3首止泻名方治疗溃疡性结肠炎大鼠的相关机制对比分析

目的:对比研究黄芩汤、四神丸、痛泻要方3首止泻名方对大鼠溃疡性结肠炎(UC)炎症反应、水运调节及脑肠轴的影响。方法:42只SPF级雄性SD大鼠,以2,4,6-三硝基苯磺酸(TNBS)复合造模法制备UC大鼠模型,按体质量随机分为正常组、模型组、阳性药柳氮磺胺吡啶组(SASP,0.5 g·kg-1),黄芩汤组(Huangqintang,HQT,20 g·kg-1),四神丸组(Sishen Wan,SSW,26 g·kg-1)和痛泻要方组(Tongxie Yaofang,TXYF,22 g·kg-1)。连续给药5 d后,大鼠眼眶取血并处死取结肠,苏木素-伊红(HE)染色检测大鼠结肠组织病变损伤程度,酶联免疫吸附测定(ELISA)结合免疫组化(IHC)测定大鼠血清及结肠组织内血管活性肠肽(VIP),5-羟色胺(5-HT),P物质(SP)分布,蛋白免疫印迹法(Western blot)检测大鼠结肠组织内水通道蛋白3(AQP3),水通道蛋白4(AQP4)蛋白水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠结肠组织内细胞外调节蛋白激酶1(Erk1),p38丝裂原活化蛋白激酶(p38 MAPK)mRNA表达。结果:与正常组比较,模型组5-HT,VIP含量均显著下降(P0.01),SP含量降低,但未见显著性统计学差异;与模型组比较,SASP组和TXYF组5-HT含量明显升高(P0.05),SASP组,HQT组,TXYF组VIP,SP含量均明显升高(P0.05)。与正常组比较,模型组AQP3含量显著升高(P0.01),AQP4含量显著降低(P0.01);与模型组比较,HQT组AQP3含量显著降低(P0.01),SASP组,HQT组AQP4含量明显升高(P0.05);与模型组比较,各给药组Erk1,p38 MAPK mRNA表达均显著降低(P0.01),HQT组降低趋势最为显著。结论:3首名方对大鼠溃疡性结肠炎均有一定治疗作用,可有效改善UC大鼠的体征及精神状况,但在改善泄泻状态下脑肠轴紊乱方面,TXYF治疗作用优于HQT和SSW,在抑制炎症反应及调节水运平衡方面,HQT治疗作用优于TXYF和SSW。

Comparison of Related Mechanism in Rats with Ulcerative Colitis After Treatment of Three Regiments

Objective This study was designed to compare inflammatory response, water carriage and gut brain axis in rats with ulcerative colitis (UC) after treatment of three regiments, Huangqintang (HQT), Sishenwan (SSW), and Tongxie Yaofang(TXYF).Method After approved by Institute of Chinese Materia Medica Ethics Committees in China Academy of Chinese Medical Sciences, UC in rats was induced by using a compound method (trinitrobenzenesulfonic acid plus ethanol). Rats were randomly divided into control, disease, positive control salazosulfapyridine (SASP, 0.5 g·kg^-1), HQT (20 g·kg^-1), SSW(26 g·kg^-1), and TXYF group(22 g·kg^-1). After 5 days of treatment, colonic tissues and the blood were taken for various assays. Damage of colonic tissues was detected by hematoxylin-eosin staining (HE). The distribution of Vasoactine intrestinal (VIP), 5-hydroxytrytamine (5-HT), P-substance (SP) in the blood and serum were detected by enzymelinked immunosorbent assay (ELISA) and immunohistochemistry (IHC), the levels of aquaporin3 (AQP3) and Aquaporin4 (AQP4) in the serum were detected by Western blot, the mRNA expression of Extracellular regulated protein kinases 1 (Erk1) and p38 mitogen-activated protein kinase (p38 MAPK) in the serum were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR).Result The brain-gut peptide results showed that compared with the normal group, the content of 5-HT and VIP in model group were significantly decreased (P<0.01), the content of SP were decreased, but there was no significant statistical difference, compared with the disease group, the content of 5-HT in SASP and TXYF group were clearly increased (P<0.05), the increment of VIP and SP in SASP, HQT, TXYF group were significant (P<0.05). Compared with the normal group, the content of AQP3 in model group were significantly increased(P<0.01), the content of AQP4 were clearly decreased(P<0.01), compared with the disease group, the content of AQP4 in SASP and HQT group were clearly increased (P<0.05), whereas the levels of AQP3 in HQT group were most significant reduced (P<0.01). Compared with the disease group, the expression of Erk1 and p38 were clearly reduced (P<0.01), with the most significant reduce being the expression in HQT group.Conclusion Three regiments all have therapeutic effects on UC, manifested by improvements of the signs and mental status of UC rats. However, in terms of gut-brain axis disturbance improvement, the therapeutic effect of TXYF was superior than HQT and SSW, whereas in terms of inflammatory response suppression and water carriage accomodation, the therapeutic effect of HQT was superior than SSW and TXYF.