摘要
目的探讨久泻灵颗粒对脾肾阳虚型溃疡性结肠炎模型大鼠结肠组织中TLR4、NF-κB p65蛋白基因的表达的影响。方法通过灌服大黄水煎液,肌肉注射氢化可的松并结合TNBS(2,4,6-三硝基苯磺酸)混合乙醇灌肠建立动物模型。将90只大鼠随机分为空白组、模型组、久泻灵颗粒治疗7、14、21 d组及阳性对照(SASP)组,用药治疗后采用免疫组化SP法和RT-q PCR法分别检测大鼠结肠组织中TLR4、NF-κB p65蛋白和基因的表达。结果结肠黏膜形态学评分结果显示,各治疗组评分均有降低,差异有显著性(P<0.01);免疫组化SP法结果显示:TLR4、NF-κB p65在模型组大鼠中表达均显著增强(P<0.01);药物干预后各治疗组大鼠TLR4、NF-κB p65表达均减弱(P<0.05);RT-q PCR法结果显示:与空白组比较,模型组大鼠TLR4、NF-κB p65表达增强(P<0.01);与模型组比较,治疗后各组大鼠TLR4、NF-κB p65表达减弱(P<0.05)。结论久泻灵颗粒可减轻结肠黏膜炎症反应,起到修复黏膜的作用,其原因与降低脾肾阳虚型UC模型大鼠结肠组织中TLR4、NF-κB p65基因的转录和蛋白的表达有关。
Objective To explore the effects of a Chinese medicine,Jiuxieling granules,on the expression of TLR4 and NF-κB p65 m RNA and proteins in ulcerative colitis model rats with spleen and kidney yang deficiency. Methods The rat model of ulcerative colitis with spleen and kidney yang deficiency was established by oral administration of rhubarb decoction,intramuscular injection of hydrocortisone and TNBS（ 2,4,6-trinitrobenzene sulfonic acid） combined with ethanol enema. All the 90 rats were randomly divided into blank control group,UC model of spleen kidney yang deficiency group,and Jiuxieling granules treatment groups for 7 days,14 days and 21 days,respectively,and positive control group（ SASP）. After all the rats were killed and colon samples were collected,the expression of TLR4 and NF-κB p65 m RNA and proteins of colon tissues were detected by RT-q PCR and SP immunohistochemical. Results The results of immunohistochemiscal assay indicated that expressions of TLR4 and NF-κB p65 in the UC model group were significantly enhanced,when compared with that of the blank group（ P〈0. 01）. After treatment,the expressions of TLR4 and NF-κB p65 in all treatment groups were significantly decreased（ P〈0. 01 or P〈0. 05）. The RT-qP CR assay results indicated that the expressions of TLR4 and NF-κB p65 in the UC model group were significantly enhanced when compared with that of the blank group（ P〈0. 01）. After treatment,the expressions of TLR4 and NF-κB p65 in all treatment groups were reduced（ P〈0. 01 or P〈0. 05）. Conclusions Jiuxieling granules can alleviate the mucosal inflammation and facilitate the mucosal repair. The main reasons of which may be associated with reduction of TLR4 and NF-κB p65 gene transcription and protein expression in the ulcerative colitis model rats with spleen and kidney yang deficiency.
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2016年第1期47-52,共6页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金项目(编号:81260520)
