金荞麦对脂多糖诱导小鼠小肠炎症的保护作用

doi:10.16431/j.cnki.1671-7236.2020.02.032

摘要/Abstract

摘要： 为探究金荞麦对脂多糖（LPS）诱导的小鼠小肠炎症的保护作用，试验使用雌性ICR小鼠，前期14 d饮水添加不同浓度金荞麦和阳性对照品——芦丁（Rutin），随后腹腔注射LPS（10 mg/kg）构建小肠急性炎症模型，通过观察小鼠行为变化、血常规检测、组织切片和炎症相关基因定量检测等评估效果。结果发现，腹腔注射10 mg/kg的LPS可成功诱导小鼠急性肠炎；添加金荞麦能够剂量依赖性地降低LPS诱导的肠炎小鼠体重损失。血常规检测显示，腹腔注射LPS后导致小鼠血液白细胞数（WBC）、红细胞数（RBC）、血红蛋白（HGB）、血小板数目（PLT）显著下降，添加金荞麦或芦丁能一定程度缓解由于LPS引起的血常规异常。小肠切片HE染色结果显示，腹腔注射LPS后小肠固有层中间出现大量灶性的炎性细胞浸润，腺上皮可见少量的炎性细胞浸润，肠道细胞排列紊乱，可见核分裂及核碎裂，同时黏膜充血、出血严重；添加金荞麦或芦丁能一定程度抑制炎性细胞浸润和黏膜出血。实时荧光定量PCR结果显示，腹腔注射LPS能显著促进小肠促炎性因子IL-6、TNF-α和IL-1β的表达，添加金荞麦能剂量依赖性下调TNF-α和IL-1β的表达水平。以上结果表明饮水添加金荞麦和芦丁可以有效缓解LPS诱导的小鼠小肠炎症，金荞麦浓度为12%的添加剂量在部分指标上可达到或接近灌胃100 mg/kg芦丁的效果。

关键词: 金荞麦; 脂多糖; 肠炎; 保护作用

Abstract: To explore the protective effect of Fagopyrum dibotrys on lipopolysaccharide (LPS)-induced inflammation model of the small intestine in mice,ICR mice were selected in the experiment.Different concentration of Fagopyrum dibotrys and the positive control- Rutin were added in drinking water for 14 days treatment,then intraperitoneal injection of LPS (10 mg/kg) was used to construct the model of acute enteritis.Finally,the results were evaluated by behavior changes,routine blood tests,histological sections and gene expressions of inflammation-related genes.The results showed that the intraperitoneal injection of 10 mg/kg LPS could successfully induced acute enteritis in mice.The addition of Fagopyrum dibotrys could reduce the weight loss of LPS-induced acute enteritis in a dose-dependent manner.Blood routine test showed that intraperitoneal injection of LPS caused a significant decrease in white blood count (WBC),red blood count (RBC),hemoglobin (HGB) and platelet count (PLT) in mice,and the addition of Fagopyrum dibotrys or Rutin could alleviate the blood routine abnormality caused by LPS injection to a certain extent.The hematoxylin-eosin (HE) staining showed that after intraperitoneal injection of LPS,the focal inflammatory cells were found infiltrated into the lamina propria with a large number,as well as the glandular epithelium with a small number.Also,the disorganized intestinal cells,nuclear division and nuclear fragmentation were observed.Furthermore,severely mucosal hyperemia and hemorrhage were observed.To a certain extent,the infiltration of inflammatory cells and mucosa bleeding could be suppressed by adding Fagopyrum dibotrys or Rutin in mice.RT-PCR results showed that intraperitoneal injection of LPS could significantly promote the mRNA expression level of inflammatory factors (i.e.IL-6,TNF-α and IL-1β),and the expression of TNF-α and IL-1β decreased by adding Fagopyrum dibotrys in a dose-dependent manner.The above results showed that Fagopyrum dibotrys or Rutin could effectively alleviate the inflammation of mice small intestine induced by LPS,the 12% adding of Fagopyrum dibotrys could achieve the effect of 100 mg/kg Rutin in mice somehow.

Key words: Fagopyrum dibotrys; lipopolysaccharide; enteritis; protective effect

中图分类号:

S816.5⁺1

谭娅, 甘麦邻, 范源, 王婧, 张雄, 黄波, 朱砺, 尚以顺, 史开志. 金荞麦对脂多糖诱导小鼠小肠炎症的保护作用[J]. 中国畜牧兽医, 2020, 47(2): 597-604.

TAN Ya, GAN Mailin, FAN Yuan, WANG Jing, ZHANG Xiong, HUANG Bo, ZHU Li, SHANG Yishun, SHI Kaizhi. A Protective Effects of Fagopyrum dibotrys on Lipopolysaccharide-induced Intestinal Inflammation in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 597-604.

参考文献

[1] 李蕾,孙美利,张舒媛,等.近十年金荞麦化学成分及药理活性研究进展[J].中医药导报,2015,21(4):46-48. LI L,SUN M L,ZHANG S Y,et al.Progress of chemical constituents and pharmacological activity test of Fagopyrum dibotrys[J].Guiding Journal of Traditional Chinese Medicine and Pharmacy,2015,21(4):46-48.(in Chinese)
[2] 严晶,袁嘉嘉,刘丽娜,等.金荞麦药理作用及临床应用研究进展[J].山东中医杂志,2017,36(7):621-624. YAN J,YUAN J J,LIU L N,et al.Research progress in pharmacological action and clinical application of Fagopyrum tataricum[J].Shandong Journal of Traditional Chinese Medicine,2017,36(7):621-624.(in Chinese)
[3] GE F,ZHU S,LIU L,et al.Anti-inflammatory effects of Fagopyrum cymosum administered as a potential drug for ulcerative colitis[J].Experimental and Therapeutic Medicine,2017,14(5):4745-4754.
[4] 盛华刚,朱立俏,林桂涛.金荞麦的化学成分与药理作用研究进展[J].西北药学杂志,2011,26(2):156. SHENG H G,ZHU L Q,LIN G T.Research progress of chemical constituents and pharmacological action of buckwheat[J].Northwest Pharmaceutical Journal,2011,26(2):156.(in Chinese)
[5] LIU N,ZELLER F J,CHEN Q F.The flavonoid content in leaves and inflorescences of the wild perennial Fagopyrum cymosum complex[J].Genetic Resources and Crop Evolution,2013,60(3):825-838.
[6] 唐宇,贾洪锋,孙俊秀,等.金荞麦有效成分及含量变化研究[J].湖北农业科学,2014,53(3):672-675. TANG Y,JIA H F,SUN J X,et al.Studies on the active ingredients and its content change of Fagopyrum cymosum[J].Hubei Agricultural Sciences,2014,53(3):672-675.(in Chinese)
[7] 方福平,夏先林,顾明,等.复方金荞麦添加剂对瘦肉型猪的增重与饲料转化效果[J].贵州农业科学,2014,42(4):162-165. FANG F P,XIA X L,GU M,et al.Effects of compound buckwheat feed additives on weight gain and feed conversion of lean hogs[J].Guizhou Agricultural Sciences,2014,42(4):162-165.(in Chinese)
[8] 谭露霖,张定红,张洁,等.金荞麦对肉仔鸡生长性能、免疫功能和肠道形态结构的影响[J].中国畜牧兽医,2017,44(12):3505-3511. TAN L L,ZHANG D H,ZHANG J,et al.Effect of Fagopyrum dibotrys(D.Don) hara on growth performance,immune function and intestinal structure of broilers[J].China Animal Husbandry & Veterinary Medicine,2017,44(12):3505-3511.(in Chinese)
[9] 王航,汤承,岳华,等.金荞麦超微粉对小鼠免疫功能的影响[J].中国畜牧兽医,2013,40(9):165-168. WANG H,TANG C,YUE H,et al.Study on the effect of Fagopyrum dibotorys superfines powder on immune function of mice[J].China Animal Husbandry & Veterinary Medicine,2013,40(9):165-168.(in Chinese)
[10] 胡逸枫,梁夏瑜,王航,等.金荞麦超微粉对肉鸡免疫功能、生长发育的影响[J].中国畜牧兽医,2017,44(11):3195-3200. HU Y F,LIANG X Y,WANG H,et al.Effect of Fagopyrum dibotryis superfine powder on the immune function and the growth and development of broilers[J].China Animal Husbandry & Veterinary Medicine,2017,44(11):3195-3200.(in Chinese)
[11] 胡宇声.苦参露对脂多糖诱导大鼠小肠炎症的保护作用及其机制研究[D].北京:中国农业大学,2017. HU Y S.Protective effect and mechanisms of Kushenlu on lipopolysaccharide-induced small intestinal inflammation in rats[D].Beijing:China Agricultural University,2017.(in Chinese)
[12] 葛飞.金荞麦对溃疡性结肠炎的抗炎作用及机制研究[D].南京:南京中医药大学,2016. GE F.Anti-inflammatory effect of ethanol extracts of Fagopyrum cymosum(Trev.) on ulcerative colitis models[D].Nanjing:Nanjing University of Chinese Medicine,2016.(in Chinese)
[13] 艾蓉,薛红,罗文菊,等.黔金荞麦1号饲喂猪、鸡生产试验[J].贵州畜牧兽医,2017,41(4):5-7. AI R,XUE H,LUO W J,et al.The experiment of feeding pigs and chickens with Fagopyrum dibotrys(D.Don) No.1[J].Guizhou Journal of Animal Husbandry & Veterinary Medicine,2017,41(4):5-7.(in Chinese)
[14] 宋必卫,马传庚,田薇,等.芦丁的镇痛作用[J].安徽医科大学学报,1995,30(3):177-179. SONG B W,MA C G,TIAN W,et al.Experimental studies on the analgesia of Rutin[J].Acta University Medicinal Anhui,1995,30(3):177-179.(in Chinese)
[15] 丁琦.IL-17在脂多糖诱导的急性肺损伤中的作用及机制研究[D].苏州:苏州大学,2017. DING Q.Role and mechanism of IL-17 in LPS-induced acute lung injury[D].Suzhou:Suzhou University,2017.(in Chinese)
[16] 王月华.蓝靛果花色苷提取物对脂多糖诱导肝炎的抑制作用机制研究[D].沈阳:沈阳农业大学,2017. WANG Y H.Study on the inhibitory mechanism and effects of lonicera caerulea berry anthocyanin extracts on hepatitis induced by lipopolysaccharide[D].Shenyang:Shenyang Agricultural University,2017.(in Chinese)
[17] 翟亚南,王晶晶,李梦,等.D-氨基半乳糖联合脂多糖诱导小鼠慢性肝损伤模型的建立[J].中国比较医学杂志,2014,24(5):62-65. ZHAI Y N,WANG J J,LI M,et al.Chronic hepatic injury modeling in mice induced by D-galactosamine and lipopolysaccharide combination[J].Chinese Journal of Comparative Medicine,2014,24(5):62-65.(in Chinese)
[18] 覃文慧,杨柯,邓家刚,等.不同诱导剂致慢性阻塞性肺疾病模型小鼠氧化应激损伤的实验研究[J].世界科学技术-中医药现代化,2014,16(1):93-97. QIN W H,YANG K,DENG J G,et al.Experimental observation of lung oxidative stress injury in mice model of chronic obstructive pulmonary disease induced by different inducers[J].Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology,2014,16(1):93-97.(in Chinese)
[19] NKPAA K W,ONYESO G I.Rutin attenuates neurobehavioral deficits,oxidative stress,neuro-inflammation and apoptosis in fluoride treated rats[J].Neuroscience Letters,2018,682:92-99.
[20] LEE W,KU S K,BAE J S.Barrier protective effects of rutin in LPS-induced inflammation in vitro and in vivo[J].Food and Chemical Toxicology,2012,50(9):3048-3055.
[21] 卢携弟,吕艳琼.血常规的临床检验分析[J].中国医药指南,2013,24:116-117. LU X D,LYU Y Q.Clinical analysis of blood routine test[J].Chinese Medical Guidelines,2013,24:116-117.(in Chinese)
[22] 李海波.血常规检验中各项指标的意义[J].世界最新医学信息文摘,2016,16(42):110-111. LI H B.Significance of various indexes in blood routine test[J].World Latest Medical Information Abstract,2016,16(42):110-111.(in Chinese)
[23] SHANKAR E,VYKHOVANETS E V,VYKHOVANETS O V,et al.High-fat diet activates pro-inflammatory response in the prostate through association of Stat-3 and NF-κB[J].Prostate,2012,72(3):233-243.
[24] NI L J,WANG N N,ZHANG L G,et al.Evaluation of the effects of active fractions of Chinese medicine formulas on IL-1β,IL-6,and TNF-α release from ANA-1 murine macrophages[J].Journal of Ethnopharmacology,2016,179:420-431.
[25] LIU L,CAI X,YAN J,et al.In vivo and in vitro antinociceptive effect of Fagopyrum cymosum(Trev.) meisn extracts:A possible action by recovering intestinal barrier dysfunction[J].Evidence-Based Complementray and Alternative Medicine,2012,2012(3):1-13.
[26] 李红丽,文丹丹,周美亮,等.金荞麦抑瘤活性成份提取及作用机制研究进展[J].中国临床药理学与治疗学,2019,24(7):833-840. LI H L,WEN D D,ZHOU M L,et al.Advances in extraction and mechanism of antitumor active components from Fagopyrum dibotrys[J].Chinese Journal of Clinical Pharmacology and Therapeutics,2019,24(7):833-840.(in Chinese)
[27] ELSAYED H E,EBRAHIM H Y,MOHYELDIN M M,et al.Rutin as a novel c-Met inhibitory lead for the control of triple negative breast malignancies[J].Nutrition and Cancer,2017,69(8):1256-1271.