Mechanism of Chaenomelis Fructus in treatment of rheumatoid arthritis based on network pharmacology and experimental verification / 中国中药杂志

The material basis and mechanism of Chaenomelis Fructus in the treatment of rheumatoid arthritis(RA) were explored by network pharmacology, and the potential anti-RA targets of Chaenomelis Fructus were verified by molecular docking and animal experiments. The active components and targets of Chaenomelis Fructus were searched against the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform. GeneCards, DisGeNET, and OMIM were used to obtain RA-related targets. The common targets shared by Chaenomelis Fructus and RA were considered as the potential targets of Chaenomelis Fructus in the treatment of RA. Cytoscape 3.9.0 was employed to establish a "traditional Chinese medicine-active component-common target-disease" network. The protein-protein interaction(PPI) network was established by STRING, and the core genes were visualized by RStudio 4.1.0. DAVID was used for Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment to predict and visualize the involved signaling pathways. Molecular docking was carried out with the active components screened out as ligands and RA core genes as the targets. Finally, the prediction results were verified by animal experiments. Four main active components of Chaenomelis Fructus were obtained, which corresponded to 137 targets. Chaenomelis Fructus and RA shared 37 common targets. GO annotation yielded 239 terms(P<0.05), and KEGG pathway enrichment analysis screened out 94 signaling pathways(P<0.05), mainly involving interleukin-17(IL-17), tumor necrosis factor, Toll-like receptor, and nuclear factor-kappa B(NF-κB) signaling pathways. Molecular docking results showed that the main active components of Chaenomelis Fructus bound well with the core targets of RA. The results of animal experiments proved that Chaenomelis Fructus can alleviate joint swelling in the mice with RA. The results of ELISA showed that Chaenomelis Fructus lowered the levels of interleukin-6(IL-6) and interleukin-1β(IL-1β). Western blot showed that Chaenomelis Fructus down-regulated the protein level of vascular endothelial growth factor A(VEGFA). Chaenomelis Fructus exerts anti-inflammatory effect and reduces pannus formation by regulating the core targets such as VEGFA, IL-1β, and IL6 in the treatment of RA. The findings of this study provide new ideas for the future treatment of RA with Chaenomelis Fructus.

Analysis of related factors of neoatherosclerosis after coronary stent implantation by optical coherence tomography / 中国介入心脏病学杂志

Objective To evaluate the risk factors of neoatherosclerosis after coronary stent implantation through the study of optical coherence tomography (OCT) observation, and the effect of statin lipid-lowering therapy. Methods The clinical data of 40 patients with coronary heart disease (CHD) after coronary stent implantation (OCT) in Peking University First Hospital from June 2013 to June 2016 were analyzed retrospectively. The patients were divided into the target-achieved group (19 cases, LDL-C<1.8 mmol/L) and the above-target group (21 cases LDL-C ≥ 1.8 mmol/L) according to whether their low density lipoprotein (LDL) levels reached the treatment target. The medical history, biochemical parameters, medication and the qualitative and quantitative analysis of the neoatherosclerosis in the two groups were reviewed and compared. Results There were no statistical differences between the two groups in age, sex, smoking, comorbidities of hypertension, diabetes mellitus, kidney disease, history of myocardial infarction and time of stent implantation. Patients in the above-target group had higher LDL-C levels[(2.42 ± 0.79)mmol/L vs.(1.30 ± 0.35)mmol/L,P<0.001],higher cholesterol levels[(4.38 ±1.01)mmol/L vs.(3.07± 0.87)mmol/L,P<0.001], but lower statin used dosage[(15.26 ±2.30)mg/d vs.(22.38 ± 2.10)mg/d,P=0.032]as compared to patients in the target-achieved group. There was no statistical difference between the other parameter and medication profile. Qualitative analysis showed higher rates of neoatherosclerosis in the above-target group as compared to the target-achieved group(78.9% vs.38.1%,P=0.009),plus higher rates of endothelial heterogeneity(78.9% vs.38.1%,P=0.009),lipid plaque(73.7% vs.38.1%,P=0.024) and calcified plaque(26.3% vs. 4.8%,P=0.049). The rate of neointima restenosis in the above-target group was higher than that of the target-achieved group[(25.12 ±1.10)% vs.(19.70±1.10)%, P=0.001]. Logistic analysis showed that the level of low density lipoprotein(P=0.009),dosage of statin prescribed (P=0.040), male gender (P=0.042) and serum creatinine (P=0.012) were associated with the present of neonatal arteriosclerosis .Multivariate regression analysis showed levels of lipoprotein and low presence of statin dosage were the independent wish factors. Low density lipoprotein levels were positively correlated with neoatherosclerosis formation and the dosage of statin was negatively correlated (P<0.05).Conclusions In-stent restenosis after PCI is closely related to neonatal atherosclerosis. Controlling LDL-C to target levels can significantly reduce the risk of neoatherosclerosis. Intensified statin therapy strategy can prevent neoatherosclerosis in by certain extent.

Effects of pargyline on histone methylation in promoter and enhancer regions and transcription of CYP3A4/3A7 / 药学学报

This study was designed to investigate effects of pargyline on histone methylation in the promoter and enhancer regions and transcription of cytochrome P450 3A4/3A7 (CYP3A4/3A7) gene. Human primary fetal liver cells were isolated, cultured and randomly divided into several groups including control, solvent, pargyline low, middle, high dose (treated with 0.6, 1.2, 2.4 mmol·L-1). HepG2 cells were cultured and treated with 0.03, 0.3, 3 mmol·L-1 pargyline. After 48 hours, total RNAs were prepared from the cells to determine the expression of CYP3A mRNA in primary fetal cells and HepG2 cells with real-time quantative PCR (qPCR). HepG2 cells were cultured and then treated with 3 mmol·L-1 pargyline for 48 hours. The chromatin immunoprecipitation (ChIP) assay was performed with dimethylation of histone H3 at lysine 4 (H3K4me2), and IgG antibodies respectively. The precipitated DNA was resuspended and used for qPCR. Primers were used to detect different regions of CYP3A4/3A7 promoter and enhancer. Occupancy of H3K4me2 was shown as percent of input DNA relative to control cells. The results suggested that pargyline has an effect on primary fetal liver cells and HepG2 cells proliferation. The level of CYP3A7 was markedly enhanced in human primary fetal liver cells by treatment with 1.2, 2.4 mmol·L-1 of pargyline (P-1 of pargyline in HepG2 cells (P<0.001) compared with solvent control. Occupancy of H3K4me2 on human CYP3A4 promoter (-362 to +53) and enhancer segment (-7 836 to -6 093) harbored the overlapping hepatocyte nuclear factors 4A (HNF4A) binding site compared with a negative control. Occupancy of H3K4me2 on human CYP3A7 promoter (-163 to +103) and enhancer segment (-4 054 to -3 421, -6 265 to -6 247) overlapped with glucocorticoid receptor (GR) binding site. In conclusion, the enriched H3K4me2 in the promoter and enhancer regions was induced by pargyline with HNF4A or GR binding site in CYP3A4/3A7 gene to activate the corresponding genes.