基于核因子κB信号通路观察清热化湿调枢方对溃疡性结肠炎小鼠的影响

Regulatory Effect of Qingre Huashi Tiaoshu Recipe on Mice with Ulcerative Colitis Based on NF-κB Signaling Pathway

目的:基于核因子κB(NF-κB)信号通路,探讨清热化湿调枢方对溃疡性结肠炎(UC)小鼠的作用及机制。方法:36只无特定病原体(SPF)级C57BL/6小鼠随机选取6只为正常组,其余小鼠均给予2.5%葡聚糖硫酸钠(DSS)水溶液建立UC小鼠模型。模型评价后,30只小鼠随机分为模型组、美沙拉嗪组、清热化湿调枢方高、中、低剂量组。后3组分别给予3.38、1.69、0.845 g/(kg·d)清热化湿调枢方混悬液0.5 mL灌胃;美沙拉嗪组给予0.52 g/(kg·d)美沙拉嗪混悬液0.5 mL灌胃;其余小鼠给予0.5 mL纯水灌胃,均1次/d,干预10 d。干预过程中测量小鼠体质量,干预结束后计算疾病活动指数(DAI),测量小鼠结肠长度,苏木精-伊红染色后观察远端结肠组织病理学形态,Western Blotting测定结肠核因子κB p65(NF-κB p65)和磷酸化NF-κB p65(p-NF-κB p65)水平,计算p-NF-κB p65/NF-κB p65值,实时荧光定量PCR(RT-qPCR)检测结肠白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)、趋化因子(KC/CXCL1)的mRNA水平。结果:与模型组比较,美沙拉嗪组、清热化湿调枢方高剂量组、中剂量组小鼠体质量增加、结肠长度一定程度恢复,结肠组织病理炎症减轻,干预前后DAI差值增加(均P<0.05);p-NF-κB p65、p-NF-κB p65/NF-κB p65值、IL-1β、KC/CXCL1、IL-6、TNF-α的mRNA水平降低(P<0.05)。结论:清热化湿调枢方能够改善UC小鼠症状、修复结肠黏膜,机制可能与抑制NF-κB信号通路激活,从而抑制炎症有关。

Objective:To explore the effect of Qingre Huashi Tiaoshu Recipe on mice with ulcerative colitis(UC)based on nuclear factor kappa B(NF-κB)signaling pathway and its mechanism.Methods:Six mice were randomly selected as the normal group from 36 SPF C57BL/6 mice.The remaining mice were given 2.5%dextran sulfate sodium(DSS)aqueous solution to establish a UC model.After evaluation of modeling,30 mice were randomly divided into a model group,a mesalazine group,and high,medium,and low-dose Qingre Huashi Tiaoshu Recipe groups.The latter three groups were given a daily dose of 3.38,1.69,and 0.845 g/(kg·d)of Qingre Huashi Tiaoshu Recipe suspension,respectively.The mesalazine group was given the daily dose of 0.52 g/(kg·d)mesalazine suspension,and the other two groups were given an equal volume of pure water.Each group was intervened continuously for 10 days.During intervention,the body weight of mice was measured.The disease activity index(DAI)after treatment was calculated.The colon length of mice was measured,and the histopathological changes in the distal colon obtained from mice were observed by hematoxylin-eosin(HE)staining.The expression levels of NF-κB p65 and p-NF-κB p65 in the colon was determined by Western blotting,and the value of p-NF-κB p65/NF-κB p65 was calculated.The mRNA levels of interleukin-1β(IL-1β),interleukin-6(IL-6),chemokine(KC/CXCL1),and tumor necrosis factor-α(TNF-α)were detected by real-time quantitative polymerase chain reaction(RT-qPCR).Results:After intervention,as compared with the model group,the body weight of mice in the mesalazine group and the high and medium-dose Qingre Huashi Tiaoshu Recipe groups was increased significantly,the length of the colon partly recovered,and the pathological inflammation of colon tissue was alleviated.The difference of DAI before and after intervention was increased(P<0.05).The values of p-NF-κB p65 and p-NF-κB p65/NF-κB p65 and the mRNA levels of IL-1β,KC/CXCL1,IL-6,and TNF-αin the mesalazine group and the high and medium-dose Qingre Huashi Tiaoshu Recipe groups were decreased(P<0.05).Conclusion:Qingre Huashi Tiaoshu Recipe can attenuate the symptoms of mice with UC and ameliorate intestinal mucosal impairment.The mechanism may be related to the inhibition of the activation of the NF-κB pathway,thereby inhibiting inflammation.