Research progress on the development of human neutrophil elastase inhibitors / 药学学报

Human neutrophil elastase (hNE) is a serine proteolytic enzyme mainly distributed in neutrophils. When the balance between anti-hNE protein and hNE is broken, excessive release of hNE can cause the occurrence of various diseases. Therefore, inhibition of hNE is a promising therapeutic strategy. In this paper, the structure, action mechanism, physiological function of hNE and the development of hNE inhibitors were briefly summarized, in order to provide information for the related research.

Research Progress of Human Neutrophil Elastase Inhibitors and Their Application in Diverse Diseases / 中国药学杂志

To comprehensively understand the research status of human neutrophil elastase inhibitors (HNEI) and their clinical application prospect, the papers about novel HNEI discovered since 2011 and diseases related to human neutrophil elastase (HNE) in addition to pulmonary diseases were summarized. The results showed that a lot of highly selective and potent HNE inhibitors have been discovered since 2011. HNE participates in the development of many diseases. In addition to the infectious and inflammatory pulmonary diseases reported in the past, it is also associated with ischemia-reperfusion injury, rheumatoid arthritis, autoimmune diabetes, nephritis, cancer and other diseases. The development of novel HNEI with high potency and low toxicity has been an important direction for HNE-related diseases.

Effects of Scutellarin on MUC5AC Mucin Production Induced by Human Neutrophil Elastase or Interleukin 13 on Airway Epithelial Cells

Scutellarin is a flavonoid extracted from a traditional Chinese herb, Erigeron breviscapus. The present study investigated the effect of scutellarin on MUC5AC mucin production and the possible mechanism. Human bronchial epithelial 16 (HBE16) cells were pretreated with scutellarin for 60 min, and then exposed to human neutrophil elastase (HNE) or interleukin (IL)-13 for 12 hr. RT-PCR and ELISA were performed to measure the amount of MUC5AC mucin production. The results showed that scutellarin inhibited MUC5AC expression both in mRNA and protein level induced by HNE in a concentration-dependent manner. However, scutellarin failed to inhibit MUC5AC mucin production induced by IL-13. To investigate the intracellular mechanisms associated with the effect of scutellarin on MUC5AC mucin production, western blotting was carried out to examine the phosphorylation of protein kinase C (PKC), signal transducer and activator of transcription 6 (STAT6) and extracellular signal-regulated kinase 1/2 (ERK1/2). The phosphorylation of PKC and ERK1/2 was attenuated after treatment with scutellarin, whereas STAT6 was not significantly affected. Therefore, it is suggested that scutellarin down-regulates MUC5AC mucin production on HBE16 cells via ERK-dependent and PKC-dependent pathways.

Roles of theaflavins in inflammatory airway mucus hypersecretion / 上海第二医科大学学报

Objective To investigate the roles of theaflavins(TFs) in airway mucus hypersecretion induced by human neutrophil elastase(HNE). Methods Human lung adenocarcinoma cell A549 was stimulated by HNE for mucus hypersecretion,and TF monomers(TF1,TF2 and TF3) were used for intervention.The effects of TF monomers on viability of A549 cells were examined by MTT method.After the effective doses of TFs were determined,A549 cells were divided into 4 groups for experiment.In control group,A549 cells were cultured with serum-free medium.In HNE treatment group,A549 cells were treated with HNE(50 nmol/L) for 24 h.In TF monomer intervention groups,A549 cells were pre-treated with TF1,TF2 or TF3(50,100 or 200 ?g/mL) for 24 h,and were then treated with HNE for another 24 h.In AG1478 intervention group,A549 cells were pre-treated with AG1478(5 ?mol/L),an epidermal growth factor receptor blocker for 30 min,and were then treated with HNE for another 24 h.The changes in mucin(MUC) after treatment by different doses of TF1,TF2 and TF3,and by TF3(200 ?g/mL) for different time(12 h,24 h and 36 h) were detected.The changes in MUC5AC mRNA expression and MUC5AC protein expression were detected by RT-PCR and ELISA,respectively. Results The MUC5AC mRNA expression and protein expression in HNE treatment group were significantly higher than those in control group(P

The Relationship between Human Neutrophil Elastase and Coronary Arterial Dilatation in Kawasaki Disease

PURPOSE: Kawasaki disease is notorious for coronary arterial complication which is usually developed as a febrile disease in early childhood. Increased polymorphonucleus(PMN) cell levels in acute phases may be associated with the pathophysiology of Kawasaki disease. We studied the relationship between coronary arterial dilatation and elastase activity which was excreted from PMN cell and roles as an important factor for vasculitis. METHODS: Ten patients diagnosed with Kawasaki disease in Yonsei University Medical Center were examined between November, 2001 and January, 2002. In addition, 15 patients with other febrile diseases were also examined. Echocardiography was done in patients with Kawasaki disease on the first day of admission and four weeks after the onset of the disease. At each time, venous samples were drawn and separated into plasma and leukocytes. In patients with other febrile disease, samples were drawn on admission. Elastase activities in plasma and neutrophil extracts were measured. RESULTS: The significant increased plasma elastase activity, 6.19+/-0.74 U/mL, found in Kawasaki disease patients compared with the other febrile disease patients, 4.86+/-1.17 U/mL(P<0.05). And there was no significance between the above two diseases in terms of the elastase activity in neutrophil extracts. The relationship between initial elastase activity and the coronary arterial complication which was shown in subacute phase wasn't significant. CONCLUSION: Plasma elastase activity was increased in Kawasaki disease significantly, but the initial plasma elastase activity in the acute phase could not reflect the range of coronary arterial complication.

Molecular characteristics of the inhibition of human neutrophil elastase by nonsteroidal antiinflammatory drugs

Nonsteroidal antiinflammatory drugs(NSAIDs) are known as clinically effective agents for treatment of inflammatory diseases. Inhibition of cyclooxygenase has been thought to be a major facet of the pharmacological mechanism of NSAIDs. However, it is difficult to ascribe the antiinflammatory effects of NSAIDs solely to the inhibition of prostaglandin synthesis. Human neutrophil elastase (HNElastase; HNE, EC 3.4.21.37) has been known as a causative factor in inflammatory diseases. To investigate the specific relationship between HNElastase inhibition and specificity of molecular structure of several NSAIDs, HNElastase was purified by Ultrogel AcA54 gel filtration, CM-Sephadex ion exchange, and HPLC (with TSK 250 column) chromatography. HNElastase was inhibited by aspirin and salicylate in a competitive manner and by naproxen, ketoprofen, phenylbutazone, and oxyphenbutazone in a partial competative manner, but not by ibuprofen and tolmetin. HNElastase-phenylbutazone-complex showed strong Raman shifts at 200, 440, 1124, 1194, 1384, 1506, and 1768 cm(-1). The Raman bands 1194, 1384, and 1768 cm(-1) may represent evidences of the conformational change at -N=N-phi radical, pyrazol ring, and -C=O radical of the elastase-drug complex, respectively. Phenylbutazone might be bound to HNElastase by ionic and hydrophobic interaction, and masked the active site. Inhibition of HNElastase could be another mechanism of action of NSAIDs besides cyclooxygenase inhibition in the treatment of inflammatory diseases. Different inhibition characteristics of HNE-lastase by NSAIDs such as aspirin, phenylbutazone-like drugs and ineffective drugs could be important points for drawing the criteria for appropriate drugs in clinical application.

Solid-phase enzyme. linked immunosorbent assay and its clinical significance for human neutrophil elastase / 中国病理生理杂志

The role of neutrophil in the pathogenesis of many diseases attracts more attentions than before. Neutrophil elastase (NE), a powerful neutrol proteinase capable of causing major tissue destruction in many diseases, is mainly present in the azurophil granules of neutrophil. A solid-phase enzyme-linked immunosorbent-biotin assay for human NE has been developed. Only 20?l of plasma sample is needed. Non-smoking healthy adults have 33.7?6.7ng NE/ml plasma; patients with chronic obstructive pulmonary diseases, 48.0+ 10.9 ng NE/ml; patients with pyogenic dermatisis, 69.1?15.7 ng NE/ml; patients with systemic lupus erythematosus, 61.0?13.9 ng NE/ml; patients with aplastic anemia, only 12.3?5.3 ng NE/ml. All of these results have significant differences compared with the result of healthy adults. The results of assay showed that this quantitative assay with good specificity and accuracy may be a better criterion for investigating the effect of NE on pathogenesis of some diseases.