硫酸小檗碱对猪轮状病毒感染的IPEC-J2中TGF-β1、EGFR、EGF、IGF1 mRNA表达的影响

doi:10.11843/j.issn.0366-6964.2018.10.025

摘要/Abstract

摘要：

利用猪轮状病毒（PoRV）感染仔猪肠黏膜上皮细胞（IPEC-J2）建立损伤模型，研究硫酸小檗碱（BS）对PoRV感染的IPEC-J2中TGF-β₁、EGFR、EGF、IGF₁基因mRNA表达的影响。通过构建目的基因和内参基因的重组质粒，绘制荧光标准曲线，采用实时荧光定量PCR法测定BS对PoRV感染的IPEC-J2中TGF-β₁、EGFR、EGF、IGF₁ mRNA表达的影响。结果显示：PoRV能够显著引起IPEC-J2变性和坏死脱落，用BS药物组的IPEC-J2则生长基本良好；同时计算试验组相对于对照组各因子mRNA表达的差异倍数，显示BS有促进病毒损伤IPEC-J2中TGF-β₁和EGFR mRNA表达，抑制EGF和IGF₁ mRNA表达的作用。可见在肠黏膜细胞损伤修复过程中，BS能够显著促进TGF-β₁和EGFR基因表达，但是抑制EGF和IGF₁基因表达，表明BS对各细胞因子在修复过程中发挥的作用和时间存在明显差异。BS可能通过调节损伤的肠黏膜上皮细胞修复因子的基因表达，进而调控肠上皮细胞的增殖、转化，对受损肠黏膜起到保护和修复作用。

Abstract:

This study was conducted to investigate the effects of berberine sulfate (BS) on TGF-β₁, EGFR, EGF and IGF₁ expression in porcine intestinal epithelial cell (IPEC-J2) that were infected by rotavirus. We established a model in which IPEC-J2 were infected with rotavirus. We established target and reference gene recombinant plasmid and standard curve. TGF-β₁, EGFR, EGF and IGF₁ mRNA levels were quantified in BS and rotavirus treated IPEC-J2 by real-time polymerase chain reaction. Results were as follows:after the infection of control IPEC-J2 with rotavirus, the cells clustered into poorly-organized groups, shed or died. By contrast, the BS-treated IPEC-J2 that were infected with rotavirus formed highly-organized groups. The difference of TGF-β₁, EGFR, EGF and IGF₁ gene mRNA expression in each experimental group was compared with that of the control group, showing that BS can promote the expression of TGF-β₁ and EGFR. However, BS inhibited the expression of EGF and IGF₁ mRNA. Compared with EGF and IGF₁ gene expression decreased significantly, the expression of TGF-β₁ and EGFR gene were increased significantly, suggesting that the effect of repair factors may not be the same in the process of repairing cells injury. Our results show that BS could ameliorate IPEC-J2 injury by regulating repair factors expression to promote the proliferation and transformation of cells for repairing the intestinal mucosa.

中图分类号:

S853.7

徐环叶, 李妹玲, 陈立功, 王丽叶, 孟利佳, 崔欢, 张诚, 白晓, 王迎春, 董世山. 硫酸小檗碱对猪轮状病毒感染的IPEC-J2中TGF-β₁、EGFR、EGF、IGF₁ mRNA表达的影响[J]. 畜牧兽医学报, 2018, 49(10): 2283-2291.

XU Huan-ye, LI Mei-ling, CHEN Li-gong, WANG Li-ye, MENG Li-jia, CUI Huan, ZHANG Cheng, BAI Xiao, WANG Ying-chun, DONG Shi-shan. Berberine Sulfate Affects TGF-β₁, EGFR, EGF and IGF₁ mRNA Expression in Rotavirus-infected IPEC-J2[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(10): 2283-2291.

参考文献

[1] 邱福双. 断奶仔猪腹泻的原因及预防保健措施[J]. 畜牧与饲料科学, 2013, 34(2):112-114.
QIU F S. Causes and preventive health measures of diarrhea in weanedpiglets[J]. Animal Husbandry and Feed Science, 2013, 34(2):112-114. (in Chinese)
[2] 杨宾,黄群兰. 仔猪腹泻的病因及综合防治技术[J]. 山东畜牧兽医, 2015(5):23-25.
YANG B, HUANG Q L. Etiology and comprehensive control techniques of piglets'diarrhea[J]. Shandong Journal of Animal Science and Veterinary Medicine, 2015(5):23-25. (in Chinese)
[3] WANG N, TAN H Y, LI L, et al. Berberine and Coptidis Rhizoma as potential anticancer agents:Recent updates and future perspectives[J]. J Ethnopharmacol, 2015, 176:35-48.
[4] 张瑞芬,苏和. 黄连的药理研究进展[J]. 内蒙古中医药, 2010, 29(3):114-117.
ZHANG R F, SU H. Thepharmacological research of coptis[J]. Inner Mongol Journal of Traditional Chinese Medicine, 2010, 29(3):114-117. (in Chinese)
[5] 汪玉芳,柯善栋,陶秀良. 黄连素对人肝癌HepG2细胞增殖及血管内皮生长因子表达的影响[J]. 中国中西医结合消化杂志, 2013, 21(3):143-145.
WANG Y F, KE S D, TAO X L.Effects of berberine on the expression of vascular endothelial growth factor in HepG2 cells[J]. Chinese Journal of Integrated Traditional and Western Medicine on Digestion, 2013, 21(3):143-145. (in Chinese)
[6] 李妹玲,乔宁,陈立功,等. 仔猪肠黏膜上皮细胞体外培养及其对小檗碱适应性的研究[J]. 河北农业大学学报, 2016, 39(1):99-102.
LI M L, QIAO N, CHEN L G, et al.The culture of piglet intestinal epithelial cells in vitro and its adaptability to berberine[J]. Journal of Agricultural University of Hebei, 2016, 39(1):99-102. (in Chinese)
[7] LI Y J, TIAN D A. Correlation between apoptosis and TGF-β1 expression in the mucosal epithelium of rat small intestine in a cold stress state[J]. Exp Ther Med, 2013, 5(5):1456-1460.
[8] 郑明洁,王秀利,常亚青,等. 表皮生长因子家族及其在海胆中的研究进展[J]. 生物技术通报, 2008(2):31-34, 38.
ZHENG M J, WANG X L, CHANG Y Q, et al. Epidermal growth factor family andtheir study advances in sea urchin[J]. Biotechnology Bulletin, 2008(2):31-34, 38. (in Chinese)
[9] 吴国柱,郭跃先,杨士杰. 胰岛素样生长因子-1(IGF-1)的临床应用进展[J]. 河北医药, 2015, 37(9):1398-1400.
WU G Z, GUO Y X, YANG S J. Clinical application of insulin-like growth factor-1(IGF-1)[J]. Hebei Medical Journal, 2015, 37(9):1398-1400. (in Chinese)
[10] OLIVEIRA S R, VIEIRA H L A, DUARTE C B. Effect of carbon monoxide on gene expression in cerebrocortical astrocytes:Validation of reference genes for quantitative real-time PCR[J]. Nitric Oxide, 2015, 49:80-89.
[11] 姜文灿,岳素文,江洪,等. TaqMan探针法实时荧光定量PCR的应用和研究进展[J]. 临床检验杂志:电子版, 2015, 4(1):797-805.
JIANG W C, YUE S W, JIANG H, et al. Application and research progress of TaqMan probe real time PCR[J]. Clinical Laboratory Journal:Electronic Edition, 2015, 4(1):797-805. (in Chinese)
[12] 李融. pcDNA3. 0-SV40T重组质粒的构建及其在鹅小肠上皮细胞中的表达[D]. 扬州:扬州大学, 2009.
LI R.Construction of recombinant plasmid pcDNA3.0-SV40T and expression in goose intestinal epithelial cells[D]. Yangzhou:Yangzhou University, 2009. (in Chinese)
[13] 李丽,赵成萍,李宏,等. 质粒制备绝对定量PCR标准曲线方法的建立[J]. 农业生物技术学报, 2011, 19(6):1157-1162.
LI L, ZHAO C P, LI H, et al.Establishment of the plasmid standard curve generation method for absolute quantification PCR[J]. Journal of Agricultural Biotechnology, 2011, 19(6):1157-1162. (in Chinese)
[14] GUERRERO C A, ACOSTA O. Inflammatory and oxidative stress in rotavirus infection[J]. World J Virol, 2016, 5(2):38-62.
[15] TISSERA M S, COWLEY D, BOGDANOVIC-SAKRAN N, et al. Options for improving effectiveness of rotavirus vaccines in developing countries[J]. Hum Vaccin Immunother, 2016, 13(4):921-927.
[16] 徐倩倩. 白头翁素对小鼠PRV、E. coli混合感染性腹泻防治机制的研究[D]. 保定:河北农业大学, 2010.
XU Q Q. Studies on prevention and cure mechanism of anemonin on diarrhea in mice induced with PRV and E. coli mixed infection[D]. Baoding:Hebei Agricultural University, 2010. (in Chinese)
[17] ZHU C, WU Y P, JIANG Z Y, et al. Dietary soy isoflavone attenuated growth performance and intestinal barrier functions in weaned piglets challenged with lipopolysaccharide[J]. Int Immunopharmacol, 2015, 28(1):288-294.
[18] 张志杨. 溃疡性结肠炎患者结肠粘膜中转化生长因子-β亚型的临床表达[J]. 临床医药文献杂志, 2016, 3(36):7166-7167.
ZHANG Z Y. Clinical expression of transforming growth factor-β isoforms in colonic mucosa of patients with ulcerative colitis[J]. Journal of Clinical Medical, 2016, 3(36):7166-7167. (in Chinese)
[19] 林源,周勇杰,廖书胜,等. 健胃愈疡颗粒对消化性溃疡患者胃黏膜TGF-β1 mRNA表达的影响[J]. 实用预防医学, 2007, 14(2):306-308.
LIN Y, ZHOU Y J, LIAO S S, et al. Effect of Jianwei Yuyang granule on expression of TGF-β1 mRNA in gastric mucosa in patients with gastric ulcer[J]. Practical Preventive Medicine, 2007, 14(2):306-308. (in Chinese)
[20] 刘密,常小荣,严洁,等. 艾灸对胃黏膜损伤大鼠胃黏膜表皮生长因子、转化生长因子-α及其受体的影响[J]. 针刺研究, 2011, 36(6):403-408.
LIU M, CHANG X R, YAN J, et al.Effects of moxibustion on gastric mucosal EGF and TGF-α contents and epidermal growth factor receptor expression in rats with gastric mucosal lesion[J]. Acupuncture Research, 2011, 36(6):403-408. (in Chinese)
[21] 田云霄,张小辉. 表皮生长因子受体及其与胃癌的关系研究[J]. 实用心脑肺血管病杂志, 2012, 20(5):920-922.
TIAN Y X, ZHANG X H. Epidermal growth factor receptor and its relationship with gastriccarcinoma[J]. Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease, 2012, 20(5):920-922. (in Chinese)
[22] 刘颖,徐倩倩,薛晓阳,等. 白头翁素对腹泻小鼠肠黏膜修复因子EGFR和TGFβ1的影响[J]. 畜牧兽医学报, 2013, 44(3):475-480.
LIU Y, XU Q Q, XUE X Y, et al. Effect of anemonin on intestinal mucosa repair factors EGFR and TGFβ1 of diarrheal mice[J]. Acta Veterinaria et Zootechnica Sinica, 2013, 44(3):475-480. (in Chinese)
[23] 张志勇,孙公凯,杨世忠. 表皮生长因子在消化性溃疡中的研究进展[J]. 海南医学院学报, 2010, 16(3):388-390.
ZHANG Z Y, SUN G K, YANG S Z. Research progress of epidermal growth factor in pepticulcer[J]. Journal of Hainan Medical University, 2010, 16(3):388-390. (in Chinese)
[24] 顾玲. 胆胃宁颗粒对湿热中阻型胃溃疡的治疗作用及EGF、PGE₂含量的影响[D]. 南京:南京中医药大学, 2014.
GU L. Effect of Dan-Wei-Ning granules on gastric ulcer of damp-heat in energizer and the content of EGF, PGE₂[D]. Nanjing:Nanjing University of Chinese Medicine, 2014. (in Chinese)
[25] 宋丹,方秀斌. 表皮生长因子的研究进展[J]. 解剖科学进展, 2002, 8(4):339-342.
SONG D, FANG X B.Advances of epidermal growth factor[J]. Progress of Anatomical Sciences, 2002, 8(4):339-342. (in Chinese)
[26] 程亚颖,宋光耀,周利霞,等. 不同出生胎龄的2~7岁小于胎龄儿生长激素释放多肽、胰岛素样生长因子-1、胰岛素水平及其与生长发育的关系[J]. 卫生研究, 2012, 41(1):18-22.
CHENG Y Y, SONG G Y, ZHOU L X, et al. Relationship of plasma ghrelin, IGF-1 and insulin with the growth and development of 2-7 year-old children with small for gestational age at birth[J]. Journal of Hygiene Research, 2012, 41(1):18-22. (in Chinese)
[27] CLEMMONS D R. Role of IGF binding proteins in regulating metabolism[J]. Trends Endocrinol Metab, 2016, 27(6):375-391.
[28] 卢劲晔,王锦峰,卢炜,等. 胰岛素样生长因子1研究进展[J]. 畜牧与兽医, 2014, 46(5):116-118.
LU J Y, WANG J F, LU W, et al. Research progress of insulin-like growth factor-1[J]. Animal Husbandry & Veterinary Medicine, 2014, 46(5):116-118. (in Chinese)
[29] ARMAKOLAS N, ARMAKOLAS A, ANTONOPOULOS A, et al. The role of the IGF-1 Ec in myoskeletal system and osteosarcoma pathophysiology[J]. Crit Rev Oncol Hematol, 2016, 108:137-145.
[30] BALLESTER M, CORDÍN R, FOLCH J M. DAG expression:High-throughput gene expression analysis of real-time PCR data using standard curves for relative quantification[J]. PLoS One, 2013, 8(11):e80385.
[31] GOSSELIN-THÉBERGE M, TABOADA E, GUY R A. Evaluation of real-time PCR assays and standard curve optimisation for enhanced accuracy in quantification of Campylobacter environmental water isolates[J]. J Microbiol Methods, 2016, 129:70-77.
[32] KUMAR M, NANDI S. Development of a SYBR Green based real-time PCR assay for detection and quantitation of canine parvovirus in faecal samples[J]. J Virol Methods, 2010, 169(1):198-201.

硫酸小檗碱对猪轮状病毒感染的IPEC-J2中TGF-β₁、EGFR、EGF、IGF₁ mRNA表达的影响

Berberine Sulfate Affects TGF-β₁, EGFR, EGF and IGF₁ mRNA Expression in Rotavirus-infected IPEC-J2

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