜勇, 罗深秋.细胞信号转导的分子基础与功能调控[M].北京:科学出版社, 2005. JIANG Y, LUO S Q.Molecular basis and functional regulation of cellular signal transduction[M].Beijing:Science Press, 2005.(in Chinese)
[28] JOHNSON G L, LAPADAT R.Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases[J].Science, 2002, 298(5600):1911-1912.  
[29] ONO K, HAN J.The p38 signal transduction pathway:activation and function[J].Cellular Signalling, 2000, 12(1):1-13.  
[30] KIM B, BRETON S.The MAPK/ERK-signaling pathway regulates the expression and distribution of tight junction proteins in the mouse proximal epididymis[J].Biology of Reproduction, 2016, 94(1):22.
[31] ZHANG K, MENG M J, GAO L P, et al.Rumen-derived lipopolysaccharide induced ruminal epithelium barrier damage in goats fed a high-concentrate diet[J].Microbial Pathogenesis, 2019, 131:81-86.
[32] GUO J F, CHANG G J, ZHANG K, et al.Rumen-derived lipopolysaccharide provoked inflammatory injury in the liver of dairy cows fed a high-concentrate diet[J].Oncotarget, 2017, 8(29):46769-46780.  
[33] GITTER A H, BENDFELDT K, SCHMITZ H, et al.Epithelial barrier defects in HT-29/B6 colonic cell monolayers induced by tumor necrosis factor-α[J].Annals of the New York Academy of Sciences, 2000, 915(1):193-203.
[34] WATSON C J, HOARE C J, GARROD D R, et al.Interferon-gamma selectively increases epithelial permeability to large molecules by activating different populations of paracellular pores[J].Journal of Cell Science, 2005, 118(Pt.22):5221-5230.
[35] 许霓珊, 曹惠慧, 卢子滨, 等.苦豆碱激活胆碱能通路发挥抗结肠炎作用[J].中国药理学与毒理学杂志, 2021, 35(10):752. XU N S, CAO H H, LU Z B, et al.Oxymatrine activates cholinergic pathway to play an anti-colitis role[J].Chinese Journal of Pharmacology and Toxicology, 2021, 35(10):752.(in Chinese)
Outlines

/