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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

31卷

期数:

2025年01期

页码:

36-40

栏目:

基础医学

出版日期:

2025-01-20

文章信息/Info

Title:

Experimental study on the protective effect of butyrate on inflammatory bowel disease

文章编号:

1006-8147(2025)01-0036-05

作者:

刘佳¹; 赵泽满¹; 2; 刘雅涵¹; 2; CA Cristopher¹; 矫政洧¹; 靳小石¹; 2

（1.河北大学临床医学院，保定071000；2.河北大学附属医院普通外科，保定071000）

Author(s):

LIU Jia¹; ZHAO Zeman¹; 2; LIU Yahan¹; 2; CA Cristopher¹; JIAO Zhengwei¹; JIN Xiaoshi¹; 2

（1.School of Clinical Medicine，Hebei University，Baoding 071000，China；2.General Surgery，Hebei University Affiliated Hospital，Baoding 071000，China）

关键词:

丁酸盐; 炎症性肠病; 脂多糖

Keywords:

butyrate;  inflammatory bowel disease;  lipopolysaccharide

分类号:

R333

DOI:

10.20135/j.issn.1006-8147.2025.01.0036

文献标志码:

A

摘要:

目的：探究丁酸盐（BT）在炎症性肠病（IBD）细胞模型中的保护作用。方法：将人正常结肠上皮细胞（FHC）进行培育，用脂多糖（LPS）诱导构建炎症性肠病细胞模型，分为对照组、炎症组及0.25 mmol/L BT+LPS组、0.5mmol/L BT+LPS组、1 mmol/L BT+LPS组。其中对照组正常培育，不需额外添加任何试剂，炎症组加入浓度为100 g/mL的LPS刺激FHC细胞24 h；BT+LPS组分别加入相应浓度（0.25、0.5、1 mmol/L）的BT干预FHC细胞24 h，再加入浓度为100 g/mL的LPS刺激24 h。使用不同浓度（0、0.5、1、2、5 mmol/L）的BT作用于FHC细胞24 h和48 h后，通过细胞活力检测确定BT的浓度范围；然后分别使用0.25、0.5及1 mmol/L的BT溶液干预FHC细胞24 h后，加入100 g/mL的LPS作用24 h，RT-qPCR和ELISA分别检测对照组、炎症组、0.25 mmol/L BT+LPS组、0.5 mmol/L BT+LPS组和1 mmol/L BT+LPS组中COX-2、白细胞介素（IL）-6 mRNA以及蛋白的水平，明确BT对FHC细胞的保护作用与浓度的关系；最后电镜观察对照组、炎症组及1 mmol/L BT+LPS组中细胞的微观结构变化。结果：BT的浓度在1 mmol/L以下时FHC细胞活力在90%以上，并且作用24 h和48 h后细胞活力无明显变化。RT-qPCR结果显示，与对照组相比，炎症组中COX-2 mRNA、IL-6 mRNA升高（均P<0.001）；与炎症组相比，1 mmol/L BT+LPS组COX-2 mRNA和IL-6 mRNA降低（t=3.384、4.722，均P<0.05）；0.5 mmol/L BT+LPS组COX-2 mRNA和IL-6 mRNA水平降低（t=2.817、3.753，均P<0.05）；0.25 mmol/L BT+LPS组COX-2 mRNA和IL-6 mRNA水平差异无统计学意义（均P>0.05）。与0.5 mmol/L BT+LPS组比较，1 mmol/L BT+LPS组COX-2 mRNA和IL-6 mRNA（t=4.561、5.196，均P<0.05）差异具有统计学意义。ELISA结果显示，与对照组相比，炎症组COX-2和IL-6表达水平升高（均P<0.001）；与炎症组相比，1 mmol/L BT+LPS组COX-2和IL-6水平降低（t=4.547、3.452，均P<0.001）；0.5 mmol/L BT+LPS组COX-2和L-6水平降低（t=2.927、3.265，均P<0.05）；0.25 mmol/L BT+LPS组COX-2和IL-6水平差异无统计学意义（均P>0.05）。与0.5 mmol/L BT+LPS组比较，1 mmol/L BT+LPS组IL-6和COX-2水平（t=4.674、3.217，均P<0.05）差异具有统计学意义。电镜下对照组FHC细胞展现出正常且完整的微观结构，实验组FHC细胞微绒毛变稀疏，紧密连接结构被破坏，1 mmol/L BT+LPS组FHC细胞微绒毛呈现部分脱落，紧密连接结构破坏减轻。结论：BT浓度在1 mmol/L以下时，对炎症性肠病细胞模型的保护作用随浓度的增加而增强。

Abstract:

Objective： To explore the protective effect of butyrate （BT） in inflammatory bowel disease （IBD） cell model. Methods：Human normal colon epithelial cells （FHC） were cultured and induced with lipopolysaccharide （LPS） to construct an inflammatory bowel disease cell model. The cells were divided into the following groups： control group， inflammation group， 0.25 mmol/L BT+LPS group， 0.5 mmol/L BT+LPS group， and 1 mmol/L BT+LPS group. The control group was cultured normally without the need for additional reagents， while the inflammation group was stimulated with LPS at a concentration of 100 g/mL for 24 hours to stimulate FHC cells. Different BT+LPS groups were treated with corresponding concentrations（0.25，0.5，1 mmol/L） of BT to intervene in FHC cells for 24 hours， followed by stimulation with LPS at a concentration of 100 μg/mL for 24 hours. After treating FHC cells with BT at different concentrations of 0， 0.5， 1， 2 and 5 mmol/L for 24 and 48 hours， the concentration range of BT was determined by cell viability detection. Then， 0.25， 0.5 and 1 mmol/L BT solutions were used to intervene in FHC cells for 24 hours. 100 g/mL LPS was added and treated for 24 hours. RT-qPCR and ELISA were used to detect the levels of COX-2， IL-6 mRNA and protein in the control group， inflammation group， 0.25 mmol/L BT+LPS group， 0.5 mmol/L BT+LPS group， and 1 mmol/L BT+LPS group， respectively， to clarify the protective effect of BT on FHC cells and its concentration relationship. Finally， the microstructural changes of cells in the control group， inflamma-tion group， and 1 mmol/L BT+LPS group were observed under electron microscopy. Results：When the concentration of BT was below 1 mmol/L， the viability of FHC cells was above 90%， and there was no significant change in cell viability after 24 and 48 hours of treatment. The RT-qPCR results showed that compared with the control group， COX-2 mRNA and IL-6 mRNA increased（both P<0.001） in the inflammation group. Compared with the inflammation group， the 1 mmol/L BT+LPS group showed a decrease in COX-2 mRNA and IL-6 mRNA（t=3.384， 4.722， both P<0.05）. The levels of COX-2 mRNA and IL-6 mRNA decreased in the 0.5 mmol/L BT+LPS group（t=2.817， 3.753， both P<0.05）. There was no statistically significant difference in the levels of COX-2 mRNA and IL-6 mRNA in the 0.25 mmol/LBT+LPS group（both P>0.05）. Compared with the 0.5 mmol/L BT+LPS group， the 1 mmol/L BT+LPS group showed statistically significant differences in COX-2 mRNA and IL-6 mRNA （t=4.561，5.196， both P<0.05）. The ELISA results showed that compared with the control group， the expression levels of COX-2 and IL-6 increased in the inflammatory group（both P<0.001）. Compared with the inflammation group， the levels of COX-2 and IL-6 were reduced in the 1 mmol/L BT+LPS group（t=4.547， 3.452， both P<0.001）. The levels of COX-2 and IL-6 decreased in the 0.5 mmol/L BT+LPS group（t=2.927，3.265， both P<0.05）. There was no statistically significant difference in the levels of COX-2 and IL-6 in the 0.25 mmol/L BT+LPS group （both P>0.05）. Compared with the 0.5 mmol/L BT+LPS group， the 1 mmol/L BT+LPS group showed statistically significant differences in IL-6 and COX-2 levels （t=4.674， 3.217， both P<0.05）. Under electron microscopy， the control group of FHC cells showed normal and intact microstructures. In the experimental group， the microvilli of FHC cells became sparse， and the tight junction structure was disrupted. In the 1 mmol/L BT+LPS group， the microvilli of FHC cells showed partial shedding， and the damage to the tight junction structure was reduced. Conclusion： The pro-tective effect of BT on inflammatory bowel disease cell models increases with increasing concentration when the concentration is below 1 mmol/L.

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备注/Memo

备注/Memo:

基金项目：河北大学医学学科培育项目（2023X03）
作者简介：刘佳（1997-），男，硕士在读，研究方向：外科学；通信作者：靳小石，E-mail：doctorjinxiaoshi@126.com。

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更新日期/Last Update: 2025-02-10