Mechanism of Qinggan Jianpi Huoxue Prescription Against Hepatic Fibrosis via FoxO1 Mediated Regulation of Glycolytic Metabolic Reprogramming of Hepatic Stellate Cells
10.13422/j.cnki.syfjx.20260766
- VernacularTitle:清肝健脾活血方通过FoxO1调控肝星状细胞糖酵解代谢重编程抗肝纤维化机制
- Author:
Fuzhen PAN
1
;
Meng ZHU
2
;
Chuanjian SHI
1
;
Shiqiang XU
1
;
Ding LIU
3
;
Jinfang ZHANG
1
Author Information
1. Shenzhen Hospital （Futian） of Guangzhou University of Chinese Medicine，Shenzhen 518000，China
2. School of Basic Medical Sciences，Zhejiang Chinese Medical University，Hangzhou 310000，China
3. Jiangmen Wuyi Hospital of Traditional Chinese Medicine，Jiangmen 529000，China
- Publication Type:Journal Article
- Keywords:
Qinggan Jianpi Huoxue prescription;
hepatic fibrosis;
metabolomics;
hepatic stellate cells;
glycolysis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(18):98-107
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism of Qinggan Jianpi Houxue prescription against hepatic fibrosis in mice using metabolomics. MethodsA hepatic fibrosis mouse model was established by intraperitoneal injection of 20% carbon tetrachloride （CCl4， diluted with olive oil at a ratio of 4∶1） at a dose of 2.5 mL·kg-1 for 4 weeks. A total of 54 C57BL/6J mice were randomly divided into the blank control group， model group， Qinggan Jianpi Houxue prescription high-dose group （47.32 g·kg-1）， medium-dose group （23.66 g·kg-1）， low-dose group （11.83 g·kg-1）， and Biejiajian pill group （1.365 g·kg-1）， with 9 mice in each group. Concurrently with modeling， each administration group received the corresponding dose continuously for 4 weeks， once daily. After 4 weeks， liver tissues and blood samples were collected for liver function and hepatic pathological examinations. Non-targeted metabolomics was employed to detect hepatic metabolites. Principal component analysis （PCA）， partial least squares discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were performed to explore the metabolites and their involved metabolic pathways in the liver tissues of mice in the blank control group， model group and Qinggan Jianpi Houxue prescription high-dose group. Western blot validation was conducted at both the cellular and animal levels. ResultsCompared with the blank control group， the model group showed significantly increased levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） （P<0.01）， along with increased inflammatory cell infiltration， and formation of fibrous septa. The predominantly enriched pathway was the forkhead box O （FoxO） signaling pathway. Compared with the model group， the Qinggan Jianpi Huoxue prescription high-dose group exhibited significantly decreased ALT and AST levels （P<0.01）. In all administration groups of Qinggan Jianpi Huoxue prescription， inflammatory cell infiltration was reduced， fibrous septa were narrowed and diminished， collagen fibers were reduced， and the collagen area was significantly decreased. Non-targeted metabolomics identified 79 differential metabolites， among which 49 were up-regulated and 30 down-regulated， primarily enriched in the tricarboxylic acid （TCA） cycle and pyruvate metabolism pathways. Western blot results demonstrated that， compared with the blank control group， the model group exhibited significantly upregulated protein expressions of α-smooth muscle actin （α-SMA）， lactate dehydrogenase A （LDHA）， FoxO1， pyruvate kinase M2 （PKM2）， hexokinase 2 （HK2）， and phosphofructokinase 1 （PFK1） in liver tissues （P<0.05， P<0.01）. In hepatic stellate cells， the glucose concentration increased， and the protein expression of α-SMA， LDHA， FoxO1， PKM2， glucose transporter 1 （GLUT1）， and PFK1 were significantly up-regulated （P<0.05， P<0.01）. Conversely， compared with the model group， the Qinggan Jianpi Huoxue prescription high-dose group showed significantly downregulated protein expression of α-SMA， LDHA， FoxO1， PKM2， HK2， and PFK1 in liver tissues （P<0.05， P<0.01）. In hepatic stellate cells， the glucose concentration decreased， and the protein expression of α-SMA， LDHA， FoxO1， PKM2， GLUT1， and PFK1 were significantly down-regulated （P<0.05， P<0.01）. ConclusionQinggan Jianpi Huoxue prescription can alleviate the pathological progression of hepatic fibrosis in mice， potentially by regulating the TCA cycle and pyruvate metabolism through FoxO1， and by modulating glycolytic metabolic reprogramming in hepatic stellate cells， thereby promoting liver tissue repair.
