基于TLR4/NF-κB信号通路探讨黔产刺梨根干预溃疡性结肠炎的作用机制

陈波洋; 赵玲婕; 罗运凤; 管连城; 于海洋; 赵洁; 张旭飞; 蒋志滨; 陈云志

doi:10.13386/j.issn1002-0306.2021100024

基于TLR4/NF-κB信号通路探讨黔产刺梨根干预溃疡性结肠炎的作用机制

Study on the Mechanism of Guizhou Rosa roxburghii Radix on Ulcerative Colitis Based on TLR4/NF-κB Signaling Pathway

摘要

摘要: 目的：基于TLR4/NF-κB信号通路探讨黔产刺梨根治疗溃疡性结肠炎大鼠的作用机制。方法：采用2,4,6-三硝基苯磺酸（TNBS）/乙醇溶液灌肠建立溃疡性结肠炎模型，灌胃刺梨根水煎液高、中、低剂量组（8、4、2 g/kg），柳氮磺嘧啶组（0.3 g/kg）。观察大鼠外观、动作行为以及血便；采集大鼠血清与大鼠结肠，利用苏木素-伊红（HE）染色观察各组大鼠结肠病理学改变；酶联免疫吸附测定（ELISA）检测大鼠血清白细胞介素（IL）-1β，IL-6，TNF-α水平；逆转录聚合酶链式反应（RT-PCR）检测大鼠结肠Myd88、NF-κB p50、NF-κB p65、TLR4 mRNA表达；蛋白免疫印迹法（Western blot）检测大鼠结肠Myd88、NF-κB p50、NF-κB p65、TLR4蛋白表达。结果：刺梨根水煎液可明显改善溃疡性结肠炎大鼠结肠炎症损伤，特别是刺梨根水煎液高剂量组。与模型组比较，刺梨根水煎液高剂量组结肠病理损伤得到显著改善，刺梨根水煎液高剂量组血清IL-1β、IL-6、TNF-α水平均极显著下降（P<0.01）；结肠Myd88、NF-κB p50、TLR4 mRNA表达量显著下降（P<0.05）；结肠Myd88、NF-κB p50、NF-κB p65、TLR4蛋白表达量极显著下降（P<0.01）。结论：刺梨根水煎液可有效缓解TNBS诱导的溃疡性结肠炎并改善炎症损伤，刺梨根水煎液干预溃疡性结肠炎的作用机制可能与抑制TLR4/NF-κB信号通路相关。

Abstract: Objective: To explore the mechanism of Guizhou Rosa roxburghii Radix in the treatment of ulcerative colitis in rats based on TLR4/NF-κB signaling pathway. Methods: The ulcerative colitis model was established by gavage with 2,4,6-trinitrobenzenesulfonic acid (TNBS)/ethanol solution, gavage with Rosa roxburghii Radix aqueous decoction high, medium and low dose groups (8, 4 and 2 g/kg) and salazosulfanil group (0.3 g/kg). Observation of the appearance, motor behaviour and blood stool of the rats. Serum and colon of rats were collected, and hematoxylin-eosin (HE) staining was used to observe the pathological changes in the colon of each group of rats. Enzyme-linked immunosorbent assay (ELISA) for the detection of serum interleukin (IL)-1β, IL-6, TNF-α levels in rats. Reverse transcription-polymerase chain reaction (RT-PCR) for the detection of rat colon Myd88, NF-κB p50, NF-κB p65, TLR4 mRNA expression. Protein immunoblotting (Western blot) was performed to detect Myd88, NF-κB p50, NF-κB p65, TLR4 protein expression in rat colon. Results: Rosa roxburghii Radix aqueous decoction significantly improved colonic inflammatory injury in rats with ulcerative colitis, especially in the high dose group of Rosa roxburghii Radix aqueous decoction. Compared with the model group, the Rosa roxburghii Radix aqueous decoction high-dose group showed significant improvement in colon pathological damage. Serum IL-1β, IL-6 and TNF-α levels were significantly decreased in the Rosa roxburghii Radix aqueous decoction high-dose group (P<0.01); colon Myd88, NF-κB p50, TLR4 mRNA expression was significantly decreased (P<0.05); colon Myd88, NF-κB p50, NF-κB p65, TLR4 protein expression was significantly decreased (P<0.01). Conclusion: Rosa roxburghii Radix aqueous decoction can effectively alleviate TNBS-induced ulcerative colitis and ameliorate inflammatory damage. The mechanism of action of Rosa roxburghii Radix aqueous decoction intervention in ulcerative colitis may be related to the inhibition of TLR4/NF-κB signaling pathway.