Elsevier

Clinica Chimica Acta

Volume 310, Issue 2, 20 August 2001, Pages 113-122
Clinica Chimica Acta

Serum concentration and circadian profiles of cathepsins B, H and L, and their inhibitors, stefins A and B, in asthma

https://doi.org/10.1016/S0009-8981(01)00530-7Get rights and content

Abstract

Background: In order to determine the effect of asthma on serum concentrations of cathepsins B, H and L, and stefins A and B, the circadian and concentration profiles were followed in steroid-independent and steroid-dependent asthmatics before and after 1-week treatment with methylprednisolone and cyclosporin A. Methods: Serum samples were taken at 4-h intervals throughout a 24-h period. Cathepsin and stefin concentrations were assayed using specific ELISAs. Data were analysed by one-way ANOVA and least squares fit of 24-h cosine. Results: Temporal analysis of these proteins revealed little or no significant changes with time over a 24-h period. In comparison to normal sera, cathepsin H concentrations were elevated in all asthmatic patients, concentrations of both stefins were decreased in steroid-independent asthmatics, and stefin A concentrations were increased in steroid-dependent asthmatics before therapy. The effect of methylprednisolone treatment was demonstrated on decreased cathepsin B and increased cathepsin L concentrations in post-therapy serum samples. On the other hand, cyclosporin A treatment led to increased concentrations of cathepsins H and L. However, concentrations of stefins A and B were unaffected. Conclusions: This study associated alterations in balance of serum cysteine proteinases and their inhibitors in asthmatic patients, which has raised the possibility of their involvement in asthma pathogenesis. Validated rhythms of cathepsins and stefins in asthmatic sera exhibited temporal differences, which are too small to influence the time of sampling for their quantitative measurement over the course of a day.

Introduction

Cysteine proteinases such as cathepsins B (EC 3.4.22.1), H (EC 3.4.22.16), and L (EC 3.4.22.15) constitute a major component of the lysosomal proteolytic system responsible for intracellular protein degradation and turnover [1]. They are also involved in specific cellular processes including prohormone activation and antigen processing [1], [2], [3]. Cysteine proteinases are capable of degrading components of the extracellular matrix, and have been implicated in normal and pathological processes of cell growth and tissue remodelling [2], [3], [4], [5]. By regulating cysteine proteinase activities, naturally occurring inhibitors, such as intracellular stefins and extracellular cystatins and kininogens, also play an important role in physiological and pathological processes [1], [6]. Altered concentrations or activities of cysteine proteases and their inhibitors have been implicated in serious human disorders such as cancer, autoimmune diseases, kidney failure, sepsis, arthritis, Alzheimer's disease, multiple sclerosis, epilepsy, psoriasis, osteoporosis, muscular dystrophy, inflammation, etc [1], [6]. It is suggested that their serum levels may be of clinical importance for prognosis and diagnosis, especially in cancer [7], [8], [9], [10], [11], [12].

Asthma is characterised by chronic airway inflammation with increased plasma exudation into the airway lumen, abnormal local secretion of proteins and airway wall remodelling. Asthmatic inflammation is still poorly defined and difficult to assess. Many inflammatory cells, mediators, and enzymes are involved, but their relative importance is not yet clear. Evidence is emerging that obstructive lung diseases such as asthma may result from a disturbed balance between proteases and their inhibitors [13]. In this report we have followed, over a 24-h period, concentration variations of cathepsins B, H and L, and their low molecular weight inhibitors stefins A and B, in sera from patients with steroid-independent and steroid-dependent asthma, before and after 1 week of treatment with methylprednisolone and cyclosporin A.

Section snippets

Subjects

The 21 volunteers who participated in the present study were recruited from the University Clinic of Respiratory and Allergic Diseases, Golnik. Written informed consent was obtained from all subjects. The procedures were approved and performed in accordance with the guidelines of the regional medical ethics committee. Meals were served at 08:00, 12:30 and 18:00 h. Patients and healthy subjects were not restricted in their water intake, except for the half-hour before sampling, but were asked to

Results

Table 1, Table 2, Table 3, Table 4, Table 5 show the mean 24-h concentrations, the differences between the maximum and the minimum concentrations over 24 h, and circadian characteristics of cathepsins B, H and L, and stefins A and B, which were determined by specific ELISAs in sera from steroid-independent asthmatics before (group B) and after (group C) treatment with methylprednisolone, and in steroid-dependent asthmatics before (group D) and after (group E) treatment with cyclosporin A. For

Discussion

Although cathepsins B, H and L, and stefins A and B are intracellular proteins, their presence at different concentrations has been observed in several body fluids including blood [1], [18]. Changed concentration profiles of cysteine proteinases and their inhibitors in blood may mirror the pathological processes where extensive proteolysis and tissue remodelling may occur, as has been already shown in cancer [7], [9], [10], [11], [12], [19], liver cirrhosis [19], pancreatitis [20], renal

Conclusion

According to the temporal analysis performed by ANOVA and cosinor analysis, the time of serum sampling in asthmatic patients does not compromise the results of quantitative measurements of cathepsins B, H and L, and stefins A and B over the course of a day, regardless of whether patients were subjected to methylprednisolone or cyclosporin A therapy, supporting their usefulness for clinical evaluation as diagnostic tools.

Together with our previous findings on cystatin C [17], our results provide

Acknowledgements

We thank Prof. Roger H. Pain for the critical reading of the manuscript. The work was supported in part by a grant from the Ministry of Science and Technology of Slovenia.

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