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[WDSUB1基因敲低通过抑制核因子-κB信号通路减轻葡聚糖硫酸钠诱导的小鼠结肠炎]

[WDSUB1 knockdown alleviates dextran sulfate sodium-induced colitis in mice by inhibiting nuclear factor-κB signaling pathway].

作者信息

Wang S, Cui L, Liu X, Luo Z, Li H, Pu J

机构信息

Department of Gastroenterology, First Medical Center of Chinese PLA General Hospital, Beijing 100853, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2022 Aug 20;42(8):1119-1125. doi: 10.12122/j.issn.1673-4254.2022.08.02.

Abstract

OBJECTIVE

To explore the effect of WDSUB1 on dextran sulfate sodium (DSS)-induced inflammatory colon injury in mice and the underlying mechanism.

METHODS

Different WDSUB1 siRNA sequences were transfected into mouse fibroblast L929 cells and the optimal sequence was selected by Western blotting. Twelve male C57BL/6 mice were randomized into two groups for injection of siWDSUB1 or siControl via the caudal vein, followed by treatment with 2.5% DSS in drinking water to establish mouse models of DSS- induced colitis (=6). The expression level of WDSUB1 in the colon tissue of the mice was detected with Western blotting and RT-PCR, the changes in body weight and fecal condition were recorded, and the clinical symptoms of the mice were evaluated. The mRNA expression levels of IL-6, COX-2 and TNF-α and the protein expression of IκBα and P65 in the colon tissues were detected with RT- PCR and Western blotting, respectively.

RESULTS

The mRNA and protein expressions of WDSUB1 in the colon tissues were significantly lower in colitis mice with WDSUB1 knock-down than in the control mice. Compared with the control mice, the mice receiving siWDSUB1 injection showed obviously milder weight loss, diarrhea and hematochezia with significantly lower mRNA expressions of COX2, IL-6 and TNFα ( < 0.05) and protein expression of IκBα but without obvious changes in P65 expression in the colon tissue.

CONCLUSION

WDSUB1 knockdown can alleviate DSS- induced colitis in mice possibly by inhibiting the NF-κB signaling pathway and decreasing the expression of inflammatory factors in the colon tissues.

摘要

目的

探讨WDSUB1对葡聚糖硫酸钠(DSS)诱导的小鼠炎症性结肠损伤的影响及其潜在机制。

方法

将不同的WDSUB1 siRNA序列转染至小鼠成纤维细胞L929中,通过蛋白质免疫印迹法筛选出最佳序列。将12只雄性C57BL/6小鼠随机分为两组,经尾静脉注射siWDSUB1或siControl,随后饮用含2.5% DSS的水以建立DSS诱导的结肠炎小鼠模型(每组n = 6)。采用蛋白质免疫印迹法和RT-PCR检测小鼠结肠组织中WDSUB1的表达水平,记录体重和粪便状况的变化,并评估小鼠的临床症状。分别采用RT-PCR和蛋白质免疫印迹法检测结肠组织中IL-6、COX-2和TNF-α的mRNA表达水平以及IκBα和P65的蛋白质表达。

结果

WDSUB1敲低的结肠炎小鼠结肠组织中WDSUB1的mRNA和蛋白质表达明显低于对照小鼠。与对照小鼠相比,注射siWDSUB1的小鼠体重减轻、腹泻和便血明显较轻,结肠组织中COX2、IL-6和TNFα的mRNA表达显著降低(P < 0.05),IκBα的蛋白质表达降低,但P65表达无明显变化。

结论

敲低WDSUB1可能通过抑制NF-κB信号通路并降低结肠组织中炎症因子的表达来减轻DSS诱导的小鼠结肠炎。

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