Toll样受体在溃疡性结肠炎患者外周血中的表达及其意义

孙宏飞; 郭莲怡

doi:10.3969/j.issn.2095-5227.2016.06.028

摘要: 目的探讨溃疡性结肠炎(ulcerative colitis，UC)患者外周血单个核细胞Toll样受体(toll-like receptors，TLRs)的表达及其临床意义。方法收集20例溃疡性结肠炎患者治疗前后(急性期与缓解期)的外周血标本，另取20例正常人作为对照。通过RT-PCR方法检测急性期、缓解期溃疡性结肠炎患者和正常人外周血单个核细胞中TLR1、TLR2、TLR3、TLR4、TLR5、TLR9 mRNA水平；用Western blot方法检测急性期、缓解期溃疡性结肠炎患者和正常人外周血单个核细胞中TLR1、TLR2、TLR3、TLR4、TLR5、TLR9的表达；流式细胞术检测急性期、缓解期溃疡性结肠炎患者和正常人外周血单个核细胞中TLR1、TLR2、TLR3、TLR4、TLR5、TLR9的表达，以确定TLRs的来源。结果急性期溃疡性结肠炎患者外周血单个核细胞TLR2、TLR4、TLR5、TLR9 mRNA水平(8.47±1.20，21.05±2.22，9.62±1.10，8.76±1.02)较正常对照组(4.65±0.88，10.79±0.99，4.95±0.78，3.91±0.74)升高(P< 0.05)，TLR1、TLR3 mRNA水平差异无统计学意义(P> 0.05)；缓解期溃疡性结肠炎患者外周血单个核细胞TLR4、TLR5、TLR9 mRNA水平(12.85±0.81，7.49±0.73，6.27±0.62)较正常对照组(10.79±0.99，4.95±0.78，3.91±0.74)升高(P< 0.05)，TLR1、TLR2、TLR3 mRNA水平差异无统计学意义(P> 0.05)。急性期UC患者外周血单个核细胞TLR2、TLR4、TLR5、TLR9 mRNA水平较缓解期高(P< 0.05)。外周血单个核细胞中T细胞、自然杀伤(naturak killer，NK)T细胞、单核细胞和B细胞中均表达TLR1、TLR2、TLR3、TLR4、TLR5、TLR9。结论急性期与缓解期UC患者的TLR2、TLR4、TLR5、TLR9的表达在大部分细胞中呈不同程度的上调，但TLR1、TLR3的表达变化较小。推测TLRs及其介导的天然免疫应答可促进溃疡性结肠炎的发生。

Abstract: Objective To investigate the expression of toll-like receptors (TLRs) in peripheral blood mononuclear cell (PBMC) of patients with ulcerative colitis (UC) and its clinical significance. Methods Blood samples of 20 patients with ulcerative colitis and 20 healthy volunteers control were collected and analyzed.The levels of mRNA in TLR1, TLR2, TLR3, TLR4, TLR5, TLR9 in PBMC of patients with UC in acute and chronic stage and healthy controls were detected by RT-PCR, and the expression of TLR1, TLR2, TLR3, TLR4, TLR5, TLR9 in PBMC of patients with UC in acute and chronic stage and healthy controls were detected by western blot and flow cytometry so as to confirm the origins of TLRs. Results The levels of TLR2, TLR4, TLR5, TLR9 in PBMC of patients with UC in acute stage were significantly higher than healthy control group (8.47±1.20) vs (4.65±0.88), (21.05±2.22) vs (10.79±0.99), (9.62±1.10) vs (4.95±0.78), (8.76±1.02) vs (3.91±0.74), P< 0.05.There was no difference in TLR1, TLR3 between patients with UC in acute stage and healthy control group (4.37±0.61) vs (4.07±0.51), (7.50±0.80) vs (7.430±1.05), P> 0.05.The levels of TLR4, TLR5, TLR9 in PBMC of patients with UC in chronic stage were significantly higher than healthy control group (12.85±0.81) vs (10.79±0.99), (7.49±0.73) vs (4.95±0.78), (6.27±0.62) vs (3.91±0.74), P< 0.05, while there was no difference in TLR1, TLR2, TLR3 between patients with UC in chronic stage and healthy control group (4.30±0.61) vs (4.07±0.51), (4.42±0.86) vs (4.65±0.88), (7.51±0.80) vs (7.43±1.05), P> 0.05.The levels of TLR2, TLR4, TLR5, TLR9 in PBMC of patients with UC in acute stage were higher than patients with UC in chronic stage (P< 0.05).Large proportions of T cells, NKT cells, monocyte and B cells in PBMC expressed TLR1, TLR2, TLR3, TLR4, TLR5, TLR9. Conclusion The expression of TLR2, TLR4, TLR5, TLR9 in patients with UC in acute and chronic stage increases to some extent, which plays an important role in the pathogenesis of UC, while the expression of TLR1, TLR3 changes a little.It suggests that TLRs and the innate immuneresponse mediated by TLRs may promote the occurrence of UC.

Toll样受体在溃疡性结肠炎患者外周血中的表达及其意义

Expression and significance of TLRs in PBMC of patients with ulcerative colitis