博落回叶提取物对小鼠溃疡性结肠炎的影响

doi:10.16431/j.cnki.1671-7236.2023.11.034

摘要/Abstract

摘要： 【目的】研究博落回叶提取物（Macleaya cordata leaf extract，MCLE）对葡聚糖硫酸钠（DSS）诱导的溃疡性结肠炎（UC）小鼠的保护作用，为博落回叶的资源利用和药物开发提供理论依据。【方法】选择60只C57BL/6小鼠，随机分成6组，即对照组（CON）组、DSS模型组、美沙拉嗪（MES）阳性对照组（200 mg/kg）及MCLE高（MCLE-H，450 mg/kg）、中（MCLE-M，150 mg/kg）、低（MCLE-L，50 mg/kg）剂量组，试验期14 d。通过小鼠的一般症状、体重变化、疾病活动指数（DAI）、脾脏指数、结肠长度及组织病理学等指标评价MCLE对DSS诱导的小鼠结肠炎模型的治疗效果。使用ELISA和实时荧光定量PCR分别检测血清和结肠组织中白细胞介素-1β（IL-1β）、IL-6、肿瘤坏死因子α（TNF-α）的表达水平；比色法测定结肠组织中髓过氧化物酶（MPO）和一氧化氮（NO）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）、超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量；通过分析16S rRNA基因序列确定MCLE对肠道菌群的影响；采用气相色谱法测定结肠内容物中短链脂肪酸（SCFAs）的含量。【结果】MCLE处理组均极显著抑制了UC小鼠的体重降低和结肠长度的缩短（P<0.01），并显著降低了小鼠DAI、脾脏指数及结肠组织病理学评分（P<0.05）；MCLE 显著下调了结肠和血清中 IL-1β、IL-6和TNF-α的表达（P<0.05），降低了结肠组织中 MPO、NO和MDA 的含量（P<0.05），增加了GSH-Px、SOD活性及CAT含量（P<0.05）；与DSS组相比，拟杆菌属、另枝菌属、阿克曼氏菌属等有益菌属在MCLE给药组中丰度占比增加，病原菌如脱硫弧菌属等占比减少。此外，MCLE治疗还显著提高了UC小鼠肠内容物中SCFAs（乙酸、丙酸、丁酸）的水平（P<0.05）。【结论】MCLE可能是通过调节炎症因子水平、减轻氧化应激影响、维持肠道菌群稳态和恢复短链脂肪酸的产生发挥抗结肠炎作用。

关键词: 博落回叶提取物; 小鼠; 溃疡性结肠炎; 炎症因子; 肠道菌群; 短链脂肪酸

Abstract: 【Objective】 The purpose of this experiment was to study the protection of Macleaya cordata leaf extract (MCLE) on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) mice, and provide a theoretical basis for resource utilization and drug development of Macleaya cordata leaves.【Method】 Sixty C57BL/6 mice were randomly divided into 6 groups:Control group (CON), dextran sodium sulfate (DSS) model group, mesalazine (MES) positive control group (200 mg/kg), and MCLE high (MCLE-H, 450 mg/kg), medium (MCLE-M, 150 mg/kg) and low (MCLE-L, 50 mg/kg) dose groups.The test lasted for 14 days.The therapeutic effect of MCLE on DSS-induced colitis in mice was evaluated by general signs, body weight change, disease activity index (DAI), spleen index, colon length and histopathology.The expression levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) in serum and colon tissues were detected by ELISA and Real-time quantitative PCR, respectively.The contents of myeloperoxidase (MPO), nitric oxide (NO), catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and malondialdehyde (MDA) in colonic tissue were determined by colorimetry.The effect of MCLE on intestinal flora was determined by analyzing 16S rRNA gene sequence.The content of short chain fatty acids (SCFAs) in colonic contents was determined by gas chromatography.【Result】 MCLE treatment significantly inhibited the weight loss and colon length shortening of UC mice (P<0.01), and significantly reduced DAI, spleen index and colon histopathological score (P<0.05).Moreover, MCLE significantly down-regulated the expression of IL-1β, IL-6 and TNF-α in colon and serum (P<0.05), decreased the content of MPO, NO and MDA in colon tissue (P<0.05), and increased the content of GSH-Px, SOD and CAT (P<0.05).At the same time, compared with DSS group, the abundance of beneficial bacteria such as Bacteroides, Alistipes and Akkermansia increased in MCLE administration group, and the proportion of pathogenic bacteria such as Desulfovibrio decreased.In addition, MCLE treatment also significantly increased the levels of SCFAs (acetic acid, propionic acid, butyric acid) in intestinal contents of UC mice (P<0.05).【Conclusion】 MCLE might play an anti-colitis role by regulating the level of inflammatory factors, reducing the effects of oxidative stress, maintaining intestinal flora homeostasis and restoring the production of SCFAs.

Key words: Macleaya cordata leaf extract; mice; ulcerative colitis; inflammatory factors; intestinal flora; short chain fatty acids

中图分类号:

S859.3

马晓兰, 董朕, 岳美杉, 常冠宇, 曾建国. 博落回叶提取物对小鼠溃疡性结肠炎的影响[J]. 中国畜牧兽医, 2023, 50(11): 4664-4680.

MA Xiaolan, DONG Zhen, YUE Meishan, CHANG Guanyu, ZENG Jianguo. Effect of Macleaya cordata Leaf Extract on Ulcerative Colitis in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4664-4680.

参考文献

[1] TEIRLYNCK E, GUSSEM M D E, DEWULF J, et al.Morphometric evaluation of "dysbacteriosis" in broilers[J].Avian Pathology, 2011, 40(2):139-144.
[2] 张金武, 魏紫, 冀禹彤, 等.A型产气荚膜梭菌诱导黄羽肉鸡肠道氧化应激和炎性损伤研究[J].中国畜牧兽医, 2023, 50(5):2069-2081. ZHANG J W, WEI Z, JI Y T, et al.Study on oxidative stress and inflammatory injury in intestinal tract of Yellow-feathered chickens induced by Clostridium perfringens type A[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(5):2069-2081.(in Chinese)
[3] 杨志, 余佳欣, 杨林富, 等.四逆汤加减对仔猪腹泻治疗效果研究[J].中国畜牧兽医, 2023, 50(4):1707-1717. YANG Z, YU J X, YANG L F, et al.Study on the therapeutic effect of modified Sini decoction on piglet diarrhea[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(4):1707-1717.(in Chinese)
[4] 常丽云, 刘志勇, 秦建华, 等.河北省唐山市奶犊牛病毒性腹泻流行病学调查[J].中国畜牧兽医, 2021, 48(11):4213-4219. CHANG L Y, LIU Z Y, QIN J H, et al.Epidemiological investigation of viral diarrhea of dairy calves in Tangshan city, Hebei province[J].China Animal Husbandry & Veterinary Medicine, 2021, 48(11):4213-4219.(in Chinese)
[5] DUCATELLE R, GOOSSENS E, DE MEYER F, et al.Biomarkers for monitoring intestinal health in poultry:Present status and future perspectives[J].Veterinary Research, 2018, 49(1):43-51.
[6] SONG L, ZHANG Y, ZHU C, et al.Hydrogen-rich water partially alleviate inflammation, oxidative stress and intestinal flora dysbiosis in DSS-induced chronic ulcerative colitis mice[J].Advances in Medical Sciences, 2022, 67(1):29-38.
[7] GRESSE R, CHAUCHEYRAS-DURAND F, FLEURY M A, et al.Gut microbiota dysbiosis in postweaning piglets:Understanding the keys to health[J].Trends in Microbiology, 2017, 25(10):851-873.
[8] LE BERRE C, ANANTHAKRISHNAN A N, DANESE S, et al.Ulcerative colitis and Crohn's disease have similar burden and goals for treatment[J].Clinical Gastroenterology and Hepatology, 2020, 18(1):14-23.
[9] HOFACRE C L, SMITH J A, MATHIS G F.An optimist's view on limiting necrotic enteritis and maintaining broiler gut health and performance in today's marketing, food safety, and regulatory climate[J].Poultry Science, 2018, 97(6):1929-1933.
[10] ASLAM N, LO S W, SIKAFI R, et al.A review of the therapeutic management of ulcerative colitis[J].Therapeutic Advances in Gastroenterology, 2022, 15:17562848221138160.
[11] DASSOU G H, JÉRONIME M A S O, ADOMOU A C, et al.Plant and natural product based homemade remedies for veterinary uses by the Peul community in Benin[J].Journal of Ethnopharmacology, 2020, 261:113107.
[12] 黄鹏, 刘金凤, 徐敏, 等.基于ITS2序列的博落回属植物鉴定研究[J].中国现代中药, 2017, 19(10):1367-1370. HUANG P, LIU J F, XU M, et al.Identification of Macleaya plants based on ITS2 sequence[J].Modern Chinese Medicine, 2017, 19(10):1367-1370.(in Chinese)
[13] YU X, GAO X, ZHU Z, et al.Alkaloids from the tribe Bocconieae (Papaveraceae):A chemical and biological review[J].Molecules, 2014, 19(9):13042-13060.
[14] LIU Y, WANG Q, LIU H, et al.Effects of dietary Bopu powder supplementation on intestinal development and microbiota in broiler chickens[J].Frontiers in Microbiology, 2022, 13:1019130.
[15] LIU H, LIN Q, LIU X, et al.Effects of dietary Bopu powder supplementation on serum antioxidant capacity, egg quality, and intestinal microbiota of laying hens[J].Frontiers in Physiology, 2022, 13:902784.
[16] LIU G, GUAN G P, FANG J, et al. Macleaya cordata extract decreased diarrhea score and enhanced intestinal barrier function in growing piglets[J].Biomed Research International, 2016, 2016:1069585.
[17] HUANG P, ZHANG Y, XIAO K, et al.The chicken gut metagenome and the modulatory effects of plant-derived benzylisoquinoline alkaloids[J]. Microbiome, 2018, 6(1):211.
[18] 孙晓蛟, 金海峰, 闫研, 等.不同抗生素替代品组合对断奶仔猪生长性能、腹泻及抗氧化能力的影响[J].中国畜牧兽医, 2020, 47(2):460-468. SUN X J, JIN H F, YAN Y, et al.Effects of different combinations of antibiotic substitute on growth performance, diarrhea rate and antioxidant indexes of weaned piglets[J].China Animal Husbandry & Veterinary Medicine, 2020, 47(2):460-468.(in Chinese)
[19] LIU X B, LIU Y S, HUANG P, et al.The genome of medicinal plant Macleaya cordata provides new insights into benzylisoquinoline alkaloids metabolism[J]. Molecular Plant, 2017, 10(7):975-989.
[20] WIRTZ S, POPP V, KINDERMANN M, et al.Chemically induced mouse models of acute and chronic intestinal inflammation[J].Nature Protocols, 2017, 12(7):1295-1309.
[21] DIELEMAN L A, PALMEN M, AKOL H, et al.Chronic experimental colitis induced by dextran sulphate sodium (DSS) is characterized by Th1 and Th2 cytokines[J].Clinical & Experimental Immunology, 1998, 114(3):385-391.
[22] 黄嘉璐.基于NF-κB/GSDMD通路探究博普总碱对脂多糖致肠炎的抗炎机制研究[D].太谷:山西农业大学, 2022. HUANG J L.Study on the anti-inflammatory mechanism of bopp base on lipopolysaccharide-induced enteritis based on NF-κB/GSDMD pathway[D].Taigu:Shanxi Agricultural University, 2022.(in Chinese)
[23] ORDÁS I, ECKMANN L, TALAMINI M, et al.Ulcerative colitis[J].Lancet, 2012, 380(9853):1606-1619.
[24] LI H, FAN C, LU H, et al.Protective role of berberine on ulcerative colitis through modulating enteric glial cells-intestinal epithelial cells-immune cells interactions[J]. Acta Pharmaceutica Sinica B, 2020, 10(3):447-461.
[25] DHANISHA S S, GURUVAYOORAPPAN C, DRISHYA S, et al.Mucins:Structural diversity, biosynthesis, its role in pathogenesis and as possible therapeutic targets[J]. Critical Reviews in Oncology/Hematology, 2018, 122:98-122.
[26] REINDERS C I, JONKERS D, JANSSON E Å, et al.Rectal nitric oxide and fecal calprotectin in inflammatory bowel disease[J]. Scandinavian Journal of Gastroenterology, 2007, 42(10):1151-1157.
[27] MIDDLETON S J, SHORTHOUSE M, HUNTER J O.Increased nitric oxide synthesis in ulcerative colitis[J].The Lancet, 1993, 341(8843):465-466.
[28] ISCHIROPOULOS H, GOW A.Pathophysiological functions of nitric oxide-mediated protein modifications[J].Toxicology, 2005, 208(2):299-303.
[29] BUCHAN G J, BONACCI G, FAZZARI M, et al.Nitro-fatty acid formation and metabolism[J].Nitric Oxide, 2018, 79:38-44.
[30] MU K, YU S, KITTS D D.The role of nitric oxide in regulating intestinal redox status and intestinal epithelial cell functionality[J].International Journal of Molecular Sciences, 2019, 20(7):1755.
[31] SALOMON J D, QIU H, FENG D, et al.Piglet cardiopulmonary bypass induces intestinal dysbiosis and barrier dysfunction associated with systemic inflammation[J].Disease Models & Mechanisms, 2023, 16(5):dmm049742.
[32] ABD ELMAKSOUD H A, MOTAWEA M H, DESOKY A A, et al.Hydroxytyrosol alleviate intestinal inflammation, oxidative stress and apoptosis resulted in ulcerative colitis[J].Biomedicine & Pharmacotherapy, 2021, 142:112073.
[33] MOURA F A, DE ANDRADE K Q, DOS SANTOS J C F, et al.Antioxidant therapy for treatment of inflammatory bowel disease:Does it work?[J].Redox Biology, 2015, 6:617-639.
[34] DAGLI V, BALK M, YVCEL D, et al.The role of reactive oxygen metabolites in ulcerative colitis[J].Inflammatory Bowel Diseases, 1997, 3(4):260-264.
[35] LEVY A N, ALLEGRETTI J R.Insights into the role of fecal microbiota transplantation for the treatment of inflammatory bowel disease[J].Therapeutic Advances in Gastroenterology, 2019, 12:1756.
[36] LIN X, SHI Y, CAO Y, et al.Sanguinarine protects against indomethacin-induced small intestine injury in rats by regulating the Nrf2/NF-κB pathways[J].Frontiers in Pharmacology, 2022, 13:960140.
[37] LI X, WU X, WANG Q, et al.Sanguinarine ameliorates DSS induced ulcerative colitis by inhibiting NLRP3 inflammasome activation and modulating intestinal microbiota in C57BL/6 mice[J].Phytomedicine, 2022, 104:154321.
[38] WU Y, ZHAO N J, CAO Y, et al.Sanguinarine metabolism and pharmacokinetics study in vitro and in vivo[J].Journal of Veterinary Pharmacology and Therapeutics, 2020, 43(2):208-214.
[39] HU N X, CHEN M, LIU Y S, et al.Pharmacokinetics of sanguinarine, chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration[J].Journal of Veterinary Pharmacology and Therapeutics, 2019, 42(2):197-206.
[40] SHEN L X, LIU G F, SONG J S, et al.Sex differences in the pharmacokinetics and tissue residues of Macleaya cordata extracts in rats[J]. Xenobiotica, 2022, 52(1):46-53.
[41] GONÇALVES P, ARAÚJO J R, DI SANTO J P.A cross-talk between microbiota-derived short-chain fatty acids and the host mucosal immune system regulates intestinal homeostasis and inflammatory bowel disease[J]. Inflammatory Bowel Diseases, 2018, 24(3):558-572.
[42] HALFVARSON J, BRISLAWN C J, LAMENDELLA R, et al.Dynamics of the human gut microbiome in inflammatory bowel disease[J]. Nature Microbiology, 2017, 2(5):17004.
[43] DE PRETER V, ARIJS I, WINDEY K, et al.Impaired butyrate oxidation in ulcerative colitis is due to decreased butyrate uptake and a defect in the oxidation pathway[J]. Inflammatory Bowel Diseases, 2012, 18(6):1127-1136.
[44] BOESMANS L, RAMAKERS M, ARIJS I, et al.Inflammation-induced downregulation of butyrate uptake and oxidation is not caused by a reduced gene expression[J].Journal of Cellular Physiology, 2015, 230(2):418-426.
[45] WANG M X, LIN L, CHEN Y D, et al.Evodiamine has therapeutic efficacy in ulcerative colitis by increasing Lactobacillus acidophilus levels and acetate production[J].Pharmacological Research, 2020, 159:104978.
[46] CUI L, GUAN X, DING W, et al.Scutellaria baicalensis Georgi polysaccharide ameliorates DSS-induced ulcerative colitis by improving intestinal barrier function and modulating gut microbiota[J].International Journal of Biological Macromolecules, 2021, 166:1035-1045.
[47] LEE J, D'AIGLE J, ATADJA L, et al.Gut microbiota-derived short-chain fatty acids promote poststroke recovery in aged mice[J]. Circulation Research, 2020, 127(4):453-465.
[48] SUBRAMANI D, JOHANSSON M, DAHLEN G, et al.Lactobacillus and Bifidobacterium species do not secrete protease that cleaves the MUC2 mucin which organises the colon mucus[J]. Beneficial Microbes, 2010, 1(4):343-350.
[49] CHENG H, ZHANG D, WU J, et al.Atractylodes macrocephala Koidz.volatile oil relieves acute ulcerative colitis via regulating gut microbiota and gut microbiota metabolism[J]. Frontiers in Immunology, 2023, 14:1127785.
[50] HONG T, ZOU J, HE Y, et al.Bisphenol A induced hepatic steatosis by disturbing bile acid metabolism and FXR/TGR5 signaling pathways via remodeling the gut microbiota in CD-1 mice[J].Science of the Total Environment, 2023, 889:164307.
[51] LI S, WANG T, WU B, et al.Anthocyanin-containing purple potatoes ameliorate DSS-induced colitis in mice[J].The Journal of Nutritional Biochemistry, 2021, 93:108616.
[52] LI K, HAO Z, DU J, et al.Bacteroides thetaiotaomicron relieves colon inflammation by activating aryl hydrocarbon receptor and modulating CD4⁺ T cell homeostasis[J].International Immunopharmacology, 2021, 90:107183.
[53] KIM J Y, KWON Y M, KIM I S, et al.Effects of the brown seaweed Laminaria japonica supplementation on serum concentrations of IgG, triglycerides, and cholesterol, and intestinal microbiota composition in rats[J]. Frontiers in Nutrition, 2018, 5:23.
[54] DZIARSKI R, PARK S Y, KASHYAP D R, et al.Pglyrp-regulated gut microflora Prevotella falsenii, Parabacteroides distasonis and Bacteroides eggerthii enhance and Alistipes finegoldii attenuates colitis in mice[J].PLoS One, 2016, 11(1):e0146162.
[55] DIN A U, HASSAN A, ZHU Y, et al.Inhibitory effect of Bifidobacterium bifidum ATCC 29521 on colitis and its mechanism[J].The Journal of Nutritional Biochemistry, 2020, 79:108353.
[56] VERSTREKEN I, LALEMAN W, WAUTERS G, et al.Desulfovibrio desulfuricans bacteremia in an immunocompromised host with a liver graft and ulcerative colitis[J].Journal of Clinical Microbiology, 2012, 50(1):199-201.
[57] JANDHYALA S M, TALUKDAR R, SUBRAM-ANYAM C, et al.Role of the normal gut microbiota[J].World Journal of Gastroenterology, 2015, 21(29):8787.