紫锥菊提取物对大鼠湿热泄泻的疗效及其机制

doi:10.11843/j.issn.0366-6964.2023.01.035

摘要/Abstract

摘要： 本研究通过建立大鼠湿热泄泻模型，探讨紫锥菊提取物(EPE)对湿热泄泻大鼠的疗效，为紫锥菊提取物防治湿热泄泻提供科学依据。采用“内外湿热+大肠杆菌”建立大鼠湿热泄泻，分别给予1、3g·kg^-1剂量EPE治疗，采用临床症状评分评价疾病模型，测定各组大鼠血常规、血生化、血清炎性因子、脏器指数与结肠相关基因表达水平，并对大鼠器官进行病理组织学观察。结果表明，造模后大鼠被毛蓬松、精神萎靡，扎堆，懒动，眼周出现分泌物，大便溏泄，色黄恶臭，肛周污秽。给药6d后，高、低剂量EPE组与阳性药物组大鼠精神、饮食及临床表现均恢复正常。与空白组相比，模型组大鼠的心、脾、肺指数显著上升(P＜0.05)，血液中红细胞数(RBC)、血红蛋白浓度(HGB)、红细胞压积(HCT)显著降低(P＜0.05)；血清中尿素、丙氨酸氨基转移酶(ALT)水平显著降低(P＜0.05)，IL-6、IL-17、IL-23水平显著上升(P＜0.05)，TGF-β、IL-2、IL-10水平显著下降(P＜0.05)；结肠TGF-β1与Foxp3 mRNA表达量显著下降(P＜0.05)，RORγt mRNA表达显著上升(P＜0.05)；心出现大面积出血瘀血，肝水肿，脾水肿及陈旧性出血，肾出现炎性细胞增生与蛋白尿的沉积。与模型组相比，高、低剂量EPE组的心指数、脾指数、肺指数显著降低(P＜0.05)，血液HCT显著升高(P＜0.05)，血清尿素、总胆固醇(TC)、总蛋白(TP)、白蛋白(ALB)水平显著升高(P＜0.05)，血清IL-17、IL-23水平显著下降(P＜0.05)，TGF-β水平显著上升(P＜0.05)；结肠TGF-β1与Foxp3 mRNA表达显著上升(P＜0.05)，RORγt mRNA表达显著下降(P＜0.05)。高、低剂量EPE组的脏器细胞损伤有所恢复。紫锥菊提取物能通过降低血脂、调节血清炎性因子水平及改善脏器功能以缓解大鼠湿热泄泻。

关键词: 紫锥菊提取物, 湿热泄泻, 大鼠

Abstract: This study established a rat model of damp-heat diarrhea (DHD) to explore the efficacy of Echinacea purpurea extract (EPE) on DHD, so as to provide scientific basis for EPE in the prevention and treatment of DHD. Rats with DHD were established by “internal and external Dampness-heat+Escherichia coli”, and were treated with 1 and 3 g·kg^-1 doses of EPE, respectively. The DHD model was evaluated by clinical symptom score, the blood routine test, blood biochemistry, serum inflammatory factors, organ index and colon related gene expression level of rats in each group were measured, and the histopathology of rat organs was observed. The results showed that the DHD rats were fluffy, depressed, piled up, lazy, secretions appeared in the eyes, loose stool with yellow and smelly, and perianal dirt was visible. After 6 days of administration, the spirit, diet and appearance of rats returned to normal in the high and low dose EPE groups and the positive drug group. Compared with the control group, the indexes of heart, spleen and lung in the model group increased significantly (P<0.05), and the numbers of red blood cell (RBC), hemoglobin (HGB) and hematocrit (HCT) in blood decreased significantly (P<0.05); the levels of urea and alanine aminotransferase (ALT) in serum decreased significantly (P<0.05), the levels of IL-6, IL-17 and IL-23 increased significantly (P<0.05). The levels of TGF-β, IL-2 and IL-10 decreased significantly (P<0.05); the expression of colon TGF-β1 and Foxp3 mRNA decreased significantly (P<0.05), the expression of RORγt mRNA increased significantly (P<0.05); Massive bleeding and stasis in the heart, edema in the liver, edema and old bleeding in the spleen, inflammatory cell proliferation and deposition of proteinuria in the kidneys. Compared with the model group, the heart, spleen and lung of the high and low dose EPE groups were significantly lower (P<0.05), the level of HCT in blood was significantly higher (P<0.05), the levels of urea, total cholesterol (TC), total protein (TP) and albumin (ALB) in serum were significantly higher (P<0.05), the levels of IL-17 and IL-23 in serum were significantly lower (P<0.05), the level of TGF-β increased significantly (P<0.05); The expression of colon TGF-β1 and Foxp3 mRNA increased significantly (P<0.05), the expression of RORγt mRNA decreased significantly (P<0.05). The organ cell damage in high and low dose EPE groups recovered. The data may be extrapolated to suggest that Echinacea purpurea extract can alleviate DHD in rats by improving organ function, reducing blood lipid and regulating the level of serum inflammatory factors.

Key words: Echinacea purpurea extract, damp-heat diarrhea, rats

中图分类号:

S853.74

池幸子, 杨诗靖, 杨泊文, 孙晗, 姚丽丽, 顾达星, 郭世宁, 石达友, 武力, 刘翠. 紫锥菊提取物对大鼠湿热泄泻的疗效及其机制[J]. 畜牧兽医学报, 2023, 54(1): 380-391.

CHI Xingzi, YANG Shijing, YANG Bowen, SUN Han, YAO Lili, GU Daxing, GUO Shining, SHI Dayou, WU Li, LIU Cui. Effect and Mechanism of Echinacea purpurea Extract in Rats with Dampness-heat Diarrhea[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 380-391.

参考文献 30

[1]	吴仕九, 杨运高, 杨钦河, 等. 清热利湿法对湿热证动物模型的影响[J]. 中国中西医结合脾胃杂志, 1999, 7(4):200-202.WU S J, YANG Y G, YANG Q H, et al. Effect of heatness-clearing and wetness-removing therapy on animal model of wetness-heatness syndrome[J]. Chinese Journal of Integrated Traditional and Western Medicine on Gastro-Spleen, 1999, 7(4):200-202. (in Chinese)
[2]	SHEN J, CHEN J J, ZHANG B M, et al. Baicalin is curative against rotavirus damp heat diarrhea by tuning colonic mucosal barrier and lung immune function[J]. Dig Dis Sci, 2020, 65(8):2234-2245.
[3]	廖一羲, 安娣, 石正伟. 中医湿热证动物模型研究进展与问题[J]. 实用中医内科杂志, 2020, 34(12):94-97.LIAO Y X, AN D, SHI Z W. Research progress and problems of animal models of damp-heat syndrome in traditional Chinese medicine[J]. Journal of Practical Traditional Chinese Internal Medicine, 2020, 34(12):94-97. (in Chinese)
[4]	王涛. 两种不同方法建立大鼠湿热泄泻模型的比较研究[D]. 兰州:甘肃农业大学, 2016.WANG T. Competitive study of dampHeat diarrhea rat model estaished with two different methods[D]. Lanzhou:Gansu Agricultural University, 2016. (in Chinese)
[5]	杨巧芳, 孟庆刚, 常辉东, 等. 湿热证动物模型的研究概况[J]. 中华中医药学刊, 2007, 25(10):2057-2059.YANG Q F, MENG Q G, CHANG H D, et al. Overviews on the animal model of damp-heat syndrome[J]. Chinese Archives of Traditional Chinese Medicine, 2007, 25(10):2057-2059. (in Chinese)
[6]	张烁, 张建军, 赵艺萌, 等. 外来植物药紫锥菊文献研究及其"中药化"理论探讨[J]. 中国中药杂志, 2020, 45(5):978-983.ZHANG S, ZHANG J J, ZHAO Y M, et al. Study on literature of Echinacea purpurea and discussion on its herbalization[J]. China Journal of Chinese Materia Medica, 2020, 45(5):978-983. (in Chinese)
[7]	BURGER R A, TORRES A R, WARREN R P, et al. Echinacea-induced cytokine production by human macrophages[J]. Int J Immunopharmacol, 1997, 19(7):371-379.
[8]	LINDE K, BARRETT B, WÖLKART K, et al. Echinacea for preventing and treating the common cold[J]. Cochrane Database Syst Rev, 2006, 1(1):CD000530.
[9]	BARRETT B. Medicinal properties of Echinacea:a critical review[J]. Phytomedicine, 2003, 10(1):66-86.
[10]	SPARREBOOM A, COX M C, ACHARYA M R, et al. Herbal remedies in the United States:potential adverse interactions with anticancer agents[J]. J Clin Oncol, 2004, 22(12):2489-2503.
[11]	PERRY N B, BURGESS E J, GLENNIE V L. Echinacea standardization:analytical methods for phenolic compounds and typical levels in medicinal species[J]. J Agric Food Chem, 2001, 49(4):1702-1706.
[12]	HAGMVLLER W, STOCKHAMMER S, VIELHABER B, et al. Effects of phytogenous industrial waste originating from echinacea, larch and carrot in the prophylaxis of E. coli-diarrhoea in weaner piglets[J]. Wien Tierärztl Monatsschr, 2011, 98(7-8):166-174.
[13]	慕澜, 魏连波, 罗炳德, 等. 大肠湿热证模型大鼠血清部分白细胞介素含量的变化[J]. 中国中西医结合消化杂志, 2003, 11(5):262-263.MU L, WEI L B, LUO B D, et al. Changes of IL-1, IL-2, IL-6 in rats with experimental large intestine damp-heat syndrome model[J]. Chinese Journal of Integrated Traditional and Western Medicine on Digestion, 2003, 11(5):262-263. (in Chinese)
[14]	姚万玲. 大肠湿热证实质及郁金散对其治疗作用机理研究[D]. 兰州:甘肃农业大学, 2017.YAO W L. Essence of large intestine dampness-heat syndrome and treatment mechanisms of Yujin powder on it[D]. Lanzhou:Gansu Agricultural University, 2017. (in Chinese)
[15]	GRVTZNER N. Medicinal plants-an underestimated option to treat gastrointestinal diseases in pigs?[J]. Vet Rec, 2019, 184(10):314-315.
[16]	陈爽白, 常淑枫, 肖照岑, 等. 湿热证大鼠模型的复制及三仁汤对其影响的实验观察[J]. 天津中医, 2002, 19(2):38-40.CHEN S B, CHANG S F, XIAO Z C, et al. Reproduce rat model of dampness-heat syndrome and experimental observation on the effect of San Ren Tang (decoction of three kinds of kernels)[J]. Tianjin Journal of Traditional Chinese Medicine, 2002, 19(2):38-40. (in Chinese)
[17]	郑华, 朱买勋, 张邑帆, 等. 连梅止痢口服液对仔猪湿热泄痢的防治效果研究[J]. 中兽医医药杂志, 2015, 34(4):27-30.ZHENG H, ZHU M X, ZHANG Y F, et al. Study on the preventive effect of Lianmeizhili Oral Solution on hot and humid diarrhea in piglets[J]. Journal of Traditional Chinese Veterinary Medicine, 2015, 34(4):27-30. (in Chinese)
[18]	李学, 魏连波, 罗炳德, 等. 中医大肠湿热证模型SD大鼠肠道的病理变化[J]. 河北中医药学报, 2004, 19(1):4-5.LI X, WEI L B, LUO B D, et al. Enteric pathologic changes of SD rat models with large intestinal damp-heat[J]. Journal of Hebei Traditional Chinese Medicine and Pharmacology, 2004, 19(1):4-5. (in Chinese)
[19]	HARTMAN T L, WATSON K M, YANG L, et al. Broad anti-infective activity of viracea, an Echinacea-derived product[J]. Antiviral Res, 2008, 78(2):A40.
[20]	HILL L L, FOOTE J C, ERICKSON B D, et al. Echinacea purpurea supplementation stimulates select groups of human gastrointestinal tract microbiota[J]. J Clin Pharm Ther, 2006, 31(6):599-604.
[21]	DALBY-BROWN L, BARSETT H, LANDBO A K R, et al. synergistic antioxidative effects of alkamides, caffeic acid derivatives, and polysaccharide fractions from Echinacea purpurea on in vitro oxidation of human low-density lipoproteins[J]. J Agric Food Chem, 2005, 53(24):9413-9423.
[22]	YAN J B, LUO M M, CHEN Z Y, et al. The Function and role of the Th17/Treg cell balance in inflammatory bowel disease[J]. J Immunol Res, 2020, 2020:8813558.
[23]	CHEN Y F, ZHENG J J, QU C, et al. Inonotus obliquus polysaccharide ameliorates dextran sulphate sodium induced colitis involving modulation of Th1/Th2 and Th17/Treg balance[J]. Artif Cells Nanomed Biotechnol, 2019, 47(1):757-766.
[24]	YAO J, GAO R Y, LUO M H, et al. Close homolog of L1-deficient ameliorates inflammatory bowel disease by regulating the balance of Th17/Treg[J]. Gene, 2020, 757:144931.
[25]	刘小玉, 李玉娟, 姜厚望, 等. 叶天士与薛生白治疗湿热下利的用药特色分析[J]. 中国中药杂志, 2018, 43(8):1720-1725.LIU X Y, LI Y J, JIANG H W, et al. Comparison between Ye Tianshi and Xue Shengbai in treatment of diarrfea with damp-heat[J]. China Journal of Chinese Materia Medica, 2018, 43(8):1720-1725. (in Chinese)
[26]	VELIKOVA T V, MITEVA L, STANILOV N, et al. Interleukin-6 compared to the other Th17/Treg related cytokines in inflammatory bowel disease and colorectal cancer[J]. World J Gastroenterol, 2020, 26(16):1912-1925.
[27]	YU R, ZUO F L, MA H Q, et al. Exopolysaccharide-producing Bifidobacterium adolescentis strains with similar adhesion property induce differential regulation of inflammatory immune response in Treg/Th17 axis of DSS-colitis mice[J]. Nutrients, 2019, 11(4):782.
[28]	陈阳, 崔波, 范燕豪, 等. 葛根芩连汤中各成分在正常与大肠湿热证模型大鼠的肠吸收差异研究[J]. 中国中药杂志, 2020, 45(1):169-178.CHEN Y, CUI B, FAN Y H, et al. Differences in intestinal absorption of Gegen Qinlian Decoction between normal rats and rats with large intestinal damp-heat syndrome[J]. China Journal of Chinese Materia Medica, 2020, 45(1):169-178. (in Chinese)
[29]	DYBSKA E, ADAMS A T, DUCLAUX-LORAS R, et al. Waiting in the wings:RUNX3 reveals hidden depths of immune regulation with potential implications for inflammatory bowel disease[J]. Scand J Immunol, 2021, 93(5):e13025.
[30]	HOU R R, XU T L, LI Q, et al. Polysaccharide from Echinacea purpurea reduce the oxidant stress in vitro and in vivo[J]. Int J Biol Macromol, 2020, 149:41-50.