| 438 | 1 | 17 |
| 下载次数 | 被引频次 | 阅读次数 |
目的：探究小陷胸汤对5-氟尿嘧啶(5-fluorouracil, 5-FU)介导的小鼠肠黏膜损伤的保护作用。方法：将100只SPF级雄性ICR小鼠随机分为正常对照组、模型组、小陷胸汤低、中、高剂量(8.3、16.6、33.2g/kg)组。除正常对照组外，其余各组采用5-FU(30 mg/kg, ip, 7 d)诱导肠黏膜损伤小鼠模型。小陷胸汤给药组造模同时按照设定剂量以10 mL·kg-1·d-1灌胃，正常对照组和模型组给予等量蒸馏水，连续处理7 d。采用苏木素-伊红染色法(HE)观察小肠的病理形态学变化并测定肠绒毛高度/隐窝深度(VH:CD)比值；采用阿利新蓝染色法(AB)测定小肠黏膜杯状细胞数量；采用分光光度法检测血清内毒素(ET)、D-乳酸(D-LA)及小肠组织二胺氧化酶(DAO)水平；采用酶联免疫吸附法(ELISA)检测血清肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平；采用实时荧光定量聚合酶链式反应法(RT-qPCR)检测小肠组织IL-6、信号转导和转录激活因子3(STAT3)和表皮生长因子受体(EGFR) mRNA表达；采用Western blot法检测小肠组织IL-6、STAT3及EGFR蛋白表达。结果：与正常对照组相比，模型组小鼠体质量显著降低，腹泻评分、粪便质量、粪便含水量及小肠推进率明显升高(P<0.01)，小肠黏膜VH:CD比值和杯状细胞数量降低(P<0.01)，血清ET和D-LA水平升高，小肠组织DAO水平降低(P<0.05)，脾指数和胸腺指数降低(P<0.01)，血清IL-6、TNF-α水平升高(P<0.05)，小肠组织IL-6、STAT3 mRNA表达升高，EGFR mRNA表达降低(P<0.01)；小肠组织IL-6、STAT3蛋白表达升高，EGFR蛋白表达降低(P<0.01)。与模型组相比，小陷胸汤能够明显改善上述指标(P<0.05)。结论：小陷胸汤对5-FU介导的肠黏膜损伤具有保护作用，其作用机制可能与小陷胸汤激活EGFR信号转导，下调IL-6/STAT3信号通路，减轻5-FU诱导的肠黏膜炎症反应，改善免疫功能有关。
Abstract：Objective: To investigate how Xiaoxianxiong decoction can protect against intestinal damage caused by 5-fluoroura-cil(5-FU) in mice.Methods: One hundred male ICR breed mice with SPF-grade were randomised into the normal control group, the model group and the low, medium and high dose(8.3, 16.6 and 33.2 g/kg) groups of Xiaoxianxiong decoction. Intestinal mu-cosal injury was induced in all groups, except the control group, by administering 5-FU(30 mg/kg, ip, 7 d). While modeling was ongoing, the mice in the Xiaoxianxiong decoction group were administered 10 mL·kg-1·d-1 via gavage, following the prescribed dos-age. The control and model groups were administered equal volume of distilled water and treated continuously for 7 d. Histopatho-logical alterations in the small intestine were assessed utilizing hematoxylin-eosin staining(HE), with the villus height to crypt depth ratio(VH:CD) subsequently determined. The number of goblet cells in the small intestinal mucosa was determined by Al-cian blue staining(AB). The concentrations of endotoxin(ET) and D-lactic acid(D-LA) in serum were measured using spectro-photometry, along with diamine oxidase(DAO) levels in the small intestine. The levels of interleukin-6(IL-6) and tumor necro-sis factor-α(TNF-α) in serum were detected using enzyme-linked immunosorbent assay(ELISA). The mRNA expression of IL-6, signal transducer and activator of transcription 3(STAT3), and epidermal growth factor receptor(EGFR) was detected us-ing reverse transcription quantitative polymerase chain reaction(RT-qPCR). The IL-6, STAT3 and EGFR protein were expres-sion in small intestinal tissues were detected by the Western blot method. Results: Compared to the control group, the model group of mice presented a significant reduction in body mass and an increase in scores for diarrhoea, faecal mass, faecal water con-tent, and small intestinal propulsion rates.(P<0.01). The VH:CD ratio, as well as the number of goblet cells in the small intesti-nal mucosa, exhibited a noteworthy decrease(P<0.01). The levels of ET and D-LA in the serum significantly augmented, while the level of DAO in the small intestine was significantly decreased(P<0.05). Significant reductions were observed in the indices for the spleen and thymus(P<0.01). A significant increase was observed in the serum levels of IL-6 and TNFα(P<0.05). IL-6and STAT3 mRNA expression were significantly up-regulated, whilst EGFR mRNA expression was down-regulated in small bowel tissues(P<0.01). The expression of IL-6 and STAT3 proteins was found to be significantly higher, while that of EGFR proteins was significantly lower, in small intestinal tissues(P<0.01). Compared to the model group, Xiaoxianxiong decoction led to a significant improvement in the aforementioned indices(P<0.05).Conclusion: Xiaoxianxiong decoction can safeguard the in-testinal mucosa against damage caused by 5-FU. The mechanism of action may involve the activation of EGFR signalling, down-regulation of the IL-6/STAT3 signalling pathway, reduction of 5-FU-induced inflammation of the intestinal mucosa, and improvement of immune function.
1晁艳红，杨广建，齐丽娟，等.5-氟尿嘧啶及其衍生物抗肿瘤作用的研究进展[J].癌症进展，2019,17(1):9-12.Chao YH,Yang GJ,Qi LJ,et al.Progress in the study of the antitumour effects of 5-fluorouracil and its derivatives[J].Oncol Prog,2019,17(1):9-12.
2 Yin L,Huang G,Khan I,et al.Poria cocos polysaccha-rides exert prebiotic function to attenuate the adverse ef-fects and improve the therapeutic outcome of 5-FU in Apc Min/+mice[J].Chin Med,2022,17(1):116.
3 Sauruk Da Silva K,Carla Da Silveira B,Bueno L R,et al.Beneficial effects of polysaccharides on the epithelial barrier function in intestinal mucositis[J].Front physiol,2021,12:714846.
4 Dahlgren D,Sjoblom M,Hellstrom PM,et al.Chemotherapeutics-induced intestinal mucositis:patho-physiology and potential treatment strategies[J].Front Pharmacol,2021,12:681417.
5 Bowen JM,Gibson RJ,Coller JK,et al.Systematic re-view of agents for the management of cancer treatment-related gastrointestinal mucositis and clinical practice guidelines[J].Support Care Cancer,2019,27(10):4011-4022.
6 Barbosa S,Oliveira M,Ribeiro SB,et al.The beneficial ef-fects of Lacticaseibacillus casei on the small intestine and colon of Swiss mice against the deleterious effects of 5-fluo-rouracil[J].Front Immunol,2022,13:954885.
7 Ying M,Yu Q,Zheng B,et al.Cultured cordyceps sinen-sis polysaccharides attenuate cyclophosphamide-induced intestinal barrier injury in mice[J].J Funct Foods,2019,62:103523.
8徐露，王娴，王宣璎，等.姜黄素介导IL-6/STAT3信号通路修复结肠癌5-FU化疗引起的肠黏膜损伤[J].中国中药杂志，2021,46(3):670-677.Xu L,Wang X,Wang XY,et al.Curcumin mediates l L-6/STAT3 signaling pathway to repair intestinal mucosalinjury induced by 5-FU chemotherapy for colon cancer[J].China J of Chin Materia Medica,2021,46(3):670-677.
9丁芮，王翔，周鹏，等.小陷胸汤中抗肿瘤作用有效成分及其机制研究进展[J].中国实验方剂学杂志，2020,26(22):186-192.Ding R,Wang X,Zhou P,et al.Research Progress on Antitumor Active Components and Mechanism of Xiao Xianxiongtang[J].Chin J Experiment Tradit Med Form,2020,26(22):186-192.
10高世勇，李婉秋，王帅，等.基于网络药理学的小陷胸汤治疗大肠癌的机制研究[J].中草药，2022,53(3):773-782.Gao SY,Li WQ,Wang S,et al.Mechanism of Xiaoxianxiong Decoction on treating colorectal cancer based onnetwork pharmacology[J].Chinese Traditional and Herbal Drugs,2022,53(03):773-782.
11贾仲琦，李竹英.麻杏石甘汤合小陷胸汤加减治疗痰热壅肺证重症肺炎患者的临床疗效及对痰液排出、氧合指数的影响[J].世界中西医结合杂志，2023,18(02):373-376.Jia ZQ,Li ZY.Clinical Effect of Modified Maxing Shigan Decoction and Xiaoxianxiong Decoction in the Treatment of Severe Pneumonia with the Syndrome of Phlegm-Heat Obstructing the Lung and lmpact on Sputum Excretion and Oxygenation Index[J].World JIntegr Tradit West Med,2023,18(02):373-376.
12 Zhong X,Surh Y,Do S,et al.Baicalein Inhibits Dex-tran Sulfate Sodium-induced Mouse Colitis[J].J Cancer Prev,2019,24(2):129-138.
13陈琴，张志云，石西南，等.基于IL-6/STAT3信号通路探讨mi R-34a-5p过表达对溃疡性结肠炎的影响及小檗碱的干预作用[J].山东医药，2024,64(1):1-5.Chen Q,Zhang ZY,Shi XN,et al.Effect of mi R-34a-5p overexpression on ulcerative colitis and the intervention of berberine based on l L-6/STAT3 signaling pathway[J].Shandong Med J,2024,64(1):1-5.
14 Chen H,Zhang F,Li R,et al.Berberine regulates fecal metabolites to ameliorate 5-fluorouracil induced intestinal mucositis through modulating gut microbiota[J].Biomed Pharmacother,2020,124:109829.
15 Sakai H,Tabata S,Kimura M,et al.Active Ingredients of Hange-shashin-to,Baicalelin and 6-Gingerol,Inhibit5-Fluorouracil-Induced Upregulation of CXCL1 in the Colon to Attenuate Diarrhea Development[J].Biol Pharm Bull,2017,40(12):2134-2139.
16 Kamran S,Sinniah A,Chik Z,et al.Diosmetin Exerts Synergistic Effects in Combination with 5-Fluorouracil in Colorectal Cancer Cells[J].Biomedicines,2022,10(3):531.
17肖刘成，曾思宇，陈伟业，等.基于16S r RNA测序研究小陷胸汤对动脉粥样硬化小鼠肠道菌群结构的影响[J].中华中医药杂志，2023,38(2):848-852.Xiao LC,Zeng SY,Chen WY,et al.Effects of Xiaoxianxiong Decoction on intestinal flora structure of atherosclerosis mice based on 16S r RNA sequencing[J].China J Tradit Chin Med Pharm,2023,38(2):848-852.
18 Wu L,Xi Y,Yan M,et al.Berberine-Based Carbon Quantum Dots Improve Intestinal Barrier Injury and Al-leviate Oxidative Stress in C57BL/6 Mice with5-Fluorouracil-Induced Intestinal Mucositis by Enhanc-ing Gut-Derived Short-Chain Fatty Acids Contents[J].Molecules,2023,28(5):2148.
19黄继汉，黄晓晖，陈志扬，等.药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学，2004,9(9):1069-1072.Huang JH,Huang XH,Chen ZY,et al.Dose conversion among different animals and healthy volunteers in pharmacological study[J].Chin J Clin Pharm Ther,2004,9(9):1069-1072.
20 Kurita A,Kado S,Kaneda N,et al.Modified irinotecan hydrochloride (CPT-11) administration schedule im-proves induction of delayed-onset diarrhea in rats[J].Cancer Chemother Pharmacol,2000,46(3):211-220.
21 Han B,Zheng R,Zeng H,et al.Cancer incidence and mortality in China,2022[J].J Nat Cancer Cent,2024,4(1):47-53.
22 Sougiannis AT,Vander Veen BN,Davis JM,et al.Un-derstanding chemotherapy-induced intestinal mucositis and strategies to improve gut resilience[J].Am J Physiol Gastrointest Liver Physiol,2021,320(5):G712-G719.
23姜彧，孙磊涛，严雯婷，等.中药方剂防治化疗消化道不良反应的研究进展[J].中华中医药杂志，2022,37(11):6305-6308.Jiang Y,Sun LT,Yan WT,et al.Research progress on prevention and treatment of gastrointestinal adverse reactions of chemotherapy with Chinese medicine prescriptions[J].Chin J Tradit Chin Med Pharm,2022,37(11):6305-6308.
24蒋梅，周岱翰.国医大师周岱翰教授从升降学说辨治肿瘤化疗致消化道不良反应经验[J].世界中西医结合杂志，2019,14(4):486-489.Jiang M,Zhou DH.Professor Zhou Daihan's experiences in treatment of digestive adverse reaction induced by chemotherapy in cancer on the base of the ascending anddescending theory[J].World J Integr Tradit West Med,2019,14(4):486-489.
25王思佳，芦殿荣，朱世杰，等.“半夏-黄连”药对防治化疗相关性腹泻的网络药理学研究[J].中国中医急症，2023,32(4):575-580.Wang SJ,Lu DR,Zhu SJ,et al.Network Pharmacologic Study on the Mechanism of"Banxia-Huanglian"in the Treatment of Chemotherapy-induced Diarrhea[J].J Emerg Tradit Chin Med,2023,32(4):575-580.
26 Zhao H,Gong J,Li L,et al.Vitamin E relieves chronic obstructive pulmonary disease by inhibiting COX2-medi-ated p-STAT3 nuclear translocation through the EGFR/MAPK signaling pathway[J].Lab Invest,2022,102(3):272-280.
27毛甜甜，白衡，贾宁.芒柄花素对免疫抑制小鼠小肠黏膜免疫功能的影响[J].动物营养学报，2022,34(4):2712-2720.Mao TT,Bai H,Jia N.Effects of Formonetin on Mucosa Immunity Function in Small Intestinal of Immunosuppressive Mice[J].Chin J Anim Nut,2022,34(4):2712-2720.
28尉浩斌，张小丹，李苏宜.肠屏障功能损伤机制及其临床检测方法研究现状[J].肿瘤代谢与营养电子杂志，2020,7(4):407-414.Yu HB,Zhang XD,Li SY.Research status of the mechanism of intestinal barrier function injury and its clinical detection methods[J].Electron J Metabol Nutrit Cancer,2020,7(4):407-414.
29张永龙，马唯刚，钱星羽，等.脾虚证实验动物模型构建及评价方法的研究述评[J/OL].中国实验动物学报（2024-03-18)[2024-05-21].http：//kns.cnki.net/kcms/detail/11.2986.Q.20240316.1506.006.html.Zhang YL,Ma WG,Qian XY,et al.Construction of experimental animal models and evaluation of spleen deficiency syndrome:A review[J/OL].Acta Laboratorium Animalis Scientia Sinica (2024-03-18)[2024-05-21].http：//kns.cnki.net/kcms/detail/11.2986.Q.20240316.1506.006.html.
30赵龙举，高建杭，高慧香，等.龙脑樟叶粉对攸县麻鸭繁殖性能、血清免疫、抗氧化和肠道健康指标的影响[J].黑龙江畜牧兽医，2024,(1):80-89.Zhao LJ,Gao JH,Gao HX,et al.Effects of Borneol Cinnamomum camphora leaf powder on reproductive performance,serum immunity,antioxidant and intestinal health indexes of Youxian partridge ducks[J].Heilongjiang Anim Sci Veter Med,2024,(1):80-89.
31 Wu X,Wei S,Chen M,et al.P2RY13 Exacerbates In-testinal inflammation by damaging the intestinal mucosal barrier via activating IL-6/STAT3 Pathway[J].Int J Bi-ol Sci,2022,18(13):5056-5069.
32 Ruder B,Atreya R,Becker C.Tumour necrosis factor alpha in intestinal homeostasis and gut related diseases[J].Int J Mol Sci,2019,20(8):1887.
33 Wang Y,Duan X,Liu X,et al.Rho kinase blockade ameliorates DSS-induced ulcerative colitis in mice through dual inhibition of the NF-κB and IL-6/STAT3pathways[J].Inflammation,2020.
34 Chen XQ,Lv XY,Liu SJ.Baitouweng decoction allevi-ates dextran sulfate sodium-induced ulcerative colitis by regulating intestinal microbiota and the IL-6/STAT3 sig-naling pathway[J].J Ethnopharmacol,2021,265:113357.
35李哲，李娜，崔晓燕，等.基于网络药理学和实验验证探讨陈皮藿香汤治疗腹泻的作用机制[J].中国实验方剂学杂志，2022,28(23):79-86.Li Z,Li N,Cui XY,et al.Mechanism of Chenpi Huoxiangtang Against Diarrhea:Based on Network Pharmacology and Experimental Verification[J].Chin JExperiment Tradit Med Form,2022,28(23):79-86.
36 Guan X,Hou Y,Sharma S,et al.Methyltransferase smyd2 modulates therapeutic effects of epidermal growth factor in inflammatory bowel disease[J].Inflamm Bowel Dis,2022,28(Sup1):S50-S51.
37王雪，蒙臣，曾文，等.表皮生长因子受体信号通路影响紧密连接的作用与机制研究进展[J].中国医药导报，2020,17(15):37-40.Wang X,Meng C,Zeng W,et al.Research progress on the effects and mechanisms of epithelial growth factor receptor on tight junction[J].China Med Her,2020,17(15):37-40.
基本信息:
DOI：10.13210/j.cnki.jhmu.20240524.002
中图分类号:R285.5
引用信息:
[1]秦美洁,叶倩倩,丁芮,等.小陷胸汤对5-氟尿嘧啶介导的小鼠肠黏膜损伤的保护作用研究[J].海南医学院学报,2024,30(19):1441-1450.DOI:10.13210/j.cnki.jhmu.20240524.002.
基金信息:
国家自然科学基金资助项目（81973844、81573864）~~
2024-05-27
2024-05-27
2024-05-27