溃结灵对溃疡性结肠炎大鼠Th17分化的调节机制研究  

The regulatory mechanism of Kuijieling decoction on Th17 differentiation in ulcerative colitis rats

作　　者：王凤[1] 吴秀美[2] 甘丽萍[1] 龙宇[1] 李燕舞[1] 巫燕莉[1] 刘梓峰[1] 杜群[1] 

机构地区：[1]广州中医药大学脾胃研究所,广州510405 [2]广州中医药大学第一附属医院急诊科,广州510405

出　　处：《中药药理与临床》2016年第2期177-181,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81373798);广州中医药大学中医药防治脾胃病脑病创新研究团队项目(A1-AFD01514A05)

摘　　要：目的:探讨溃结灵对溃疡性结肠炎(UC)大鼠Th17细胞分化相关因子的影响,从而揭示其作用机制。方法:采用雄性SD大鼠制作由三硝基苯磺酸(TNBS)所导致的溃疡性结肠炎模型,溃结灵高、中、低剂量以及柳氮磺胺吡啶(SASP)对溃疡性结肠炎大鼠给予治疗,治疗结束后采集结肠粘膜,采用酶联免疫吸附(ELISA)法检测大鼠结肠粘膜中白细胞介素6(IL-6)、转化生长因子β1(TGF-β1)的含量;采用荧光定量PCR检测大鼠结肠组织IL-6、TGF-β1、孤独核受体t(RORγt)、信号转导子与转录激活子3(STAT3)mRNA的表达;采用Western blot法检测结肠粘膜RORγt、STAT3蛋白表达。结果:溃结灵高剂量(18.3g/kg)和中剂量(9.2g/kg)可明显减少溃疡性结肠炎大鼠结肠的溃疡个数和溃疡面积。大鼠结肠粘膜中,模型组IL-6的含量及mRNA表达明显高于正常组,溃结灵高剂量组(18.3g/kg)和柳氮磺胺吡啶组(0.5g/kg)低于模型组;模型组TGF-β1的含量及mRNA表达明显低于正常组,溃结灵高剂量(18.3g/kg)和柳氮磺胺吡啶组(0.5g/kg)高于模型组。模型组结肠粘膜中RORγt、STAT3蛋白表达及mRNA表达明显高于正常组,溃结灵高(18.3g/kg)、低(4.6g/kg)剂组和柳氮磺胺吡啶组(0.5g/kg)的蛋白表达及mRNA表达低于模型组。结论:溃结灵可能通过调节溃疡性结肠炎大鼠结肠粘膜内与Th17分化相关的IL-6、TGF-β1、RORγt、STAT3的表达,从而抑制炎症反应,治疗溃疡性结肠炎。Objective： To investigate the traditional Chinese medicine compound effect of Kuijieling（ KD） decoction on rat T helper 17（ Th17） cell differentiation related cytokines of ulcerative colitis（ UC）,and to explore its mechanism of action. Methods： The UC models were induced by TNBS using male SD rats. The treatment was given by KD high dose,medium dose and low dose. After treatment the rats were killed to collect their colonic mucosa. Contents of IL-6,TGF-β1 in colonic mucosa of rats were determined by ELISA. The expression levels of IL-6,TGF-β1,the orphan retinoic acid nuclear receptor（ RORγt） and signal transducer and activator of transcription 3（ STAT3） mRNA were detected by real time polymerase chain reaction（ RT-PCR）. The expression of RORγt and STAT3 at protein level by Western blot. Results： The number and area of ulcer in UC rats colon can be obviously reduced by KD high dose group（ 18. 3g / kg） and KD medium dose group（ 9. 2g /kg）. In colonic mucosa of rats,compared to normal controls,the content and mRNA expression of IL-6,in model control group increased（ P〈 0. 05）,whereas the content and mRNA expression of TGF-β1 decreased（ P 〈 0. 05）,and compared to model controls,the content and mRNA expression of IL-6 in KD high dose group（ 18. 3g / kg） and SASP group（ 0. 5g / kg） decreased（ P 〈 0. 05）,whereas the content and mRNA expression of TGF-β1 in KD high dose（ 18. 3g / kg）,and SASP group（ 0. 5g / kg） increased（ P 〈 0. 05）. In colonic tissue,the expression of RORγt and STAT3 mRNA and protein in model control group were significantly higher than those in normal controls（ P 〈 0. 05）,the expression of RORγt and STAT3 mRNA and protein in KD high dose（ 18. 3g / kg）,low dose group（ 4. 6g / kg） and SASP group（ 0. 5g / kg） were lower than those in model group（ P 〈 0. 01 or P 〈 0. 05）. Conlusion： KD cure the UC possibly through regulation the expression of IL-6,TGF-β1,RORγt,STAT3 to inhibition inflammatory

关 键 词：溃结灵 溃疡性结肠炎 TH17 IL-6 TGF-Β1 RORΓT STAT3 

分 类 号：R285.5[医药卫生—中药学]