克痢痧对大鼠乙酸性结肠炎及炎症信号通路TLR4/NF-κB的影响研究

The effect of Kelisha on the inflammation in acetic acid-induced colitis rats and its research on TLR4/NF-κB signal pathway of the inflammatory response

目的:本实验通过观察克痢痧对大鼠乙酸性结肠炎的研究,揭示其对炎症信号通路TLR4/NF-κB的影响,为临床应用中成药防治溃疡性结肠炎提供实验依据。方法:70只大鼠随机分成正常组、模型组、6g/kg蒙脱石散组、0.35g/kg柳氮磺吡啶组、88.2、176.4、352mg/kg克痢痧组。除正常组外,其他六组采用10%乙酸溶液灌肠建立溃疡性结肠炎模型。造模完成后,七组正常饮食水,各组灌胃相应药物,连续14天。通过观察动物一般情况及大便情况,ELISA法测定血清IL-1β、IL-6、TNF-α、EGF含量;Western Blot法测TLR-4、NF-κB表达;并于实验第15天处死大鼠观察结肠粘膜损伤,评价损伤指数(CMDI)。结果:模型组大鼠结肠粘膜炎症明显高于正常组,0.352g/kg克痢痧组大鼠结肠粘膜炎症较模型组显著改善;造模后第14天352mg/kg克痢痧血清IL-1β、TNF-α含量较模型组显著降低;0.352g/kg克痢痧组EGF含量显著增高。在TLR4、NF-κB的表达上,克痢痧352mg/kg剂量组的表达显著低于模型组,且其表达量随药物剂量增加而降低。结论:克痢痧能使乙酸性结肠炎大鼠的一般状态及肠粘膜形态学得到较好改善;克痢痧能够降低乙酸性结肠炎大鼠血清IL-1β、IL-6、TNF-α水平,控制炎症;克痢痧可提高乙酸性结肠炎大鼠血清EGF水平,修复粘膜上皮;干预TLR4/NF-κB信号通路可能是克痢痧控制乙酸性结肠炎炎症反应的机制之一。

Objective： To study the curative effect of Kelisha through meauring inflammation factors,protective substances in rats＇ colonic mucosa and protein expressions of signaling pathway in acetic acid-induced colitis rats. Methods： 70 SD male rats were randomly divided into the normal group,the model group,6 g/kg Montmorillonite Powder group,350 mg/kg SASP group,88. 2 mg/kg Kelisha group,176. 4 mg/kg Kelisha group,352 mg/kg Kelisha group. Except the normal group,we administered 10% acetic acid into rats by clyster to establish the UC model. Saline was given by the gavage once a day to rats in the normal and model groups,and Montmorillonite Powder,SASP,different doses of Kelisha were given to rats in corresponding groups for 14 days. We performed the gross examination of colonic tissues,measured levels of serum IL-1β,IL-6,TNF-α,and EGF by ELISA and expressions of TLR-4 and NF-κB by Western Blot. On the 15 th day,rats were sacrificed and the colonic mucosa damage index（CMDI） was evaluated. Results： Compared with the control group,the inflammatory condition of the colon tissue in model group was more severe. Compared with the model group,the inflammatory condition of the colon tissue in 352 mg/kg Kelisha group was improved. Levels of serum IL-1β、IL-6 and TNF-αin 352 mg/kg Kelisha group were decreased. The level of serum EGF in 352 mg/kg Kelisha group was increased,and it was higher than that in SASP group,and the difference between 352 mg/kg Kelisha group and SASP group was statistically significant（P〈0. 05）. Expressions of TLR-4 and NFκB of Colon tissues in Kelisha group were decreased,and they decreased with the increase of doses. Conclusions： Kelisha can improve the colon mucosa condition of the acetic acid-induced colitis rats,inhibit the inflammatory response and intervene the TLR-4/NFκB signal pathway.