Naringin alleviates intestinal mucosal injury in mice with intra-abdominal infection by inhibiting non-canonical pyroptosis through P2X7 receptor

Yuan MA; Qianwen DUAN; Xupeng DONG; Che LIU; Ming ZHANG; Yuqing MA

doi:10.16016/j.2097-0927.202307058

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Naringin alleviates intestinal mucosal injury in mice with intra-abdominal infection by inhibiting non-canonical pyroptosis through P2X7 receptor

Basic Medicine | 更新时间：2026-01-09

- Naringin alleviates intestinal mucosal injury in mice with intra-abdominal infection by inhibiting non-canonical pyroptosis through P2X7 receptor
- Journal of Army Medical University Vol. 46, Issue 2, Pages: 139-146(2024)
- 作者机构：
  
  1.730000 兰州，兰州大学第一临床医学院
  2.730000 兰州，兰州大学第一医院麻醉科
- 作者简介：
- 基金信息：
  
  the Natural Science Foundation of Gansu Province(21JRIRA077)
- DOI：10.16016/j.2097-0927.202307058
  CLC： R282.71;R285.5;R574
- Received：11 July 2023，
  
  Revised：2023-09-06，
  
  Published：30 January 2024
- 稿件说明：
移动端阅览
Yuan MA, Qianwen DUAN, Xupeng DONG, et al. Naringin alleviates intestinal mucosal injury in mice with intra-abdominal infection by inhibiting non-canonical pyroptosis through P2X7 receptor[J]. Journal of Army Medical University, 2024, 46(2): 139-146.
DOI：

Yuan MA, Qianwen DUAN, Xupeng DONG, et al. Naringin alleviates intestinal mucosal injury in mice with intra-abdominal infection by inhibiting non-canonical pyroptosis through P2X7 receptor[J]. Journal of Army Medical University, 2024, 46(2): 139-146. DOI： 10.16016/j.2097-0927.202307058.

摘要

目的

探究柚皮苷(naringin

Ng)在腹腔感染(intra-abdominal infection

IAI)小鼠肠黏膜损伤中的治疗作用及其机制。

方法

腹腔注射脂多糖(lipopolysaccharide

LPS)建立IAI小鼠肠黏膜损伤模型。64只健康雄性小鼠采用随机数字表法分为4组(

=16)：对照组(Control组，0.2 mL PBS)，脂多糖组(LPS组，LPS 10 mg/kg)，柚皮苷组(Ng组，Ng 50 mg/kg+ LPS 10 mg/kg)和BzATP组(BzATP 5 mg/kg+Ng 50 mg/kg+LPS 10 mg/kg)。Ng和BzATP在造模前1 h进行腹腔注射，Control组予以等量PBS。各组于造模24 h随机选6只小鼠获取回肠。H

E染色观察回肠黏膜病理学变化和Chiu’s评分; 免疫荧光观察肠黏膜P2X7、焦亡效应蛋白GSDMD、巨噬细胞标记物CD68表达和分布情况；Western blot检测回肠组织P2X7和焦亡效应蛋白NLRP3、Caspase11、GSDMD及GSDMD-N表达量；ELISA检测回肠炎症因子IL-1β、IL-18和IL-10水平。记录7 d内小鼠脓毒症评分(mouse sepsis score，MSS)。

结果

LPS组与Control组相比，MSS和Chiu’s评分增高(

0.05)，回肠黏膜炎性损伤；肠黏膜P2X7、GSDMD和CD68免疫荧光分布一致且信号增强(

0.05)；促炎因子IL-1β、IL-18水平上升，抗炎因子IL-10水平下降(

0.05)；回肠组织P2X7、NLRP3、Caspase11、GSDMD和GSDMD-N蛋白表达增加(

0.05)。Ng组与LPS组相比，MSS下降；HE染色和Chiu’s评分表明肠黏膜炎性损伤减轻；P2X7、GSDMD、CD68荧光信号减弱(

0.05)；IL-1β、IL-18水平下降(

0.05)，IL-10水平上升(

0.05)；P2X7、NLRP3、Caspase11、GSDMD和GSDMD-N蛋白表达下降(

0.05)。BzATP组实验结果与Ng组相反。

结论

IAI小鼠肠黏膜P2X7受体表达升高，Caspase11介导的非经典焦亡发生，加重其肠黏膜损伤；Ng可能通过抑制P2X7受体调控非经典焦亡，改善IAI小鼠肠黏膜损伤。

Abstract

Objective

To investigate the therapeutic efficacy and underlying mechanism of naringin (Ng) for intestinal mucosal injury in mice with intra-abdominal infection (IAI).

Methods

A mouse IAI model was established by intraperitoneal injection (i.p.) of lipopolysaccharide (LPS). Sixty-four healthy male mice were randomly assigned to 4 groups (

=16): Control group (0.2 mL PBS)

LPS group (LPS 10 mg/kg)

Ng group (50 mg/kg Ng+10 mg/kg LPS)

and BzATP group (5 mg/kg BzATP +50 mg/kg Ng+10 mg/kg LPS). Ng and BzATP were administered i.p. 1 h before modeling

while the Control group received an equivalent volume of PBS. At 24 h post-modeling

6 mice from each group were randomly selected and sacrificed for ileum collection

and the histopathological changes in the ileal mucosa and Chiu's score were assessed using HE staining. Immunofluorescence assay was employed to investigate the expression and distribution of intestinal mucosal P2X7

pyroptosis effector protein GSDMD

and macrophage mark

er CD68. Western blotting was conducted to detect the expression of P2X7 and pyroptosis-related proteins NLRP3

Caspase11

GSDMD

and GSDMD-N protein in ileum tissue. ELISA was utilized to measure the contents of inflammatory factors IL-1β

IL-18

and IL-10 in the ileum. The mouse sepsis score (MSS) was recorded within 7 d.

Results

In comparison to the Control group

the LPS group exhibited significantly higher MSS and Chiu's scores (

0.05)

along with inflammation-associated damage to the ileal mucosa; Immunofluorescence assay displayed consistent and intensified distribution of intestinal mucosal P2X7

GSDMD

and CD68 (

0.05); Levels of pro-inflammatory factors IL-1β and IL-18 were increased

while that of anti-inflammatory factor IL-10 was decreased (

0.05); Additionally

the protein expression of P2X7

NLRP3

Caspase11

GSDMD

and GSDMD-N in ileum tissue was increased (

0.05). In contrast to the LPS group

the Ng group displayed a decrease in MSS; HE staining and Chiu's scores indicated a reduction in inflammatory damage to the intestinal mucosa; Moreover

fluorescence signals for P2X7

GSDMD

and CD68 were decreased (

0.05). Levels of IL-1β and IL-18 were decreased (

0.05)

while that of IL-10 was elevated (

0.05). The expression of P2X7

NLRP3

Caspase11

GSDMD

and GSDMD-N protein was decreased (

0.05). The results of the BzATP group were in contrast to those of the Ng group.

Conclusion

Elevoted expression of P2X7 receptor and Caspase11-mediated non-canonical pyroptosis aggravates the intestinal mucosal injury IAI mice. Ng may ameliorate the injury in IAI mice by suppressing P2X7 receptor and modulating non-canonical pyroptosis.

关键词

Keywords

references

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