清肠化湿方对溃疡性结肠炎NF-κB/Tolls通路的影响及其机制  

Effect of Qingchang Huashi Recipe on NF-κB/Tolls Pathway in Ulcerative Colitis Patients and Mechanism Study

作　　者：沈洪 刘智群[2] 朱荃[3] 朱磊 翟金海[4] 

机构地区：[1]南京中医药大学附属医院,江苏省中医院消化科,南京210029 [2]南京市中医院消化科,南京210001 [3]澳门科技大学中医药学院,中国澳门 [4]南京中医药大学第一临床医学院,南京210046

出　　处：《中国中西医结合杂志》2013年第9期1216-1220,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81072778);江苏省自然科学基金资助项目(No.BK2011078);江苏高校优势学科建设工程资助项目(No.B222);江苏省中医药局课题资助项目(No.JD11002,JD11011)

摘　　要：目的观察清肠化湿方对人结肠癌上皮细胞株(HT-29细胞)核因子-κB(nuclear factorκB,NF-κB)、Toll样受体(Toll-like receptors,TLRs)蛋白的活化、表达以及白介素-8(interleukin-8,IL-8)含量的影响,探讨其治疗溃疡性结肠炎(ulcerative colitis,UC)的可能机制。方法予肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、脂多糖(lipopolysaccharides,LPS)诱导HT-29细胞炎症模型,实验分为空白对照组,模型对照组,柳氮磺胺吡啶(sulfasalazine,SASP)组,清肠化湿方低、中、高剂量组。采用MTT法检测实验药物对细胞生长的影响,用Transwell观察巨噬细胞趋化情况,免疫细胞荧光法检测NF-κB、TLR4蛋白,ELISA法检测IL-8含量。结果各实验药物未见抑制细胞生长。清肠化湿方各剂量组在抑制巨噬细胞趋化、减少NF-κB活化,降低TLR4表达,减轻IL-8分泌方面,与模型对照组比较差异均有统计学意义(P<0.05),清肠化湿方高剂量组对巨噬细胞趋化的抑制作用与空白对照组比较差异无统计学意义(P>0.05),而抑制NF-κB活化入核的作用高于SASP组(P<0.05)。结论清肠化湿方可明显减轻HT-29细胞炎症反应,抑制巨噬细胞趋化,减少NF-κB活化入核、降低TLR4的表达,减轻IL-8的分泌,这可能是其治疗UC的作用机制之一。Objective To observe the effect of Qingchang Huashi Recipe （QHR） on the activation and expressions of nuclear factor KB （NF-KB）, Toll-like receptors （TLRs）, and contents of interleukin-8 （IL-8）, thus exploring its possible mechanisms for treating ulcerative colitis （UC）. Methods The HT-29 cells were induced to inflammation model by tumor necrosis factor-α （TNF-α） and lipopolysaccharides （LPS）. HT-29 cells were divided into 6 groups, i.e., the vehicle control group, the model control group, the sulfasalazine （SASP） group, the high dose QHR group, the middle dose QHR group, the low dose QHR group. Effects on the cell growth were detected by MTT. The chemoattractant of macrophages was ob- served using Transwell. The expressions of NF-KB and TLR4 protein were detected using immune cell fluo- rescence method. The content of IL-8 was detected by ELISA. Results ted in each group. Statistical difference existed in each dose QHR group The growth of cells were not inhibi- in inhibiting the chemoattractant of macrophages, reducing activation of NF-KB, lowing expressions of TLR4 protein, and decreasing the se- cretion of IL-8, when compared with the model control group （P 〈0.05）. No statistical difference existed in inhibiting the chemoattractant of macrophages between the high dose QHR group and the vehicle control group （P 〉0.05）. But its inhibition on NF-KB activation was higher in the high dose QHR group than in the SASP group （P 〈0.05）. Conclusion QHR could obviously attenuate the inflammatory reaction of HT-29 cells, inhibit the chemoattractant of macrophages, reduce the activation of NF-KB, lower expressions of TLR-4, and attenuate the secretion of IL-8, which might be one of its mechanisms for treating UC.

关 键 词：清肠化湿方 溃疡性结肠炎 核因子-ΚB TOLL样受体-4 白介素-8 

分 类 号：R285.5[医药卫生—中药学]