I. Kazimierová

Echinacea complex--chemical view and anti-asthmatic profile.

ETHNOPHARMACOLOGICAL RELEVANCE Echinacea purpurea (L.) Moench is one of the mostly used herbs in the traditional medicine for the treatment of respiratory diseases. Modern interest in Echinacea is directed to its immunomodulatory activity. Recent studies have shown that secretion of asthma-related cytokines in the bronchial epithelial cells can be reversed by Echinacea preparations. AIM OF THE STUDY To examine the pharmacodynamics profile of Echinacea active principles, a complex has been isolated from its flowers by alkaline extraction and has been tested using an animal model of allergic asthma. MATERIAL AND METHODS The structural features of Echinacea purpurea complex was determined using chemical and spectroscopic methods. Allergic inflammation of the airways was induced by repetitive exposure of guinea pigs to ovalbumin. Echinacea complex was then administered 14 days in 50mg/kg b.w. daily dose perorally. Bronchodilatory effect was verified as decrease in the specific airway resistance (sRaw) in vivo and by reduced contraction amplitude (mN) of tracheal and pulmonary smooth muscle to cumulative concentrations of acetylcholine and histamine in vitro. The impact on mucociliary clearance evaluated measurement of ciliary beat frequency (CBF) in vitro using LabVIEW™ Software. Anti-inflammatory effect of Echinacea complex was verified by changes in exhaled NO levels and by Bio-Plex® assay of Th2 cytokine concentrations (IL-4, IL-5, IL-13 and TNF-alpha) in serum and bronchoalveolar lavage fluid (BALF). RESULTS Chemical and spectroscopic studies confirmed the presence of carbohydrates, phenolic compounds and proteins, as well as the dominance of rhamnogalacturonan and arabinogalactan moieties in Echinacea complex. The significant decrease in sRaw values and suppressed histamine and acetylcholine-induced contractile amplitude of isolated airways smooth muscle that were similar to effects of control drug salbutamol confirmed Echinacea complex bronchodilatory activity. The anti-inflammatory effect was comparable with that of control agent budesonide and was verified as significantly reduced exhaled NO levels and concentration of Th2 cytokines in serum and BALF. The values of CBF were changed only insignificantly on long-term administration of Echinacea complex suggested its minimal negative impact on mucociliary clearance. CONCLUSION Pharmacodynamic studies have confirmed significant bronchodilatory and anti-inflammatory effects of Echinacea complex that was similar to effects of classic synthetic drugs. Thus, results provide a scientific basis for the application of this herb in traditional medicine as a supplementary treatment of allergic disorders of the airways, such as asthma.

Bronchodilatory, antitussive and anti-inflammatory effect of morin in the setting of experimentally induced allergic asthma.

Using an experimental model of allergic asthma, we evaluated the anti‐asthmatic potential of polyphenol flavonol derivate morin after either acute or long‐term treatment of male OVA‐sensitised guinea pigs.

Potassium Ion Channels and Allergic Asthma

High-conductive calcium-sensitive potassium channels (BK+Ca) and ATP-sensitive potassium (K+ATP) channels play a significant role in the airway smooth muscle cell and goblet cell function, and cytokine production. The present study evaluated the therapeutic potential of BK+Ca and K+ATP openers, NS 1619 and pinacidil, respectively, in an experimental model of allergic inflammation. Airway allergic inflammation was induced with ovalbumine in guinea pigs during 21 days, which was followed by a 14-day treatment with BK+Ca and K+ATP openers. The outcome measures were airway smooth muscle cells reactivity in vivo and in vitro, cilia beating frequency and the level of exhaled NO (ENO), and the level of pro-inflammatory cytokines in the plasma and bronchoalveolar lavage fluid. The openers of both channels decreased airway smooth muscle cells reactivity, cilia beating frequency, and cytokine levels in the serum. Furthermore, NS1619 reduced ENO and inflammatory cells infiltration. The findings confirmed the presence of beneficial effects of BK+Ca and K+ATP openers on airway defence mechanisms. Although both openers dampened pro-inflammatory cytokines and mast cells infiltration, an evident anti-inflammatory effect was provided only by NS1619. Therefore, we conclude that particularly BK+Ca channels represent a promising new drug target in treatment of airways allergic inflammation.

The effect of short-term and long-term application of fisetin on experimentally induced airway hyperreactivity

Abstract Background: Fisetin, a derivate from the flavonol group may possess a variety of pharmacological effects. The aim of the presented study was to evaluate the bronchodilatory effect of fisetin after the acute or the chronic administration to guinea pigs with allergic airway inflammation. Methods: Experimental animals were sensitized and challenged by ovalbumin. Fisetin was administered in dose 5mg/kg/p.o., either once after the end of 21-days sensitization or daily during the 21-days sensitization. By using the whole-body plethysmograph, we monitored the specific airway resistance, a parameter of airway hyperreactivity in vivo. The changes of the specific airway resistance were evaluated after the short-term inhalation of the bronchoconstriction mediator-histamine (10−6 mol.1−1). Results: Our results showed that the short-term as well as the long-term administration of fisetin caused decrease of the specific airway resistance values. The bronchodilatory effect of fisetin was comparable to the long-acting beta2 sympathomimetic – salmeterol after the long-term administration. The measurements of the bronchodilatory activity after single administration have revealed more prolonged effect of fisetin comparing to the short-acting beta2 sympathomimetic – salbutamol, as this remained even after the 5 hours, when salbutamol was already ineffective. Conclusion: In conclusion, flavonol – fisetin has shown bronchodilatory potential. In the light of this fact, fisetin may represent potential substance that can be effective in both prevention as well as control of airway inflammation symptoms.

The role of cytokines in degenerative spine disorders

Abstract Background: Degenerative spine disorders (DSD) are the most frequent reason of morbidity in adults. Commonly DSD includes degenerative disorders of intervertebral discs (IVDs), spinal stenosis and degenerative spondylolisthesis (SL). There is increasing evidence about significant role of cytokines in DSD pathogenesis, symptomathology and progression, but their protective levels remain still unknown. Material and Methods: The aim of presented study was to provide quantitative and qualitative analysis of cytokine, chemokine and growth factors levels in individual parts of IVDs - annulus fibrosus (AF) and nucleus pulposus (NP) - separately and in facet joints (FJ) subchondral bone of patients with DSD and in controls - healthy subjects during a multiorgan procurement procedure. Bio-Plex® assay was used to measure concentrations of 27 different cytokines in tissue of patients with DSD. Their concentrations in tissues of healthy subjects during a multiorgan procurement procedure represented protective levels. Results: The Bio-Plex® assay revealed significant differences between the patients suffered from degenerated and herniated IVDs and from lumbar SL and controls in cytokines, chemokines and growth factor profiles suggested that pro-inflammatory changes of both NP and AF were dominated in herniated IVDs, whereas the same tissue of lumbar SL patients exhibited much more complex changes in cytokine levels suggested o only ongoing inflammation (IL-6, IL-8, MCP-1, TNF-α), abut also antiinflammatory processes (IL-ra, IL-10) or connective tissue remodeling (PDGF-bb, IL-17, VEGF). The different mediators were found elevated in lumbar SL samples of subchondral FJ bone. These also confirmed ongoing inflammation, accelerated bone resorption and formation and increased fibroblasts activity in FJ bone. Conclusion: The study supported the significant involvement of several cytokines, chemokines and growth factors in the pathogenesis of DSD. These cytokines should represent future potential targets for new biological treatment able to slow DSD progression as well as factor determining prognosis of DSD.

Chemico-physical and pharmacodynamic properties of extracellular Dictyosphaerium chlorelloides biopolymer

Microalgae occupy all territories and their products represent a rich source of phytochemicals for human being. Green microalga Dictyosphaerium chlorelloides was found to be a significant producer of the extracellular biopolymer. Dominant components of the biopolymer were found to be Gal (39 wt%) with its methyl derivatives (15 wt%), Rha (19 wt%) and Man (14 wt%). 2-OMe-Gal was found to be the major derivative while other sugars, namely 3-OMe-, 6-OMe- and 2,3-di-OMe-Gal, 3-OMe-Glc and 4-OMe-Xyl were in smaller amounts. NMR spectroscopy revealed complex structure with galactan backbone branched by sugars in furano and pyrano forms in alpha and beta configurations. NMR data of 2-OMe, 3-OMe, 2,3-OMe and 6-OMe galactoses afforded characteristic values for O-methyls in each position. Biopolymer antitussive effect was similar to that of centrally acting antitussive drugs, indicating its relatively good antitussive potential.

Acute and Chronic Effects of Oral Erdosteine on Ciliary Beat Frequency, Cough Sensitivity and Airway Reactivity

Erdosteine as a mucolytic agent that decreases mucus viscosity and facilitates mucus expulsion from the airways by cough or ciliary movement. Our objective was to determine whether erdosteine can directly contribute to mucus clearance. We addressed the issue by monitoring acute and chronic effects of erdosteine on ciliary beat frequency (CBF), cough sensitivity, and airway smooth muscle reactivity. The experiments were performed in healthy guinea pigs. Erdosteine (10 mg/kg) was administrated orally in a single dose or daily through 7 days. The cough reflex and specific airway resistance were evaluated in vivo. The CBF in tracheal brushed samples and the contractile response of tracheal smooth muscle stripes to bronchoconstrictive mediators were evaluated in vitro. We found that neither acute nor chronic erdosteine treatment had a significant effect on cough sensitivity and airway reactivity. However, in the vitro condition, erdosteine increased CBF and reduced tracheal smooth muscle contractility; the effects were more pronounced after chronic treatment. We conclude that erdosteine may directly contribute to mucus clearance by CBF stimulation. Although erdosteine has no effect on cough reflex sensitivity, its mild bronchodilator and mucolytic properties may promote effective cough.

Influence of smooth muscle contractility by inhibition of crac channels activity

Abstract The present in vitro study was focused on the differences in expression and activity of calcium release-activated calcium (CRAC) channels of human term-pregnant and non-pregnant myometrium. The expression of Orai1 protein, as a functional subunit of CRAC channel, was significantly higher than in non-pregnant myometrium. Lower Orai1 protein expression did not influence the amplitude of contractile response of term-pregnant myometrium, but higher Orai1 expression observed in non-pregnant myometrium was related to the different influence of CRAC blocker on contraction frequency.

Effect of inhaled and oral n-acetylcysteine on airway defense mechanism

Abstract Aim: N-acetylcysteine is the prototype of mucolytic agents. The aim of this study was to evaluate the acute and chronic effect of inhaled and oral N-acetylcysteine on airway reactivity, cough reflex and ciliary beat frequency and parameters of mentioned defense mechanisms were assessed in physiological conditions. Methods: An experiment was performed using healthy guinea pigs treated with inhaled (0.6 M; 5min) and oral N-acetylcysteine (20 mg/kg), administrated either acutely as a single dose or chronically during 7 days. The cough reflex and specific airway resistance were assessed by in vivo method, using a double chamber plethysmograph box. The ciliary beat frequency was evaluated in in vitro conditions on tracheal brushed samples using light microscope coupled to high speed video camera. Results: Inhaled and oral N-acetylcysteine, either administrated as a single dose or during 7 days, have shown a tendency to decrease sensitivity of the cough reflex and increase the airway reactivity. Acute administration of inhaled and oral N-acetylcysteine had no statistically relevant effect on the ciliary beat frequency, whereas chronic administration of both inhaled and oral N-acetylcysteine led to a marked reduction in the ciliary beat frequency. Conclusion: Chronic administration of oral and inhaled N-acetylcysteine had a negative impact on the ciliary beat frequency, which represents one of the key factors determining the rate of mucociliary clearance. Thus, administration of N-acetylcysteine is less likely to increase the expulsion of mucus by ciliary movement. In addition, the observed tendency of inhaled and oral N-acetylcysteine to increase the airway reactivity may limit its use in conditions with severe airflow obstruction.