ABSTRACT
To explore the role of hydrogen peroxide (H2O2) in promoting polymorphonuclear neutrophils adherence and injury of human umbilical vein endothelial cells (HUVECs), the ordinary optical microscope and scanning electron microscopy were used to observe the adherence and injury after HUVECs co-cultured with neutrophils pretreated by extracellular H2O2 (HUVECs and neutrophils co-culture without H2O2 pretreatment as control), and the adhesion rates of neutrophils were measured through cell count test. The percentages of HUVECs expressing intercellular adhesion molecule 1 (ICAM-1) and Apo2.7 were detected by flow cytometry. After being cocultured with the neutrophils pretreated by extracellular H2O2, HUVECs showed obvious injury changes, such as round or oval shape, shortened or disappeared microvilli, and membrane structural damage; The adhesion rate of neutrophils was (57.74 ± 9.18)%, which was significantly higher than that in control [(23.12 ± 6.43)%, P < 0.01, n = 8]; The percentages of HUVECs expressing ICAM-1 and Apo2.7 were (44.69 ± 1.52)% and (39.29 ± 1.81)% respectively, which were significantly higher than those in control [(21.79 ± 1.43)% and (9.79 ± 1.43)%] (P < 0.01, n = 8). The results suggest that extracellular H2O2 can promote the neutrophils adherence and injury of HUVECs.
Subject(s)
Humans , APOBEC Deaminases , Cell Adhesion , Coculture Techniques , Cytidine Deaminase , Metabolism , Human Umbilical Vein Endothelial Cells , Cell Biology , Hydrogen Peroxide , Pharmacology , Intercellular Adhesion Molecule-1 , Metabolism , Muscle Proteins , Metabolism , Neutrophils , Cell BiologyABSTRACT
Isocitrate lyase (ICL) which plays a pivotal role in the glyoxylate cycle of mycobacterium tuberculosis(MTB) could be a potential target against mycobacterium tuberculosis.Deoxyribozyme(DRz) is a small single-strand,enzymatic DNA molecule.Assistant by sketch of secondary structure,appropriate and not appropriate target sequences of MTB ICL mRNA were predicted.Two DRzs were designed targeting above sequences respectively.To affirm feasibility of this kind of prediction,in vitro cleavage of MTB ICL mRNA segment by DRz-ICLcf and DRz-ICLcj were performed.The results showed that sketch of secondary structure could indeed predict potential target of DRz.This research will provide basis for developing DRz as a novel antimicrobial agent against MTB.
ABSTRACT
Human cytomegalovirus (HCMV) is a DNA virus and serious opportunistic pathogen for both newborn and immunocompromised individuals.To research technique for gene silence and antiviral agents, ribozyme M1GS-T6 was constructed from external guide sequences(EGSs)that consist of a sequence complementary to HCMV UL54 gene RNA and M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli. The results showed that M1GS can efficiently cleave the mRNA sequence encoding UL54 protein in vitro.