1.Liver histopathological features of patients with drug-induced liver injury due to Chinese herbal medicine or Western medicine: A comparative analysis of 50 cases
Qiong LUO ; Gerui ZHU ; Hongtu GU
Journal of Clinical Hepatology 2020;36(3):596-601
ObjectiveTo compare the liver histopathological features of drug-induced liver injury (DILI) caused by Chinese herbal medicine (HM) and that caused by Western medicine (WM), and to investigate the pathological features of HM-DILI. MethodsA total of 50 patients with drug-induced liver injury(DILI) who were diagnosed and treated in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from March 2014 to June 2019. All the patients with DILI were enrolled and divided into HM-DILI group and WM-DILI group. A retrospective analysis was performed for the clinical features and the pathological features of the two groups, including the clinical type of DILI, severity of liver injury, and liver histopathological features, especially location and grade of liver inflammation, type and degree of steatosis, and iron deposition. The t test and Mann-Whitney U test was used for comparison of continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups; the Mann-Whitney U test was used for comparison of ranked data. ResultsOf all 50 patients, 20 (40.0%) had HM-DILI and 30 (60%) had WM-DILI. There were no significant differences in sex, age, course of disease, and biochemical parameters of liver function between the two groups (all P>0.05). The main clinical types of the two groups were hepatocellular injury type and the degree of injury which was mainly grade l, and there were no significant differences in the clinical type of DILI and the severity of liver injury between the two groups (both P>0.05). As for liver histopathological features, both groups had varying degrees of focal necrosis of hepatocytes, steatosis (microvesicular, macrovesicular, and mixed), and fibrous tissue proliferation at the portal area, and compared with the WM-DILI group, the HM-DILI group had significantly greater lymphocyte/plasma cell infiltration in the portal area (χ2=3.860, P<0.05) and iron deposition in Kupffer cells (χ2=4787, P<0.05). ConclusionThere are no significant differences in clinical manifestations between HM-DILI and WM-DILI, but patients with HM-DILI have more obvious inflammation in the portal area and iron deposition in Kupffer cells than those with WM-DILI.
2.A transcriptomic analysis of acute hepatotoxicity induced by aristolochic acid Ⅰ in mice
Gerui ZHU ; Jing WANG ; Kai HUANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Journal of Clinical Hepatology 2021;37(10):2389-2394
Objective To investigate the molecular mechanism of aristolochic acid Ⅰ (AAⅠ) inducing acute hepatotoxicity in mice. Methods A total of 15 male C57BL/6 mice were randomly divided into normal group with 6 mice and treatment group with 9 mice. The mice in the treatment group were given intraperitoneal injection of AAⅠ at a dose of 20 mg/kg for 5 consecutive days and were sacrificed to collect samples on day 6. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured, and HE staining was used to observe liver histological changes; three liver tissue samples were randomly selected from each group, and RNA was extracted for high-throughput transcriptome sequencing. Bioinformatics analysis and functional prediction were used to screen out differentially expressed genes, and quantitative real-time PCR (qRT-PCR) was used for validation. The t -test was used for comparison of continuous data between two groups. Results Compared with the normal group, the treatment group had significant increases in the activities of ALT and AST ( t =4.331 and 4.947, both P < 0.01), as well as disordered structure of hepatic lobules and spotted necrosis in hepatic lobules shown by HE staining. A total of 1352 differentially expressed genes were obtained based on the screening conditions of logFC > 2 and P < 0.05, among which there were 703 upregulated genes and 649 downregulated genes. The GO and KEGG enrichment analyses of these differentially expressed genes showed significant enrichment in GO terms (such as small molecular catabolism, immune response involving neutrophils, cytoplasmic vesicle lumen in secretory granules, cytoplasmic vesicle lumen, extracellular structural organization, and extracellular matrix) and KEGG pathways (such as chemical carcinogenesis, retinol metabolism, arachidonic acid metabolism, steroid hormone biosynthesis, transcriptional dysregulation in cancer, protein digestion and absorption, regulation of TRP channel by inflammatory mediators, drug metabolism, complement and coagulation cascade, glutathione metabolism, and the PPAR signaling pathway). A cluster analysis ( P < 0.05) showed that significantly downregulated genes included Srd5a1, Lipc, Aqp8, Hba-a1, Slco1a1, and Pklr, which were validated by qRT-PCR (all P < 0.05). Conclusion AA Ⅰ can lead to significant acute hepatotoxicity, which mainly involves the processes such as chemical carcinogenesis, retinol metabolism, arachidonic acid metabolism, steroid hormone biosynthesis, and transcriptional dysregulation in cancer.
3.Role of glutathione transferase in nonalcoholic fatty liver disease: An analysis based on gene expression profile
Tingting SHEN ; Gerui ZHU ; Fan WANG ; Xin SUN ; Kai HUANG ; Yuan PENG ; Yanyan TAO ; Chenghai LIU
Journal of Clinical Hepatology 2023;39(1):89-96
Objective To investigate the role of glutathione transferase in nonalcoholic fatty liver disease (NAFLD) induced by high-fat diet using the RNA-Seq technique in combination with gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of differentially expressed genes. Methods A total of 14 male C57BL/6J mice were divided into control group with 6 mice and model group with 8 mice by random sampling. The mice in the control group were fed with normal diet, and those in the model group were fed with high-fat diet for 7 consecutive weeks to establish a model of NAFLD. Kits were used to measure the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the level of triglyceride (TG), and HE staining and oil red staining were used to observe liver pathology and deposition of lipid droplets. Liver tissue RNA was extracted for RNA-Seq, and genes with a fold change of ≥2.0 and a P value of < 0.05 were defined as differentially expressed genes; after differentially expressed genes were screened out between the control group and the model group, GO and KEGG enrichment analyses were performed, and qRT-PCR was used to validate the expression of the differentially expressed genes. The independent samples t -test was used for comparison of normally distributed continuous data between two groups. Results There were no significant differences between the two groups in body weight and the serum levels of ALT and AST (all P > 0.05). Compared with the control group, the model group had a significantly higher serum level of TG (2.02±0.50 mmol/L vs 1.00±0.29 mmol/L, t =-4.45, P =0.001). HE staining showed diffuse steatosis and ballooning degeneration in the model group, and oil red staining showed that the model group had a significant increase in orange-red lipid droplets in the cytoplasm of hepatocytes and a significantly higher grade of hepatocyte steatosis than the control group (1.88±0.64 vs 1.00±0.00, t =-3.86, P =0.006). RNA-seq results showed a total of 1367 differentially expressed genes between the two groups, among which there were 608 upregulated genes and 759 downregulated genes, and there were 17 differentially expressed GST genes between the two groups. The top 10 GST genes in terms of fold change were validated, and compared with the control group, the model group had downregulated expression of GSTa2, GSTa3, GSTa4, GSTm1, GSTm2, GSTm3, GSTm4, GSTp1, and GSTo1 and upregulated expression of GSTk1. The results of qRT-PCR were consistent with the results of sequencing. Conclusion GST affects lipid metabolism by participating in various biological processes such as steroid metabolism, fatty acid metabolism, and cholesterol metabolism and is closely associated with the pathogenesis of NAFLD.