基于TLR4/NF-κB通路研究白术-白芍药对对IBS-D模型大鼠的治疗作用

Investigating the Therapeutic Effect of Atractylodis Macrocephalae-Paeonia Lactiflora Pairs on IBS-D Model Rats Based on the TLR4-NF-κB Pathway

目的研究白术-白芍药对对腹泻型肠易激综合征(Diarrhea-predominant irritable bowel syndrome,IBS-D)模型大鼠TLR4/NF-κB信号通路相关靶标的影响,旨在阐述其治疗低度炎性反应的潜在机制。方法以番泻叶灌胃+慢性不可预知刺激建立IBS-D模型大鼠,并随机分为模型组、药对组、白芍组、白术组、西药组,每组6只。另取6只不造模大鼠,设为空白组。各药物组大鼠灌胃相应药液,连续治疗14 d。通过体重、大便含水量、AWR评分评价白术-白芍药对对模型大鼠症状的干预效果;各组大鼠腹主动脉取血,剖取回肠末端组织,通过qRT-PCR法、HE染色、Elisa、Westernblot法检测白术-白芍药对对IBS-D大鼠炎症以及TLR4/NF-κB信号通路相关靶标的调控。结果与模型组对比,药对能改善体重、降低大便含水量(P<0.01)、糖水偏好率(P<0.01),在40 mmHg、60 mmHg压力下降低AWR评分(P<0.01),改善回肠末端组织炎症病理状态,下调TLR4、NF-κB、IL-1βmRNA表达(P<0.01),下调血清TLR4、NF-κB、MyD88表达,上调血清IL-10表达,下调MyD88、p-NF-κB p65、IL-6蛋白表达(P<0.01)。结论白术-白芍药对改善IBS-D大鼠腹泻症状和内脏高敏感性,可能与抑制TLR4/NF-κB信号通路相关靶标,调控炎性反应细胞因子的表达水平有关。

Objective To explore the effects of Atractylodes macrocephala-Paeonia lactiflora drug pair on the targets related to TLR4/NF-κB signalling pathway in the model rats of diarrhoeal irritable bowel syndrome(IBS-D)in an attempt to elucidate the potential mechanism of its therapeutic effect on the treatment of low-grade inflammatory response.Methods IBS-D model rats were established by senna gavage+chronic unpredictable stimulation,and randomly divided into model group,drug-pair group,Atractylodes macro⁃cephala group,Paeonia lactiflora group,and western medicine group,with 6 rats in each group.Take another 6 rats without modeling and set as the blank group.The rats in each drug group were gavaged with the corresponding drug solution and treated continuously for 14 d.The intervention effects of Atractylodes macrocephala and Paeonia lactiflora sinensis on the symptoms of the model rats were evaluated by body weight,water content of stool,and AWR scores;blood was taken from the abdominal aorta of the rats in each group,and tissues of the terminal part of the ileum were dissected,and the intervention effect of Atractylodes macrocephala and Paeonia lactiflora sinensis on the symptoms of inflammation as well as TLR4/NF-κB signalling pathway of the IBS-D rats was detected by qRT-PCR,HE staining,Elisa,and WB methods.NF-κB signalling pathway related targets regulation.Results Compared with the model group,the drug pair improved body weight,reduced stool water content(P<0.01),sugar-water preference(P<0.01),reduced AWR scores at 40 mm⁃Hg and 60 mmHg pressures(P<0.01),improved the inflammatory pathological state of ileocecal terminal tissues,down-regulated TLR4,NF-κB,and IL-1βmRNA expression(P<0.01),down-regulated serum TLR4,NF-κB,MyD88 expression,up-regu⁃lated serum IL-10 expression,and down-regulated MyD88,p-NF-κB p65,IL-6 protein expression(P<0.01).Conclusion The effect of Paeonia lactiflora on improving diarrhoea symptoms and visceral hypersensitivity in IBS-D rats may be related to the inhibition of the TLR4/NF-κB signalling pathway,which regulates the expression level of inflammatory response cytokines.