芦丁对溃疡性结肠炎小鼠的干预作用及NLRP3炎症小体的影响

Intervention of rutin on mice with ulcerative colitis and its influence on NLRP3 inflammasome

目的观察芦丁对葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠的干预作用。探讨其对NLRP3炎症小体信号分子活化的影响。方法将60只SPF级C57BL/6J小鼠随机分为6组,每组10只。除空白组外,其余组均采用葡聚糖硫酸钠诱导建立溃疡性结肠炎模型,美沙拉嗪组同时给予美沙拉嗪缓释颗粒灌胃,芦丁高、中、低剂量组分别给予100 mg/(kg·d)、50 mg/(kg·d)、25 mg/(kg·d)芦丁溶液灌胃,空白组、模型组以等体积l%羧甲基纤维素钠溶液灌胃,均连续5 d。灌胃结束后,比较各组疾病活动度(DAI)评分、组织学评分,采用酶联免疫吸附法测定小鼠血清和结肠组织中白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)水平,采用qRT-PCR法检测结肠组织中NLRP3、IL-1βmRNA表达量,采用Western blot法检测结肠组织中NLRP3、Cleaved caspase-1、Cleaved IL-1β蛋白表达水平。结果与空白组比较,模型组小鼠DAI评分、组织学评分,血清及结肠组织中IL-1β和IL-18水平,结肠组织中NLRP3 mRNA表达量、IL-1βmRNA表达量和NLRP3、Cleaved caspase-1、Cleaved IL-1β蛋白表达水平均明显升高(P均<0.05);与模型组比较,芦丁高、中剂量组和美沙拉嗪组上述指标均明显降低(P均<0.05),且芦丁高剂量组血清及结肠组织中IL-1β和IL-18水平,结肠组织中NLRP3 mRNA表达量、IL-1βmRNA表达量和NLRP3、Cleaved caspase-1、Cleaved IL-1β蛋白表达水平均明显低于美沙拉嗪组(P均<0.05)。结论芦丁可明显改善溃疡性结肠炎小鼠临床症状,抑制黏膜炎症损伤,其作用机制可能与抑制NLRP3炎症小体的活化有关。

Objective It is to observe the intervention of rutin on mice with ulcerative colitis induced by sodium dextran sulfate,and to explore its effect on the activation of NLRP3 inflammasome signaling molecules.Methods Sixty SPF C57BL/6J mice were randomly divided into 6 groups with 10 mice in each group.Except for the blank group,all the other groups were induced to establish ulcerative colitis models with dextran sodium sulfate.The mesalazine group was given mesalazine sustained-release granules by gavage at the same time,and the rutin high,medium and low dose groups were respectively given 100 mg/(kg·d),50 mg/(kg·d),25 mg/(kg·d)rutin solution by gavage,the blank group and the model group were given an equal volume of l%sodium carboxymethylcellulose solution by gavage,all groups were treated for 5 consecutive days.After gavage,the disease activity(DAI)scores and histological scores of each group were compared,and the levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)in serum and colon tissue of the mice were determined by enzyme-linked immunosorbent assay,the expression of NLRP3,IL-1β mRNA in colon tissue was detected by qRT-PCR method,the expression of NLRP3,Cleaved caspase-1,Cleaved IL-1β protein in colon tissue were detected by Western blot method.Results Compared with the blank group,the DAI score,histological score,the levels of IL-1β and IL-18 in serum and colon tissue,NLRP3 mRNA expression,IL-1β mRNA expression and the levels of NLRP3,Cleaved caspase-1,Cleaved IL-1β protein expression in colon tissue of the mice in the model group were significantly increased(all P<0.05);compared with the model group,the above indicators of rutin high,medium dose groups and mesalazine group were significantly reduced(P<0.05),and the levels of IL-1β and IL-18 in serum and colon tissue,NLRP3 mRNA expression,IL-1β mRNA expression and the levels of NLRP3,Cleaved caspase-1,Cleaved IL-1β protein expression in colon tissue of the rutin high-dose group were significantly lower than those of the mesalazine group(all P<0.05).Conclusion Rutin can significantly improve the clinical symptoms of mice with ulcerative colitis and inhibit mucosal inflammation.Its mechanism may be related to the inhibition of NLRP3 inflammasome activation.