OBJECTIVES: To explore the therapeutic mechanism of compound Centella asiatica formula (CCA) for alleviating Schistosoma japonicum (Sj)-induced liver fibrosis in mice. METHODS: The active components and targets of CCA were identified using the TCMSP database with cross-analysis of Sj-related liver fibrosis targets. A "drug-component-target-pathway-disease" network was constructed using Cytoscape 3.9.1. Functional enrichment analysis (GO/KEGG) was performed using DAVID. Molecular docking study was carried out to validate interactions between the core targets and the key compounds. For experimental validation of the results, 36 mice were divided into control group, Sj-infected model group, and CCA-treated groups. In the latter two groups, liver fibrosis was induced via abdominal infection with Sj cercariae for 8 weeks, followed by 8 weeks of daily treatment with CCA decoction or saline. Hepatic pathology of the mice was assessedwith HE and Masson staining, and hepatic expressions of collagen-I and collagen-III were detected using immunohistochemistry; serum IL-6 and TNF-α levels were determined with ELISA. Hepatic expressions of TLR4 and MyD88 proteins were analyzed with Western blotting. RESULTS: We identified a total of 107 bioactive CCA components and 791 targets, including 37 intersection targets linked to Sj-induced fibrosis. The core targets included TNF, TP53, JUN, MMP9, and CXCL8, involving the IL-17 signaling, lipid metabolism, TLR4/MyD88 axis, and cancer pathways. Molecular docking study confirmed strong binding affinity between quercetin (a primary CCA component) and TNF/TP53/JUN/MMP9. In Sj-infected mouse models, CCA treatment significantly attenuated hepatic inflammatory cell infiltration, reduced collagen-I and collagen-III deposition, improved tissue architecture, reduced serum IL-6 and TNF-α levels, and downregulated TLR4 and MyD88 expressions in the liver. CONCLUSIONS CCA mitigates Sj-induced liver fibrosis by targeting TNF, TP53, JUN, and MMP9 to modulate the TLR4/MyD88 pathway, thereby suppressing pro-inflammatory cytokine release, inhibiting hepatic stellate cell activation, reducing collagen deposition, and preventing granuloma formation in the liver.
Animals ; Toll-Like Receptor 4/metabolism* ; Mice ; Myeloid Differentiation Factor 88/metabolism* ; Schistosoma japonicum ; Liver Cirrhosis/parasitology* ; Schistosomiasis japonica ; Signal Transduction ; Molecular Docking Simulation ; Inflammation ; Centella/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Tumor Necrosis Factor-alpha/metabolism*

Objective:To establish the HPLC fingerprint of Centellae herba and determine the content of asiaticoside, madecassic acid and asiaticoside B simultaneously; To compare the quality differences of Centellae herba collected in different months. Methods:The chromatographic condition was a Shimadzu InertSustain C18 column (4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile and 2 mmol/L beta cyclodextrin in gradient elution at the flow rate of 0.8 ml/min. The detection wavelength was 204 nm, and the column temperature was 30 ℃. The different Centellae herba materials of collected in 2-12 months from Chenzhou were studied by the similarity evaluation combined with cluster analysis, principal component analysis and the three contents determination. Results:The HPLC fingerprint of Centellae herba was established and 9 common peaks were designated. The eleven samples were different, which can be aggregated into 4 categories and the quality of Centellae herba collected in July was the best. Conclusion:The established fingerprint and multi-components quantitative method are stable and reliable, which can provide a reference for the quality control and the utilization of Centellae herba resource.

The traditional primary culture methods of human normal epithelial cells have disadvantages of low activity of cultured cells,the low cultivated rate and complicated operation.To solve these problems,researchers made many studies on culture process of human normal primary epithelial cell.In this paper,we mainly introduce some methods used in separation and purification of human normal epithelial cells,such as tissue separation method,enzyme digestion separation method,mechanical brushing method,red blood cell lysis method,percoll layered medium density gradient separation method.We also review some methods used in the culture and subculture,including serum-free medium combined with low mass fraction serum culture method,mouse tail collagen coating method,and glass culture bottle combined with plastic culture dish culture method.The biological characteristics of human normal epithelial cells,the methods of immunocytochemical staining,trypan blue exclusion are described.Moreover,the factors affecting the aseptic operation,the conditions of the extracellular environment,the conditions of the extracellular environment during culture,the number of differential adhesion,and the selection and dosage of additives are summarized.

Objective To study the effects of ischemic postconditioning and ischemic preconditioning on cerebral ischemia-reperfusion injury following middle cerebral artery occlusion in rats.Methods A reversible focal cerebral ischemia-reperfusion injury was modeled using middle cerebral artery occlusion. Thirty male Sprague-Dawley rats were randomized into three groups (n = 10 in each group) : a cerebral ischemia-reperfusion group, an ischemic postconditioning group and an ischemic preconditioning group. The impairment of neurological function was scored and the infarct volume, the activity of superoxide dismutase and malondiadehyde (MDA) content were measured after the operation.Results In the ischemic postconditioning and preconditioning groups the neurological function was better and the infarction volume was significantly smaller compared with the model group. In the preconditioning group both infarction volume and neurological function were significantly better than in the postconditioning group. In the brain tissues of the preconditioning and postconditioning groups MDA content was lower, while the activity of superoxide dismutase was significantly higher than in the model group.Conclusions lschemic postconditioning can attenuate pathological injury induced by focal cerebral ischemia and repedusion. The neuroprotective effect induced by ischemic preconditioning is stronger than that induced by ischemic postconditioning.

AIM:To observe the therapeutic effect of dihydromyricetin on experimental schistosoma japonicum hepatic fibrosis in mice. METHODS:60 mice infected with schistosma japonicum cercariae percutanoeusly were divided into 3 groups:model group,praziquantel group, praziquantel plus dihydromyricetin group and other 20 normal mice were used as control group.After treatment with medicine for 8 weeks,the liver was removed and weighed.The contents of ALT and AST in serum were assayed using the corresponding kits.Moreover, the degree of hepatic fibrosis was observed Via HE and was scored.The expression of collagenⅠprotein and collagenⅢprotein were measured by immunohistochernical method.RESULTS: The mice that infected with schistosoma japonicum, had a featuring increment in liver weights,serum ALT,AST contents,the expression of collagenⅠprotein,collagenⅢprotein (P