Objective The herbal prescription Jiechangling is derived from experience of Professor PAN Jinyao,a famous doctor of Traditional Chinese Medicine in Guangdong province , for the treatment of syndrome of liver depression and qi stagnation,spleen deficiency syndrome of irritable bowel syndrome (IBS). In this study we investigated the effects of Jiechangling on the intestinal mucosal immunization barrier in a rat post-inflammation IBS (PI-IBS) model. Methods A rat model of PI-IBS was established by using a multi-stimulation paradigm including mother-infant separation, bondage and intrarectal administration of TNBS. Four weeks after TNBS administration, the rats were treated with Jiechangling(component: Tangshen, Poria, Hedyotis, Polygonum sinense L, Golden Thread, Debark Peony Root, Jasmine, Orange Fruit, Glutinosae Rice Root, Liquorice Root) (2.85, 5.71 and 11.42 g/kg·d,) for 14 days. Intestinal sensitivity was assessed by the abdominal withdrawal reflex (AWR) scores and fecal water content. Open field test and two-bottle sucrose intake test were used to evaluate the behavioral changes. CD4+ and CD8+ cells were counted and IL-1β and IL-4 levels in intestinal mucosa were measured.Transmission electron microscopy was used to evaluate ultra-structural changes of the intestinal mucosal barrier. Results PI-IBS model rats showed AWR reactivity and fecal water content increased significantly,and locomotor activity and sucrose intake decreased. Jiechangling treatment not only reduced AWR reactivity and fecal water content, but also suppressed the anxiety and depressive behaviors. Ultra-structural study revealed that the gut mucosal barrier function in PI-IBS model rats was severely damaged, whereas Jiechangling treatment relieved intestinal mucosal inflammation and repaired the gut mucosal barrier. Furthermore, CD4+/CD8+ cell ratio in lamina propria and submucosa reduced significantly, and IL-1β and reduced IL-4 expression in intestinal mucosa increased in PI-IBS model rats, whereas Jiechangling treatment reversed PI-IBS-induced changes in CD4+/CD8+ cell ratio and expression of IL-1β and IL-4. Conclusion Jiechangling treatment protects the intestinal mucosa from PI-IBS with anti-inflammatory, immunomodulatory and anti-anxiety effects.%目的 本研究探讨了结肠灵对感染后肠易激综合征(PI-IBS)大鼠模型的结肠黏膜免疫屏障的功效作用.方法 采用复合刺激的造模方法建立肝郁脾虚的PI-IBS大鼠模型,包括母子分离、束缚和三硝基苯磺酸(TNBS)直肠刺激.复合刺激4周后,用结肠灵方治疗,取生药量为2.85、5.71及11.42 g/(kg·d)灌胃14d.用腹壁回缩反射(AWR)评分和粪便含水量评估大鼠的肠内敏感性.旷场实验和双瓶蔗糖汲取实验用来评价大鼠行为改变.测量肠粘膜中的CD4+和CD8+细胞计数以及IL-1β和IL-4水平.透射电镜用来观察肠粘膜的免疫屏障内超微结构改变.结果 造模后模型组大鼠的AWR评分(0.29±0.09)mL和粪便含水量(52.31％±20.12％)明显高于正常组(0.67±0.11)mL和(40.33％±8.34％),模型组大鼠的自发活动(50.24±10.42)和蔗糖汲取(48.98±17.89)明显低予正常组(95.34±14.32)和(75.45±15.98),差异显著有统计学意义(P＜0.05);结肠灵治疗后,高剂量组大鼠的AWR评分(0.59±0.10)mL和粪便含水量(38.34％±6.98％)明显低于模型组;高剂量组大鼠的自发活动(89.63±16.98)和蔗糖汲取(73.42±14.32)明显高于模型组,差异显著有统计学意义(P＜0.05);透射电镜观察到PI-IBS模型大鼠的肠粘膜屏障功能损伤严重,而结肠灵可以缓解肠粘膜炎症并修复肠粘膜屏障.此外,显微镜下观察到肠粘膜的IL-1β水平升高和IL-4水平降低,PI-IBS模型大鼠固有层和粘膜下层的CD4+/CD8+比值明显降低,结肠灵治疗恢复了上述CD4+/CD8+细胞比值、IL-1β和IL-4水平的变化.结论 结肠灵能通过缓解炎症、调节免疫以及舒缓焦虑等作用机制来保护肠粘膜,从而有利于治疗PI-IBS.
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