郁金散对大肠湿热证大鼠回肠、结肠黏膜SIgA和炎性因子的影响

doi:10.11843/j.issn.0366-6964.2019.10.022

摘要/Abstract

摘要： 旨在探讨郁金散对大肠湿热证大鼠回肠、结肠肠黏膜SIgA和炎性因子的影响。体重180~220 g Wistar大鼠40只，随机分为正常对照组、大肠湿热证模型组、自愈组和郁金散治疗组，每组10只。通过饮食设置（高糖高脂饮食）、饥饱失常和灌服白酒并结合高温高湿环境，最后腹腔注射生物因子大肠杆菌，以模拟大肠湿热证发病条件，建立大肠湿热证大鼠模型。郁金散治疗组用郁金散治疗5 d，然后采集回肠和结肠组织制作病理切片，进行组织病理学观察；用ELISA法检测回肠和结肠黏液中免疫球蛋白SIgA含量及组织炎性细胞因子TNF-α、IL-1β、IL-6、IL-10、IL-17和IL-23含量。结果显示，模型大鼠回肠和结肠黏膜上皮细胞变性、坏死、脱落，上皮完整性被破坏，有炎性细胞浸润现象；郁金散治疗组黏膜上皮再生，完整性得到恢复，连续性良好，排列有序，炎性细胞浸润明显减少。模型组大鼠回肠和结肠黏液中的SIgA含量均显著高于正常对照组（P<0.05）；郁金散治疗组显著低于自愈组（P<0.05）。模型组回肠和结肠组织中促炎因子TNF-α、IL-1β、IL-6、IL-17和IL-23含量显著或极显著高于正常对照组（P<0.05或P<0.01），抑炎因子IL-10含量显著或极显著低于正常对照组（P<0.05或P<0.01）；郁金散治疗组回肠组织的上述炎性因子含量与自愈组相比均差异显著（P<0.05）；结肠中的TNF-α、IL-10、IL-23与自愈组差异显著（P<0.05）；IL-1β、IL-6、IL-17与自愈组差异极显著（P<0.01）。结果表明，大肠湿热证模型大鼠回肠和结肠黏膜免疫稳态被打破，表现为SIgA分泌亢进，炎性细胞因子分泌紊乱；郁金散治疗可以促使其显著回调。

Abstract: This study was conducted to explore the effects of Yujin powder (YJP) on SIgA and inflammatory cytokines in ileal and colonic mucosa of Large Intestine Dampness-heat Syndrome (LIDHS) rat. Forty Wistar rats were randomly divided into 4 groups:normal control, LIDHS model, self-healing and YJP treatment group, with 10 rats in each group. The rat model of LIDHS was established through setting such complex factors as high-sugar and high-fat diet, improper diet, high temperature and high humidity environment, drinking and intraperitoneal injection of Escherichia coil to imitate the inducing conditions of LIDHS. After the model being successfully established, the rats of YJP treatment group were treated with YJP for 5 days. And then we collected ileum and colon, made histopathological slices and observed the pathological changes of ileum and colon. Mucous SIgA and tissue TNF-α, IL-1β, IL-6, IL-10, IL-17 and IL-23 in ileum and colon were detected by ELISA method. The results showed that in model rats, the ileal and colonic mucosal epithelium cells showed degeneration, necrosis and exfoliation, the integrality of epithelium was destroyed; and there were inflammatory cells infiltration. In YJP group, mucosal epithelium regenerated, the integrality was recovered, the continuity was good, epithelium cells have a regular arrangement, and inflammatory cells infiltration obviously decreased. The contents of mucous SIgA in ileum and colon in LIDHS model group were significantly higher than those in normal group (P<0.05); those in YJP treatment group were both significantly lower than those in self-healing group (P<0.05). The contents of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, IL-17 and IL-23 in ileum and colon tissue were significantly or extremely significantly higher than those in normal group (P<0.05 or P<0.01). The contents of anti-inflammatory cytokine IL-10 were significantly or extremely significantly lower than those in normal group (P<0.05 or P<0.01). The contents of above inflammatory cytokines in ileum tissue in YJP treatment group were significantly different from those in self-healing group (P<0.05); the contents of TNF-α, IL-10 and IL-23 in colon were significantly different from those in self-healing group (P<0.05); the contents of IL-1β, IL-6 and IL-17 were extremely significantly different from those in self-healing group (P<0.01). It is demonstrated that ileal and colonic mucosal immune homeostasis of LIDHS model rat was destroyed, which manifested as hypersecretion of SIgA and secretion disorder of inflammatory cytokines. The treatment of YJP could back-regulate this phenomenon.

中图分类号:

姚万玲, 文艳巧, 华永丽, 纪鹏, 魏彦明. 郁金散对大肠湿热证大鼠回肠、结肠黏膜SIgA和炎性因子的影响[J]. 畜牧兽医学报, 2019, 50(10): 2147-2155.

YAO Wanling, WEN Yanqiao, HUA Yongli, JI Peng, WEI Yanming. Effects of Yujin Powder on SIgA and Inflammatory Cytokines in Ileal and Colonic Mucosa of Large Intestine Dampness-heat Syndrome Rat[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2147-2155.

参考文献 35

[1]	邓铁涛. 中医诊断学[M]. 2版. 北京:人民卫生出版社, 2008.
	DENG T T. Diagnostics of traditional Chinese medicine[M]. 2nd ed. Beijing:People's Medical Publishing House, 2008. (in Chinese)
[2]	MEBIUS R E. Organogenesis of lymphoid tissues[J]. Nat Rev Immunol, 2003, 3(4):292-303.
[3]	MACPHERSON A J, UHR T. Induction of protective IgA by intestinal dendritic cells carrying commensal bacteria[J]. Science, 2004, 303(5664):1662-1665.
[4]	张志军, 刘懿,王磊,等. TLR4mAb对急性期溃疡性结肠炎小鼠结肠黏膜中促炎因子TNF-α、IFN-γ、IL-1β的影响[J]. 复旦学报:医学版, 2008, 35(2):176-180.
	ZHANG Z J, LIU Y, WANG L, et al. Preventive effects of TLR4 monoclonal antibodies on the gut mucosal proinflammatory cytokines (TNF-α, IFN-γ, IL-1β) expression in mice with DSS-induced acute ulcerative colitis[J]. Fudan University Journal of Medical Sciences, 2008, 35(2):176-180. (in Chinese)
[5]	郑紫丹, 万晓强,刘梁英. 溃疡性结肠炎患者血清IL-23和IL-17的水平变化及意义[J]. 细胞与分子免疫学杂志, 2011, 27(2):203-204.
	ZHENG Z D, WAN X Q, LIU L Y. Serum contents of IL-23 and IL-17 in the patients with ulcerative colitis and the clinical significance[J]. Chinese Journal of Cellular and Molecular Immunology, 2011, 27(2):203-204. (in Chinese)
[6]	YAO W L, YANG C X, WEN Y Q, et al. Treatment effects and mechanisms of Yujin Powder on rat model of large intestine dampness-heat syndrome[J]. J Ethnopharmacol, 2017, 202:265-280.
[7]	王自力, 于同泉,朱晓宇,等. 中药复方对热应激下猪肠道组织IL-2、IL-10和黏液IgA含量影响[J]. 中国兽医杂志, 2007, 43(9):83-85.
	WANG Z L, YU T Q, ZHU X Y, et al. Effects of complex prescriptions of Chinese herbs on the concentration of IL-2, IL-10 and IgA in porcine intestines after heat stress[J]. Chinese Journal of Veterinary Medicine, 2007, 43(9):83-85. (in Chinese)
[8]	黄煌,郑鹏远,罗予,等. 食物过敏对小鼠肠道屏障功能及CD4+CD25+调节性T细胞的影响[J]. 世界华人消化杂志, 2008, 16(17):1932-1937.
	HUANG H, ZHENG P Y, LUO Y, et al. Influence of food allergy on gut barrier function and CD4+CD25+ regulatory T cells in food allergic mice[J]. World Chinese Journal of Digestology, 2008, 16(17):1932-1937. (in Chinese)
[9]	HAPFELMEIER S, LAWSON M A E, SLACK E, et al. Reversible microbial colonization of germ-free mice reveals the dynamics of IgA immune responses[J]. Science, 2010, 328(5986):1705-1709.
[10]	FAGARASAN S, MURAMATSU M, SUZUKI K, et al. Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora[J]. Science, 2002, 298(5597):1424-1427.
[11]	LAMKHIOUED B, GOUNNI A S, GRUART V, et al. Human eosinophils express a receptor for secretory component. Role in secretory IgA-dependent activation[J]. Eur J Immunol, 1995, 25(1):117-125.
[12]	MOTEGI Y, KITA H. Interaction with secretory component stimulates effector functions of human eosinophils but not of neutrophils[J]. J Immunol, 1998, 161(8):4340-4346.
[13]	DE ARAÚJO JÚNIOR R F, DA SILVA REINALDO M P O, DE CASTRO BRITO G A, et al. Olmesartan decreased levels of IL-1β and TNF-α, down-regulated MMP-2, MMP-9, COX-2, RANK/RANKL and up-regulated SOCs-1 in an intestinal mucositis model[J]. PLoS One, 2014, 9(12):e114923.
[14]	MUDTER J, NEURATH M F. IL-6 signaling in inflammatory bowel disease:pathophysiological role and clinical relevance[J]. Inflamm Bowel Dis, 2007, 13(8):1016-1023.
[15]	FUJINO S, ANDOH A, BAMBA S, et al. Increased expression of interleukin 17 in inflammatory bowel disease[J]. Gut, 2003, 52(1):65-70.
[16]	LIU Z J, YADAV P K, XU X R, et al. The increased expression of IL-23 in inflammatory bowel disease promotes intraepithelial and lamina propria lymphocyte inflammatory responses and cytotoxicity[J]. J Leukoc Biol, 2011, 89(4):597-606.
[17]	RADDATZ D, BOCKEMÜHL M, RAMADORI G. Quantitative measurement of cytokine mRNA in inflammatory bowel disease:relation to clinical and endoscopic activity and outcome[J]. Eur J Gastroenterol Hepatol, 2005, 17(5):547-557.
[18]	ZHANG J X, DANG S C, QU J G, et al. Changes of gastric and intestinal blood flow, serum phospholipase A2 and interleukin-1β in rats with acute necrotizing pancreatitis[J]. World J Gastroenterol, 2005, 11(23):3578-3581.
[19]	梁皓天, 宋静荣. TNF-α、IL-6、IL-8水平与溃疡性结肠炎相关性研究[J]. 中国现代药物应用, 2010, 4(21):92.
	LIANG H T, SONG J R. Study on the correlation between the levels of TNF-α, IL-6 and IL-8 and ulcerative colitis[J]. Chinese Journal of Modern Drug Application, 2010, 4(21):92. (in Chinese)
[20]	ZHANG F, YAO S M, ZHANG M X, et al. Roles of circulating soluble interleukin (IL)-6 receptor and IL-6 receptor expression on CD4+ T cells in patients with chronic hepatitis B[J]. Int J Infect Dis, 2011, 15(4):e267-e271.
[21]	PAUL A T, GOHIL V M, BHUTANI K K. Modulating TNF-α signaling with natural products[J]. Drug Discov Today, 2006, 11(15-16):725-732.
[22]	KOLLS J K, LINDÉN A. Interleukin-17 family members and inflammation[J]. Immunity, 2004, 21(4):467-476.
[23]	肖南平, 欧阳钦. IL-23/IL-17轴与炎症性肠病[J]. 国际消化病杂志, 2008, 28(3):234-236.
	XIAO N P, OUYANG Q. The interleukin-23/17 axis and inflammatory bowel disease[J]. International Journal of Digestive Diseases, 2008, 28(3):234-236. (in Chinese)
[24]	CASTRO-SANTOS P, SUAREZ A, MOZO L, et al. Association of IL-10 and TNFα genotypes with ANCA appearance in ulcerative colitis[J]. Clin Immunol, 2007, 122(1):108-114.
[25]	KANT V, GOPAL A, PATHAK N N, et al. Antioxidant and anti-inflammatory potential of curcumin accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats[J]. Int Immunopharmacol, 2014, 20(2):322-330.
[26]	DOMENICO C, LINSENBARDT H, HANEY R, et al. (544) Evaluating the anti-hyperalgesic and anti-inflammatory potential of curcumin in a human model of experimental pain[J]. J Pain, 2015, 16(S4):S112.
[27]	NAIR V, SINGH S, GUPTA Y K. Anti-arthritic and disease modifying activity of Terminalia chebula Retz. in experimental models[J]. J Pharm Pharmacol, 2010, 62(12):1801-1806.
[28]	LEE W, KU S K, BAE J S. Anti-inflammatory effects of Baicalin, Baicalein, and Wogonin in vitro and in vivo[J]. Inflammation, 2015, 38(1):110-125.
[29]	CHOI R J, NGOC T M, BAE K H, et al. Anti-inflammatory properties of anthraquinones and their relationship with the regulation of P-glycoprotein function and expression[J]. Eur J Pharm Sci, 2013, 48(1-2):272-281.
[30]	HU B Y, ZHANG H, MENG X L, et al. Aloe-emodin from rhubarb (Rheum rhabarbarum) inhibits lipopolysaccharide-induced inflammatory responses in RAW264.7 macrophages[J]. J Ethnopharmacol, 2014, 153(3):846-853.
[31]	WU J S, ZHANG H, HU B Y, et al. Coptisine from Coptis chinensis inhibits production of inflammatory mediators in lipopolysaccharide-stimulated RAW 264.7 murine macrophage cells[J]. Eur J Pharmacol, 2016, 780:106-114.
[32]	ZHANG W, DAI S M. Mechanisms involved in the therapeutic effects of Paeonia lactiflora Pallas in rheumatoid arthritis[J]. Int Immunopharmacol, 2012, 14(1):27-31.
[33]	WANG Y N, ZHANG H, DU G H, et al. Total glucosides of paeony (TGP) inhibits the production of inflammatory cytokines in oral lichen planus by suppressing the NF-κB signaling pathway[J]. Int Immunopharmacol, 2016, 36:67-72.
[34]	HAN Y, XU L, WANG Q, et al. Chemical composition, antioxidant and antimicrobial activity of the essential oil of phellodendron amurense (Rupr.) from China[J]. Asian J Chem, 2015, 27(3):841-844.
[35]	CHOI Y Y, KIM M H, HAN J M, et al. The anti-inflammatory potential of Cortex Phellodendron in vivo and in vitro:down-regulation of NO and iNOS through suppression of NF-κB and MAPK activation[J]. Int Immunopharmacol, 2014, 19(2):214-220.