Mechanism of Chaihu Shugansan in Treatment of Functional Dyspepsia in Rats Based on Mitophagy and Pink1/Parkin Signaling Pathway
10.13422/j.cnki.syfjx.20221506
- VernacularTitle:基于线粒体自噬及Pink1/Parkin信号通路探讨柴胡疏肝散治疗功能性消化不良大鼠的作用机制
- Author:
Xiangxiang WANG
1
;
Yujiao WANG
1
;
Li LI
1
;
Qingling JIA
1
;
Kailin JIANG
1
;
Keming YANG
1
;
Jianghong LING
1
Author Information
1. Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine， Shanghai 201203，China
- Publication Type:Journal Article
- Keywords:
Chaihu Shugansan;
functional dyspepsia;
mitophagy;
PTEN-induced kinase 1 （Pink1）;
E3 ubiquitin ligase （Parkin）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2023;29(2):45-51
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism of Chaihu Shugansan in the treatment of functional dyspepsia in rats based on mitophagy and PTEN-induced kinase 1 （Pink1）/E3 ubiquitin ligase （Parkin）. signaling pathway. MethodAccording to the principle of random grouping， 40 SD rats were assigned into a normal group， a model group， a Chaihu Shugansan group， and a positive drug （domperidone） group， with 10 rats in each group. The rats in other groups except the normal group received tail-clamping stimulation to replicate the model of functional dyspepsia. After each time of stimulation， the rats in the normal， model， Chaihu Shugansan， and positive drug groups were administrated with normal saline， normal saline， Chaihu Shugansan （4.8 g·kg-1）， and an aqueous solution of domperidone （4.5 mg·kg-1）， respectively. After 28 days of modeling， the gastric emptying rate and the small intestine propulsion rate of the rats in each group were measured and the tissue samples were collected. Hematoxylin-eosin （HE） staining was employed for observation of damage in gastric antrum tissue， and transmission electron microscopy （TEM） for ultrastructural observation of gastric interstitial cells of Cajal （ICCs）. Immunofluorescence co-localization was adopted to observe the expression of cytochrome c oxidase （COX Ⅳ） and Parkin. Western blot was employed to determine the expression levels of microtubule-associated protein 1， light chain 3 （LC3）， and the mitophagy-associated proteins prohibitin2 （PHB2）， Pink1， Parkin， and ubiquitin-specific protease 30 （USP30）. ResultCompared with the normal group， the modeling decreased the gastric emptying rate and the small intestine propulsion rate （P<0.05）. Compared with the model group， Chaihu Shugansan increased the gastric emptying rate and the small intestine propulsion rate （P<0.05）. The results of TEM showed that Chaihu Shugansan reduced the swelling degree of mitochondria in gastric antrum tissue. Compared with the normal group， the modeling increased the fluorescence intensity of Parkin in mitochondria （P<0.01）， while such increase can be alleviated by Chaihu Shugansan （P<0.01）. Western blotting results showed that compared with the normal group， the modeling up-regulated the protein levels of LC3， Pink1， Parkin， and PHB2 （P<0.05， P<0.01） and down-regulated the protein level of USP30 （P<0.01）. Compared with the model group， Chaihu Shugansan down-regulated the protein levels of LC3， Pink1， Parkin， and PHB2 （P<0.05， P<0.01） and up-regulated the protein level of USP30 （P<0.01）. ConclusionChaihu Shugansan may treat functional dyspepsia by blocking the Pink1/Parkin signaling pathway to inhibit excessive mitochondrial autophagy in ICCs.
