术苦芩总多糖对湿热泄泻仔猪小肠杯状细胞数量以及MUC-2和ITF-3 mRNA转录的影响
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摘要
为探讨术苦芩总多糖(ZKQPs)对湿热泄泻仔猪小肠修复作用的影响,将60头断奶仔猪分为空白对照组(n=10)造模组(n=50),造模组采用高温高湿环境配合高脂配合饲料等建立湿热泄泻模型,进一步将造模成功后的50头仔猪随机分为模型组、阳性药物组(白头翁散)、ZKQPs高、中、低剂量组(n=10),分别拌料给药,连用7 d。采用过碘酸雪夫染色(PAS)结合显微镜观察小肠组织杯状细胞(GC),利用RT-PCR和ELISA分别检测湿热泄泻仔猪小肠黏液中黏蛋白-2(MUC-2)和肠三叶因子(ITF-3)的转录和含量变化。试验结果:1)模型组仔猪十二指肠、空肠、回肠中GC数量均减少,与空白组比较,差异显著(P<0.05);与模型组比较,ZKQPs高剂量组、中剂量组和阳性药物组仔猪十二指肠、空肠、回肠中GC数量均增加,ZKQPs高剂量组、阳性药物组仔猪十二指肠、空肠、回肠和ZKQPs中剂量组仔猪十二指肠、回肠GC的数量差异显著(P<0.05)。2)与空白组比较,模型组仔猪十二指肠、空肠、回肠中MUC-2 mRNA的转录量和含量均降低,差异显著(P<0.05或P<0.01)。与模型组比较,ZKQPs各剂量组和阳性药物组仔猪十二指肠、空肠、回肠中MUC-2 mRNA的转录量和含量均升高,ZKQPs高剂量组和阳性药物组仔猪十二指肠、空肠、回肠MUC-2 mRNA的转录量和含量差异显著(P<0.05或P<0.01)。3)与空白组比较,模型组仔猪十二指肠、空肠、回肠中ITF-3 mRNA的转录量和含量均降低,差异极显著(P<0.01)。与模型组比较,ZKQPs高剂量组、中剂量组和阳性药物组仔猪十二指肠、空肠、回肠中ITF-3 mRNA的转录量和含量均升高,ZKQPs高剂量组、中剂量组和阳性药物组仔猪空肠、回肠中ITF-3 mRNA的转录量和含量差异显著(P<0.05或P<0.01)。ZKQPs可提高湿热泄泻仔猪小肠GC的数量以及MUC-2和ITF-3 mRNA的转录,促进湿热泄泻仔猪小肠内GC分泌物MUC-2和ITF-3分泌。
To investigate the effect of Polysaccharides from ZhuKuQin(ZKQPs) on the recovery function in damp-heat diarrhea Piglet intestine, 60 piglets were divided into blank control group(n=10) and damp-heat diarrhea model group(high temperature, humidity environment combined with high fat compound feed), and 50 model piglets were randomly divided into model group, positive drug group(Pulsatilla powder), ZKQPs high, middle and low dose group, with 10 piglets in each group, drugs were mixed with feed, for 7 days. Small intestinal goblet cells(GC) were stained with PAS and observed by Microscope, RT-PCR and ELISA were used to detect the transcriptional quantity and content of mucoprotein-2(MUC-2) and intestinal trilobular factor(ITF-3) in intestinal mucus of damp-heat diarrhea piglets. The results were as follows: 1)The number of GC in duodenum, jejunum and ileum of piglets in model group decreased significantly when compared with that in blank group(P<0.05); The number of GC in duodenum, jejunum and ileum of piglets in ZKQPs high, middle dose group and positive drug group increased. The number of GC in duodenum, jejunum and ileum of piglets in the ZKQPs high dose group and the positive drug group were significantly different when compared with the model group(P<0.05), and the number of GC in duodenum and ileum of piglets in ZKQPs middle dose group was significantly different(P<0.05). 2)The transcriptional quantity and content of MUC-2 in duodenum, jejunum and ileum of piglets were decreased in model group, and the difference was significant when compared with that of blank group(P<0.05).The transcriptional quantity and content of MUC-2 in duodenum, jejunum and ileum of piglets in each ZKQPs dose groups and positive drug groups increased. The transcriptional quantity and content of MUC-2 in duodenum, jejunum and ileum of piglets in ZKQPs high dose group and positive drug group were significantly different from those in model group(P<0.01). 3)The transcriptional quantity and content of ITF-3 in duodenum, jejunum and ileum of piglets were decreased in model group, and the difference was significant when compared with that of blank group(P<0.05). The transcriptional quantity and content of ITF-3 in duodenum, jejunum and ileum of piglets in ZKQPs high, middle dose group and positive drug group increased.The transcriptional quantity and content of ITF-3 in jejunum and ileum of piglets in ZKQPs high, middle dose group and positive drug group were significantly different from those in model group(P<0.01). ZKQPs can increase the number of GC and the transcription of MUC-2 and ITF-3 mRNA in small intestine of damp-heat diarrhea Piglets, and promote the secretion of MUC-2 and ITF-3 in small intestine of damp-heat diarrhea Piglets.
To investigate the effect of Polysaccharides from ZhuKuQin(ZKQPs) on the recovery function in damp-heat diarrhea Piglet intestine, 60 piglets were divided into blank control group(n=10) and damp-heat diarrhea model group(high temperature, humidity environment combined with high fat compound feed), and 50 model piglets were randomly divided into model group, positive drug group(Pulsatilla powder), ZKQPs high, middle and low dose group, with 10 piglets in each group, drugs were mixed with feed, for 7 days. Small intestinal goblet cells(GC) were stained with PAS and observed by Microscope, RT-PCR and ELISA were used to detect the transcriptional quantity and content of mucoprotein-2(MUC-2) and intestinal trilobular factor(ITF-3) in intestinal mucus of damp-heat diarrhea piglets. The results were as follows: 1)The number of GC in duodenum, jejunum and ileum of piglets in model group decreased significantly when compared with that in blank group(P<0.05); The number of GC in duodenum, jejunum and ileum of piglets in ZKQPs high, middle dose group and positive drug group increased. The number of GC in duodenum, jejunum and ileum of piglets in the ZKQPs high dose group and the positive drug group were significantly different when compared with the model group(P<0.05), and the number of GC in duodenum and ileum of piglets in ZKQPs middle dose group was significantly different(P<0.05). 2)The transcriptional quantity and content of MUC-2 in duodenum, jejunum and ileum of piglets were decreased in model group, and the difference was significant when compared with that of blank group(P<0.05).The transcriptional quantity and content of MUC-2 in duodenum, jejunum and ileum of piglets in each ZKQPs dose groups and positive drug groups increased. The transcriptional quantity and content of MUC-2 in duodenum, jejunum and ileum of piglets in ZKQPs high dose group and positive drug group were significantly different from those in model group(P<0.01). 3)The transcriptional quantity and content of ITF-3 in duodenum, jejunum and ileum of piglets were decreased in model group, and the difference was significant when compared with that of blank group(P<0.05). The transcriptional quantity and content of ITF-3 in duodenum, jejunum and ileum of piglets in ZKQPs high, middle dose group and positive drug group increased.The transcriptional quantity and content of ITF-3 in jejunum and ileum of piglets in ZKQPs high, middle dose group and positive drug group were significantly different from those in model group(P<0.01). ZKQPs can increase the number of GC and the transcription of MUC-2 and ITF-3 mRNA in small intestine of damp-heat diarrhea Piglets, and promote the secretion of MUC-2 and ITF-3 in small intestine of damp-heat diarrhea Piglets.
引文
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[3] 黄蓉,欧希龙.肠道黏膜屏障功能损伤机制及其防治的研究进展[J].现代医学,2015,43(5):659-662.HUANG R,OU X L.Research progress in the mechanism of intestinal mucosal barrier function and its prevention and treatment[J].Modern Medical Journal,2015,43(5):659-662.(in Chinese)
[4] KESIMER M,KIRKHAM S,PICKLES R J,et al.Tracheobronchial air-liquid interface cell culture:a model for innate mucosal defense of the upper airways[J].Am J Physiol Lung Cell Mol Physiol,2009,296(1):L92-L100.
[5] 李娟,吴丽华,石岩,等.MUC1,MUC2和MUC3在胃增生性息肉中的转录模式[J].世界华人消化杂志,2011,19(35):3591-3596.LI J,WU L H,SHI Y,et al.Expression patterns of MUC1,MUC2 and MUC3 in gastric hyperplastic polyps[J].World Chinese Journal of Digestology,2011,19(35):3591-3596.(in Chinese)
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[7] 董文逍,曹海龙,王邦茂.杯状细胞在肠道疾病发病中作用的研究进展[J].国际消化病杂志,2015,35(4):244-246,269.DONG W X,CAO H L,WANG B M.Progress in the role of goblet cells in the pathogenesis of intestinal diseases[J].International Journal of Digestive Diseases,2015,35(4):244-246,269.(in Chinese)
[8] SUEMORI S,LYNCH-DEVANEY K,PODOLSKY D K.Identification and characterization of rat intestinal trefoil factor:tissue- and cell-specific member of the trefoil protein family[J].Proc Natl Acad Sci U S A,1991,88(24):11017-11021.
[9] 曾宇君,赖建彬,朱兆荣,等.术苦芩颗粒对湿热泻痢仔猪血清D-木糖、淀粉酶及脂肪酶的影响[J].中国兽医杂志,2015,51(1):57-59.ZENG Y J,LAI J B,ZHU Z R,et al.The effect of Zhukuqin ganules on serum D-xylose,amylase and lipase in piglets of damp-heat and diarrhea syndrome[J].Chinese Journal of Veterinary Medicine,2015,51(1):57-59.(in Chinese)
[10] 庞敏,朱兆荣,乔芊芊,等.术芩总多糖对免疫低下小鼠脾淋巴细胞增殖与自噬的影响[J].畜牧兽医学报,2018,49(12):2762-2770.PANG M,ZHU Z R,QIAO Q Q,et al.Effect of polysaccharides from Zhuqin formula on the proliferation and autophagy of spleen lymphocyte in Immunocompromised Mice[J].Acta Veterinaria et Zootechnica Sinica,2018,49(12):2762-2770.(in Chinese)
[11] 赵祥升,董娜,冯锦东,等.益智多糖含量测定[J].中国现代应用药学,2013,30(10):1070-1074.ZHAO X S,DONG N,FENG J D,et al.Determination of polysaccharides content in Alpinia oxyphylla[J].Chinese Journal of Modern Applied Pharmacy,2013,30(10):1070-1074.(in Chinese)
[12] 陈凌锋,蔡旭滨,檀新珠,等.太子参茎叶多糖对断奶仔猪肠道免疫功能、肠黏膜形态结构及盲肠内容物菌群的影响[J].动物营养学报,2017,29(3):1012-1020.CHEN L F,CAI X B,TAN X Z,et al.Effects of Radix pseudostellariae stem and leaf polysaccharide on intestinal immune function,intestinal mucosal morphology and cecum contents flora of weaned piglets[J].Chinese Journal of Animal Nutrition,2017,29(3):1012-1020.(in Chinese)
[13] 刘钟杰,许剑琴.中兽医学[M].4版.北京:中国农业出版社,2011:459.LIU Z J,XU J Q.Chinese herbal medicine[M].4th ed.Beijing:China Agriculture Press,2011:459.(in Chinese)
[14] 陈清华.牛膝多糖对猪的营养效应和免疫调控机理研究[D].长沙:湖南农业大学,2008.CHEN Q H.Study on the mechanism of nutrition and immunomodultion of Achyranthes bidentata polysaccharide in pigs[D].Changsha:Hunan Agricultural University,2008.(in Chinese)
[15] 左涛.鱿鱼墨多糖改善化疗小鼠肠道黏膜免疫的作用及机理探讨[D].青岛:中国海洋大学,2015.ZUO T.Protective effects of squid ink polysaccharides on intestinal mucosal immunity in chemotherapeutic mice and the underlying mechanisms[D].Qingdao:Ocean University of China,2015.(in Chinese)
[16] 翟双双.肠道黏膜屏障机能及其调控研究进展[J].现代畜牧兽医,2014(7):54-58.ZHAI S S.Progress on function and regulation of the intestinal mucosal barrier[J].Modern Journal of Animal Husbandry and Veterinary Medicine,2014(7):54-58.(in Chinese)
[17] 常建星,陈双,蒋龙元,等.小肠杯状细胞在大鼠休克后肠黏膜重建中的作用[J].中华胃肠外科杂志,2005,8(6):510-512.CHANG J X,CHEN S,JIANG L Y,et al.Effect of goblet cell in rat intestine on the restitution process of the gut barrier after hemorrhagic shock[J].Chinese Journal of Gastrointestinal Surgery,2005,8(6):510-512.(in Chinese)
[18] 石玉祥,闫金坤,王雪敏.枸杞多糖对小鼠肠道上皮内淋巴细胞和杯状细胞数量、分布及对IL-2水平影响[J].食品科学,2011,32(13):318-320.SHI Y X,YAN J K,WANG X M.Effect of Chinese wolfberry (Lycium barbarum) polysaccharides on number and distribution of intraepithelial lymphocytes and goblet cells and IL-2 expression in mice[J].Food Science,2011,32(13):318-320.(in Chinese)
[19] LINDEN S K,SUTTON P,KARLSSON N G,et al.Mucins in the mucosal barrier to infection[J].Mucosal Immunol,2008,1(3):183-197.
[20] 王娜,唐雪婵.黏蛋白-2与肠黏膜屏障损伤的研究进展[J].基础医学与临床,2015,35(7):985-988.WANG N,TANG X C.Research progress of mucin-2 and intestinal mucosal barrier damage[J].Basic & Clinical Medicine,2015,35(7):985-988.(in Chinese)
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