碱性神经酰胺酶1在硫酸钠葡聚糖诱导小鼠溃疡性结肠炎中的保护作用

doi:10.3969/j.issn.1006-5725.2025.01.002

摘要/Abstract

摘要：

目的研究碱性神经酰胺酶1（Acer1）在溃疡性结肠炎（UC）的黏膜屏障和免疫调节中发挥的作用和影响。方法通过构建Acer1基因敲除鼠（Acer1 KO），利用硫酸钠葡聚糖（DSS）连续饮水7 d诱导构建UC模型，对比C57小鼠和Acer1 KO小鼠在UC疾病中症状严重程度，利用RT-PCR和免疫组化检测肠黏膜屏障（ZO-1、Occludin、Claudin、JAMA）的表达水平，通过ELISA评估全身免疫反应水平（IL-1β、IL-6、IL-23、IL-17、IL-10和TNF-α的水平）以及RT-PCR评估肠道炎症浸润程度（TNF-α、IL-1β、IL-6、IL-17、IL-21等）。结果自由饮水中C57和Acer1 KO小鼠两组无明显差异；在DSS诱导的UC模型中，与C57小鼠相比Acer1 KO小鼠存活率下降，体质量下降明显，机械屏障和黏液屏障蛋白表达程度显著降低，肠上皮屏障损伤严重，炎症反应强烈，细胞因子浸润明显，结果差异有统计学意义（P < 0.05）。结论 Acer1通过维持肠黏膜屏障完整性，防御内毒素抑制炎症浸润，在UC发生发展过程中具有延缓疾病进展的作用。

关键词: 碱性神经酰胺酶1, 溃疡性结肠炎, 机械屏障, 黏液屏障, 炎症损伤

Abstract:

Objective To study the role and influence of basic Alkaline ceramidase 1 on mucosal barrier and immune regulation in ulcerative colitis （UC）. Methods Acer1 knockout mice （Acer1 KO） were constructed， and the UC model was induced by continuous drinking water of sodium dextran sulfate （DSS） for 7 days， and the severity of symptoms of UC disease in C57 and Acer1 KO mice was compared. The expression levels of intestinal mucosal barriers （ZO-1， Occludin， Claudin， JAMA） were detected by RT-PCR and immunohistochemistry. Systemic immune response levels （IL-1b、IL-6、IL-23、IL-17、IL-10， and TNF-a） were assessed by ELISA and intestinal inflammatory infiltration levels （TNF-a、IL-1b、IL-6、IL-17、IL-21， etc.） were assessed by RT-PCR. Results There was no significant difference between C57 and Acer1 KO mice in free drinking water. In the DSS induced UC model， compared with C57 mice， the survival rate of Acer1 KO mice decreased， the weight decreased significantly， the mechanical barrier and mucus barrier protein expression levels decreased significantly， the intestinal epithelial barrier was seriously damaged， the inflammatory response was strong， and the cytokine infiltration was obvious， with statistical differences （P < 0.05）. Conclusion Acer1 can inhibit inflammatory infiltration by maintaining the integrity of intestinal mucosal barrier， preventing endotoxin， and delaying the progression of UC.

Key words: alkaline ceramidase 1, ulcerative colitis, mechanical barrier, mucus barrier, inflammatory injury

中图分类号:

R574.62

何榕茂,方泽扬,张芸芸,吴友谅,梁世秀,王斯琪. 碱性神经酰胺酶1在硫酸钠葡聚糖诱导小鼠溃疡性结肠炎中的保护作用[J]. 实用医学杂志, 2025, 41(1): 7-14.

Rongmao HE,Zeyang FANG,Yunyun ZHANG,Youliang WU,Shixiu LIANG,Siqi WANG. Protective effect of basic alkaline ceramidase 1 in ulcerative colitis[J]. The Journal of Practical Medicine, 2025, 41(1): 7-14.

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引物名称	正向序列	反向序列
GAPDH	GGAGAAACCTGCCAAGTATGA	TCCTCAGTGTAGCCCAAGA
ZO-1	CATCTCCAGTCCCTTACCTTTC	CCTCCAGGCTGACATTAGTTAC
Claudin	GGTTATCGGAACTGTGGTAGAA	GTGCTCAGGGAAGATGGTAAG
Occludin	CTGGATCTATGTACGGCTCACA	TCCACGTAGAGACCAGTACCT
JAMA	ACCAGTTGGCAAGAAGGTCACC	GTGAAGTTGTCCTGTGCCTACTC
MUC2	ATGCCCACCTCCTCAAAGAC	GTAGTTTCCGTTGGAACAGTGAA
Hesl	CCAGCCAGTGTCAACACGA	AATGCCGGGAGCTATCTTTCT
Math-1	GAGTGGGCTGAGGTAAAAGAGT	GGTCGGTGCTATCCAGGAG
SPDEF	AAGGCAGCATCAGGAGCAATG	CTGTCAATGACGGGACACTG
KLF4	GTGCCCCGZCTAACCGTTG	GTCGTTGAACTCCTCGGTCT
IL-6	TTTCCTCTGGTCTTCTGGAGTA	CTCTGAAGGACTCTGGCTTTG
IL-17	TTTAACTCCCTTGGCGCAAAA	CTTTCCCTCCGCATTGACAC
TNF-α	GTAGCCCACGTCGTAGCAAA	ACAAGGTACAACCCATCGGC
IL-1β	GAGGACATGAGCACCTTCTTT	GCCTGTAGTGCAGTTGTCTAA
IL-21	GCCAGATCGCCTCCTGATTA	CATGCTCACAGTGCCCCTTT
IL-23	ATGCTGGATTGCAGAGCAGTA	ACGGGGCACATTATTTTTATCT
CXCR1	TCTGGACTAATCCTGAGGGTG	GCCTGTTGGTTATTGGAACTCTC
CXCR2	ATGCCCTCTATTCTGCCAGAT	GTGCTCCGGTTGTATAAGATGAC

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