修回日期: 2009-06-20
接受日期: 2009-06-29
在线出版日期: 2009-07-28
目的: 评价黄术灌肠液对腹泻型肠易激综合征(Diarrhea- irritable bowel syndrome D-IBS)大鼠血清IL-1β、IL-10表达的影响.
方法: 采用乙酸灌肠加束缚应激法建立D-IBS模型, 将60只健康♀SD大鼠随机分为正常对照组、模型组、HSE灌肠液组、得舒特组, 每组15只, 分别观察治疗后大鼠血清IL-1β、IL-10的表达.
结果: 与正常对照组比较, 模型组IL-1β显著升高、IL-10显著降低(t = 21.9998, t = 27.0556; 均P<0.01), 治疗后HSE灌肠液组和得舒特组IL-1β较模型组显著降低(t = 12.0599, t = 6.1647; 均P<0.01), 而两者IL-10明显升高(t = 17.3802, t = 6.8408; 均P<0.01), 且HSE灌肠液组效果优于得舒特组(t = 9.7410, P<0.01).
结论: HSE灌肠液可能通过降低活化的免疫细胞数目和上调抗炎因子及增强其抗炎活性, 纠正促炎/抗炎因子失衡, 从而达到治疗D-IBS的目的.
引文著录: 胡俊, 胡团敏, 何文钦, 黄永德. 黄术灌肠液对腹泻型肠易激综合征大鼠IL-1β、IL-10表达的影响. 世界华人消化杂志 2009; 17(21): 2188-2191
Revised: June 20, 2009
Accepted: June 29, 2009
Published online: July 28, 2009
AIM: To investigate the effects of Huangshu Enema (HSE) on the levels of IL-1β and IL-10 in the serum of rats with diarrhea-predominant irritable bowel syndrome (D-IBS).
METHODS: A rat model of D-IBS was created by intracolonic instillation of acetic acid and restraint stress. Sixty healthy female Sprague-Dawley rats were randomly divided into normal control group (n = 15), model control group (n = 15), HSE treatment group (n = 15) and dicetel treatment group (n = 15). The changes in serum IL-1β and IL-10 levels in rats were observed.
RESULTS: The level of serum IL-1β was significantly higher in the model control group than the normal control group, while the level of serum IL-10 was significantly lower in the model control group than in the normal control group (t = 21.9998 and 27.0556, respectively; both P < 0.01). The level of serum IL-1β in the two treatment groups significantly decreased when compared to the model control group (t = 12.0599 and 6.1647, respectively; both P < 0.01), while the level of serum IL-10 in the two treatment groups significantly increased when compared to the model control group (t = 17.3802 and 6.8408, respectively; both P < 0.01). HSE was superior to dicetel in decreasing serum IL-1β level and increasing serum IL-10 level (t = 9.7410, P < 0.01).
CONCLUSION: HSE may be able to reduce the number of activated immune cells, increase anti-inflammatory cytokines and enhance the anti-inflammatory activity, and adjust the imbalance between anti-inflammatory and pro-inflammatory cytokines, thereby exerting therapeutic effects on D-IBS.
- Citation: Hu J, Hu TM, He WQ, Huang YD. Effects of Huangshu Enema on the levels of serum IL-1β and IL-10 in rats with diarrhea-predominant irritable bowel syndrome. Shijie Huaren Xiaohua Zazhi 2009; 17(21): 2188-2191
- URL: https://www.wjgnet.com/1009-3079/full/v17/i21/2188.htm
- DOI: https://dx.doi.org/10.11569/wcjd.v17.i21.2188
肠易激综合征(irritable bowel syndrome, IBS)的病因与发病机制目前尚未完全明了, 但越来越多研究表明, 炎症与IBS发病有着密不可分的联系. 研究发现, 5-羟色胺(5-hydroxytryptamine, 5-HT)[1-2]、P物质(substance P, SP)[3]、血管活性肠肽(vasoactive intestinal peptides, VIP)[4]、胆囊收缩素(cholecystokinin, CCK)[5]及白介素(interleukin, IL)[6-7]等作为中间环节, 在IBS的发生和发展中起着重要作用. 促炎因子与抗炎因子失衡可能是IBS发生发展的关键. 本实验主要观察黄术灌肠液对IBS大鼠血清白介素-1β(interleukin-1β, IL-1β)和白介素-10(interleukin-10, IL-10)的影响, 以探讨黄术灌肠液治疗IBS的作用机制.
♀SD大鼠60只, 体质量180±20 g, 购于江西中医学院(动物合格证号: SCXK 2006- 0001); 自组中药方剂黄术灌肠液: 大黄、黄芩、黄连、黄芪、白术各15 g, 加水煎煮至500 mL; 阳性对照药为得舒特, 法国苏威制药, 批号H20040759; 大鼠IL-1β ELISA检测试剂盒(批号: RT110371)、IL-10 ELISA检测试剂盒(批号: RT116876)购于广州齐云生物制剂公司.
1.2.1 分组及造模: 适应性喂养3 d后, 随机分为正常对照组, 模型组, 黄术灌肠液组, 得舒特组, 每组15只. 对模型组、黄术灌肠液组及得舒特组进行造模. 动物模型制作参照文献[8-10]并略加改动. 模型组大鼠经肛门插入石蜡油润滑过的自制导管, 插入深度距肛门8 cm, 缓慢注入40 mL/L的乙酸1 mL, 拔出导管, 将大鼠尾巴抬高并压迫肛门1 min, 然后用0.01 mol/L PBS 1 mL冲洗结肠. 乙酸灌肠后, 用胶带束缚大鼠肩部、前肢及胸部, 限制大鼠用前肢抓搔头面部, 但不限制其活动, 束缚时间为1 h, 之后释放归笼, 充分补液, 自由进食, 造模时间为14 d.
1.2.2 给药: 造模成功后, 黄术灌肠液组以黄术灌肠液3 mL保留灌肠; 得舒特组给予得舒特水溶液0.015 g/(kg•d)灌胃, 模型组及正常对照组以蒸馏水3mL灌胃, 每日1次, 以上均给药14 d.
1.2.3 IL-1β、IL-10表达的检测: 给药结束后第2天, 各组大鼠心脏取血, 3000 r/10 min离心分离血清, ELISA法检测血清IL-1β、IL-10含量. 具体操作严格按照试剂盒说明书.
统计学处理 数据以mean±SD表示, 统计软件处理数据资料, 采用单因素方差分析, 组间两两比较采用t检验.
所有造模大鼠在造模2-3 d后出现腹泻, 呈黄色稀便样和/或肛周体毛被稀便沾染. 5-6 d后体质量略下降, 活动及进食明显减少, 进水增多. 低倍光学显微镜下可见模型组大鼠结肠标本黏膜下层血管轻度扩张, 黏膜完整, 未见溃疡及黏膜增厚, 可见少量炎症细胞浸润, 主要为增多的淋巴细胞及肥大细胞.
黄术灌肠液组及得舒特组用药1 wk后腹泻明显减轻, 进食进水逐渐恢复正常, 用药结束后光学显微镜下见结肠标本淋巴细胞及肥大细胞明显减少.
模型组大鼠较正常组大鼠血清IL-1β表达显著增高(t = 21.9998, P<0.01)且IL-10明显降低(t = 27.0556, P<0.01), 治疗后大鼠血清IL-1β较模型组显著降低(黄术灌肠液组t = 12.0599, P<0.01; 得舒特组t = 6.1647, P<0.01)而IL-10明显升高(黄术灌肠液组t = 17.3802, P<0.01; 得舒特组t = 6.8408, P<0.01), 且黄术灌肠液组效果优于得舒特组(t = 9.7410, P<0.01, 表1).
| 分组 | IL-1β | IL-10 |
| 正常组 | 42.67±4.82 | 177.79±11.99 |
| 模型组 | 104.91±9.84b | 71.66±9.33b |
| 黄术灌肠液组 | 64.38±8.52d | 147.72±14.15d |
| 得舒特组 | 87.42±4.89 | 99.68±12.83 |
长期以来, IBS被认为是一种慢性功能性肠道疾病而不是炎症性疾病, 近年来有学者从IBS患者结肠标本内发现增多的肥大细胞, 从而提出了IBS患者肠壁内可能有炎症成分参与[11]. O'Sullivan et al[12]研究发现绝大多数IBS患者末端回肠和盲肠的肥大细胞数目显著升高. 而Chadwick et al[13]的研究表明: 40%的IBS患者肠黏膜活检组织可见非特异性炎症改变, 而其余患者病理检查虽然正常, 但上皮内淋巴细胞及CD25+/CD3+ T细胞数量显著增加. 随着对PI-IBS的深入研究, 进一步证明了部分IBS患者中存在肠黏膜低度炎症和免疫细胞功能活化.
近年来的研究进一步提示肠道炎症在IBS的发病中起着举足轻重的作用. Törnblom et al[14]研究发现, IBS患者不仅肠黏膜存在轻度炎症和免疫激活, 而且肠神经丛内有淋巴细胞的低度浸润, 并有神经元退化现象. 而胃肠动力障碍及感觉过敏正是以肠神经系统异常为基础[15]. 肠黏膜炎症还可以通过肠道局部的神经内分泌免疫网络的复杂作用引起肠神经系统(enteric nervous system, ENS)-平滑肌功能可逆性的改变, 最终导致肠黏膜化学感受器和机械感受器对各种刺激敏感性增高[16]. 此外, 低度炎症还可以通过改变非炎症部位肠神经反射回路和上皮细胞功能从而引起非炎症部位感觉过敏[17]. 肠道持续低度炎症的原因仍未完全清楚, 促炎因子与抗炎因子表达失衡可能是其重要机制.
IL-1β是重要的促炎因子之一, 也是炎症反应的重要介质, IL-1β可提高胃肠道胆碱能神经对乙酰胆碱及其他炎性介质的敏感性, IL-1β还可抑制肠道Na-K-ATP酶的活性, 进而抑制水、钠吸收而产生腹泻, 最终导致胃肠功能的紊乱. Gwee et al[18]发现急性胃肠炎后发展为IBS患者比急性胃肠炎不进展为IBS者有更高的直肠黏膜IL-1β mRNA表达. 王利华 et al[19]研究也证实了这点. 而IL-10是重要的抗炎因子并与IBS密切相关, Gonsalkorale et al[20]研究发现, IBS患者IL-10基因频率较正常显著降低, 提示IL-10产生减少的个体更容易发生IBS.
本研究中自组中药方剂黄术灌肠液主要成分为大黄、黄芩、白术等, 现代医学研究表明大黄具有抑菌、调节肠管舒缩运动、清除肠道有毒物质活性等作用; 黄芪能清除体内氧自由基、扩张血管、改善微循环及调节机体免疫功能; 白术与黄芪配伍具有协同作用, 可调控肠黏膜细胞炎症修复, 改善肠道功能[21-22]. 我们前期的临床治疗研究亦表明本组方剂配伍合理, 具有明显协同作用, 抗菌消炎, 改善黏膜血循环, 促进炎症修复, 调整肠道功能[23]. 尤其是黄术灌肠液通过灌肠给药高浓度直接作用于病变部位, 扶正祛邪, 促进局部炎症和水肿消退, 缩短病程. 而且灌肠治疗减少了药物在肝脏的首过效应, 有着传统口服给药无法比拟的优势[24]. 本研究结果显示, IBS大鼠促炎因子IL-1β水平升高而抗炎因子IL-10水平降低, 提示促炎/抗炎因子失衡在IBS的发生和发展中占有重要地位. 黄术灌肠液灌肠治疗后显著降低IBS大鼠血清IL-1β及升高IL-10, 且效果明显优于得舒特组, 说明黄术灌肠液可能通过降低活化的免疫细胞数目, 减少IL-1β表达, 上调抗炎因子IL-10及增强其抗炎活性, 从而纠正抗炎/促炎因子失衡, 最终达到对IBS的治疗效果.
肠易激综合征(IBS)是消化科常见疾病, IBS的发病机制复杂, 是多种因素作用的结果. 肠道持续性低度炎症、精神心理因素、内脏高敏感性、胃肠动力异常、感染等因素可能与IBS发病有关.
魏睦新, 教授, 南京医科大学第一附属医院中医内科.
肠道持续性低度炎症与IBS发病之间的联系是近年来研究的热点. 肠道持续性低度炎症的发生机制尚待进一步研究.
传统治疗IBS多以口服药为主, 本研究通过自组中药方剂保留灌肠, 高浓度药物直接作用于病变部位, 疗效显著, 目前国内尚少见类似报道.
本文深入探讨了IBS中促炎因子IL-1β及抗炎因子IL-10表达改变, 为肠道低度炎症在IBS发病中占有重要地位提供了重要依椐, 可能为阐明IBS发病机制及治疗带来新的希望.
本研究设计合理, 内容具有原创性, 学术价值较好.
编辑:李军亮 电编:吴鹏朕
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