| 顾红,田玉雯,夏晶晶,等.中药灌肠方基于NLRP3/SIRT6信号通路改善葡聚糖硫酸钠诱导结肠炎的作用机制研究[J].浙江中医药大学学报,2024,48(11):1355-1362. |
| 中药灌肠方基于NLRP3/SIRT6信号通路改善葡聚糖硫酸钠诱导结肠炎的作用机制研究 |
| Herbal Enema Formula Ameliorates Dextran Sodium Sulfate-Induced Colitis by Modulating NLRP3/SIRT6 Signaling Pathway |
| DOI：10.16466/j.issn1005-5509.2024.11.004 |
| 中文关键词: 溃疡性结肠炎 中药灌肠方 炎症 氧化应激 肠道屏障 紧密连接 NLRP3 SIRT6 |
| 英文关键词: ulcerative colitis herbal enema formula inflammation oxidative stress intestinal barrier tight junction NLRP3 SIRT6 |
| 基金项目:江苏省中医药科技发展计划项目(YB2020046)；江阴市中医药学会科研项目(Z202202) |
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| 中文摘要: |
| [目的] 探究中药灌肠方对葡聚糖硫酸钠(dextran sulfate sodium，DSS)诱导的溃疡性结肠炎(ulcerative colitis，UC)的保护作用及其作用机制。[方法] 给予C57BL/6J小鼠自由饮用2.5%DSS水溶液7 d以诱导UC。2 d后给药组分别给予低、中、高剂量(1.3、2.6、5.2 ?g/kg)的中药灌肠方和阳性药柳氮磺吡啶0.2 ?g/kg灌肠，模型组和正常组给予等体积0.9%氯化钠溶液灌肠。以苏木精-伊红(hematoxylin-eosin，HE)染色观察小鼠结肠组织切片病变程度；采用酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)测定血清炎症因子肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白细胞介素-6(interleukin-6，IL-6)、白细胞介素-1β(interleukin-1β，IL-1β)及白细胞介素-18(interleukin-18，IL-18)水平；采用生理生化检测试剂盒检测髓过氧化物酶(myeloperoxidase，MPO)、超氧化物歧化酶(superoxide dismutase，SOD)活性，一氧化氮(nitric oxide，NO)、丙二醛(malondialdehyde，MDA)水平；免疫印迹法检测小鼠结肠组织闭锁小带蛋白-1(zonula occludens protein-1，ZO-1)、闭合蛋白(occludin)、核苷酸结合寡聚结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3，NLRP3)、沉默调节蛋白6(sirtuin 6，SIRT6)蛋白表达水平。[结果] 与正常组比较，DSS可诱导结肠炎症、隐窝丢失、杯状细胞耗竭、炎性浸润；血清TNF-α、IL-6、IL-1β及IL-18炎症因子水平，MPO活性和NO、MDA水平明显升高(P<0.01，P<0.001)，SOD活性明显降低(P<0.01)。与模型组比较，中药灌肠方治疗后明显改善DSS诱导的结肠炎症隐窝丢失、杯状细胞耗竭、炎性浸润的现象；血清TNF-α、IL-6、IL-1β及IL-18炎症因子水平，MPO活性和NO、MDA水平明显降低(P<0.01，P<0.001)，中剂量和高剂量组SOD活性明显升高(P<0.05)。中药灌肠方组小鼠结肠组织ZO-1、occludin、SIRT6的蛋白表达水平明显升高(P<0.001，P<0.01，P<0.05)，中剂量和高剂量组NLRP3蛋白表达水平明显降低(P<0.01)。[结论] 中药灌肠方对DSS诱导的UC具有治疗作用，其机制包括缓解炎症、减轻氧化应激、改善肠道屏障通透性以及调节NLRP3/SIRT6信号通路。 |
| 英文摘要: |
| [Objective] To investigate the protective effect of herbal enema formula on dextran sodium sulfate(DSS)-induced ulcerative colitis(UC) and its mechanism of action. [Methods] C57BL/6J mice were given 2.5% DSS aqueous solution for 7 d to induce UC. After 2 d， mice in herbal enema formula low， medium and high dose groups were given 1.3 ?g/kg，2.6 ?g/kg and 5.2 ?g/kg herbal enema formula for edema; 0.2 ?g/kg sulfasalazine were given to positive control group for enema; mice in model group and normal group were given equal volumes of 0.9% sodium chloride solution for enema， respectively. The degree of lesions was observed by hematoxylin-eosin(HE) staining in mice colon tissue sections; serum inflammatory factor tumor necrosis factor-α(TNF-α)， interleukin-6(IL-6)， interleukin-1β(IL-1β) and interleukin-18(IL-18) were determined by enzyme linked immunosorbent assay(ELISA); myeloperoxidase(MPO) and superoxide dismutase(SOD) activities， nitric oxide(NO) and malondialdehyde(MDA) levels were detected by physiological and biochemical assay kits; protein expression levels of zonula occludens protein-1(ZO-1)， occludin， nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)， and sirtuin 6(SIRT6) in mouse colon tissues were detected by western blot. [Results] Compared with normal group， DSS induced colonic inflammation， crypt loss， cuprocyte depletion and inflammatory infiltration; serum TNF-α， IL-6， IL-1β， and IL-18 inflammatory factor levels， MPO activity， NO and MDA levels were significantly increased(P<0.01， P<0.001)， and SOD activity was significantly decreased(P<0.01). After treatment， the herbal enema formula significantly improved the DSS-induced colonic inflammation of crypt loss， cuprocyte depletion， and inflammatory infiltration. The levels of serum TNF-α， IL-6， IL-1β and IL-18 inflammatory factors， MPO activity and NO and MDA levels were significantly decreased(P<0.01， P<0.001)， and SOD activity were significantly increased in medium dose and high dose groups(P<0.05). The protein expression levels of ZO-1， occludin and SIRT6 in the colonic tissues of mice in herbal enema formula group were significantly higher(P<0.001， P<0.01， P<0.05)， and the protein expression levels of NLRP3 in medium dose and high dose groups were significantly lower(P<0.01). [Conclusion] The herbal enema formula has a protective effect against DSS-induced UC， and its mechanism includes alleviating inflammation， reducing oxidative stress， improving intestinal barrier permeability， and regulating the NLRP3/SIRT6 signaling pathway. |
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