Modified Lianpoyin Formula Treats Hp-associated Gastritis by Regulating Mitochondrial Autophagy and NLRP3 Inflammasome Signaling Pathway
10.13422/j.cnki.syfjx.20251297
- VernacularTitle:基于线粒体自噬及NLRP3炎症小体信号通路探讨连朴饮加味方治疗Hp相关性胃炎小鼠的作用机制
- Author:
Siyi ZHANG
1
;
Haopeng DANG
1
;
Wenliang LYU
1
;
Wentao ZHOU
2
;
Wei GUO
2
;
Lin LIU
1
;
Lan ZENG
1
;
Yujie SUN
1
;
Luming LIANG
1
;
Yi ZHAO
1
Author Information
1. College of Chinese Medicine，Hubei University of Chinese Medicine，Wuhan 430000，China
2. Wuxue City Hospital of Traditional Chinese Medicine，Wuxue 435400，China
- Publication Type:Journal Article
- Keywords:
Helicobacter pylori;
gastritis;
modified Lianpoyin formula;
mitochondrial autophagy;
NOD like receptor protein 3（NLRP3） inflammasome
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(21):178-187
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg-1·d-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg-1·d-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg-1·d-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg-1·d-1） + amoxicillin （303 mg·kg-1·d-1） + clarithromycin （151.67 mg·kg-1·d-1） + colloidal pectin capsules （30.3 mg·kg-1·d-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.
