ABSTRACT
ObjectiveTo investigate the effect of Zhizi Dahuang decoction （ZZDHT） in the treatment of alcoholic liver disease （ALD） by improving oxidative stress in hepatic neutrophils. MethodsNetwork pharmacology was used to obtain the chemical components of ZZDHT and their corresponding action targets and analyze the potential targets and functional pathways of ZZDHT in the treatment of ALD. The non-target metabolomics technology was used to observe the changes in the metabolites of ZZDHT in mouse serum and liver. The mice were given ZZDHT at a dose twice as much as the middle dose concentration by gavage， and serum and liver samples were collected at six time points after gavage （10 minutes， 30 minutes， 1 hour， 2 hours， 4 hours， and 6 hours） and were then mixed for mass spectrometry （administration group with 18 mice）， while the 18 mice in the control group were given an equal volume of normal saline by gavage. Ultra-performance liquid chromatography was used for rapid isolation and identification of the metabolites of ZZDHT in serum and liver tissue， and the effective constituents of ZZDHT were validated. Male C57BL/6J mice， aged 8 weeks， were randomly and equally divided into control group， model group， and low-， middle-， and high-dose ZZDHT groups， with 10 mice in each group. All mice except those in the control group were used to establish a mouse model of ALD （NIAAA model mice）， and at the same time， the mice in the administration groups were given low-， middle-， and high-dose ZZDHT by gavage， while those in the control group and the model group were given an equal volume of normal saline by gavage. The serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and triglyceride （TG） were measured； PCR was used to measure the gene expression levels of related inflammation， oxidative stress， and neutrophil indicators in the liver； ELISA was used to measure the levels of related inflammation and oxidative stress indicators in serum； superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and malondialdehyde （MDA） were measured to observe the level of oxidative stress in the liver； HE staining， myeloperoxidase staining， and oil red staining were used to observe liver injury， neutrophil infiltration， and lipid deposition. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsA total of 53 active components and 227 target genes were obtained for ZZDHT， and there were 8685 target genes of ALD， resulting in 222 common target genes between these two groups of genes. Core pathways included the interleukin-6 signaling pathway and the TNF signaling pathway. The non-targeted metabolic analysis of ZZDHT obtained 225 metabolites in mouse liver and 227 metabolites in serum， among which there were 126 common metabolites. The core pathways of liver metabolites included glycerolipid metabolism and inflammatory mediator regulation of TRP channels， and the core pathways of serum metabolites included the AMPK signaling pathway and oxidative phosphorylation， all of which were associated with oxidative stress- and inflammation-related pathways. Compared with the model group， the low-， middle-， and high-dose ZZDHT groups had significant reductions in the serum levels of ALT， AST， and TG （all P<0.05）， and the middle-dose ZZDHT group had significant reductions in the levels of Ly6g， Ncf1， Ncf2， IL-6， TNF-α， IL-1β， MDA， 4-HNE， Gp91， and P22 in the liver （all P<0.05）， a significant increase in the level of SOD （P<0.05）， a significant reduction in the serum level of 4-HNE （P<0.05）， and a significant increase in the level of GSH-Px （P<0.05）. There were significant improvements in fat deposition and neutrophil infiltration in the liver of mice in the middle-dose ZZDHT group （both P<0.05）. ConclusionZZDHT significantly reduces oxidative stress and inflammatory response in NIAAA model mice.
ABSTRACT
Liquid-assisted surface desorption atmospheric pressure chemical ionization mass spectrometry (LA-DAPCI-MS)was used directly for the analysis of healthy rat liver (normal control group);acute alcoholic injury rat liver(alcohol model group)and Zhizi Dahuang decoction-treated rat liver (ZZDHD group)sections with the mass spectrometry images obtained simultaneously.Principal component analysis (PCA)was employed for the differentiation of livers of the groups;and 3 phospholipids;PC (34 ∶2);PC(36 ∶4)and PC(38 ∶4);were found to be the main factors.LA-DAPCI mass spectrometry images showed that the 3 phospholipids were evenly distrib-uted in liver under the spatial resolution of 0.01 mm2.Contents of 3 PCs in the model group were decreased in alcohol model group compared to the normal control group and the ZZDHD group;which revealed the relationship between acute alcoholic liver injury and intrahepatic phospholipid variation.The results showed that ZZDHD had protective effect on acute alcoholic liver injury-induced intrahepatic phospholipid variations at the molecular level.
ABSTRACT
Objective: To study the blood components in Zhizi Dahuang Decoction (ZDD) and to further explore its pharmacodynamic material basis. Methods: UFLC fingerprints of serum in rats after ZDD treatment were established with serum pharmacochemistry method. The UFLC profiles of serum samples collected after the treatment of ZDD, ZDD without one of the component herbs and single crude herb were compared, and the transitional constituents to blood and their metabolites were analyzed. Results: Twenty-one constituents absorbed into rat blood were detected after oral administration of ZDD, 12 of which were metabolites. Other nine were prototype constituents, three of which were recognized as geniposide, aloeemodin, and rhein by comparing with reference substances, respectively. Conclusion: The constituents of ZDD absorbed into blood are preliminarily identified, which could lay the foundation for the illumination of its pharmacodynamic material basis.