Inhaled Cissampelos sympodialis Down-Regulates Airway Allergic Reaction by Reducing Lung CD3+T Cells.

Phytother Res. 2013 Jun;27(6):916-25.

Vieira GC, De Lima JF, De Figueiredo RC, Mascarenhas SR, Bezerra-Santos CR, Piuvezam MR.

Laboratory of Immunopharmacology, Department of Physiology and Pathology, Federal University of Paraíba, João Pessoa, Paraíba, Brazil.

ABSTRACT:

Background: Cissampelos sympodialis Eichl. (Menispermaceae) root infusion is used in Northeast Brazil to treat allergic asthma. We have previously shown that oral use of the plant extract reduces eosinophil infiltration into the lung of ovalbumin (OVA)- sensitized mice. However, drugs taken by inhalation route to treat asthma achieve better
outcomes.

Objetive: Thereby, in this study, we evaluated the inhaled C. sympodialis leaves hydroalcoholic extract as a therapeutic treatment in OVA-sensitized BALB/c mice.

Methods: The parameters analyzed consisted of leukocyte recruitment to the airway cavity, tissue remodeling and cell profile.

Results: The inhaled extract inhibited mainly eosinophil recruitment to the pleural cavity, bronchoalveolar lavage and peripheral blood. This treatment reduced the OVA-specific IgE serum titer and leukocyte infiltration in the peribronchiolar and pulmonary perivascular areas as well as mucus production. In addition, we also tested isolated alkaloids from the plant extract. The flow cytometric analysis showed that methylwarifteine (MW) and, mainly, the inhaled extract reduced the number of CD3+T cells and eosinophil-like cells.

Conclusion: Therefore, inhaled C. sympodialis extract and MW lead to downregulation
of inflammatory cell infiltration with remarkable decrease in the number of T cells in an experimental model of respiratory allergy. These data suggest that the plant can be delivered via inhalation route to treat allergic asthma.

PMID: 22933368

SUPPLEMENT:

Respiratory diseases such as asthma, bronchitis, rhinitis and chronic obstructive pulmonary disease are chronic inflammatory allergic illnesses characterized by variable airflow obstruction, airway hyperresponsiveness and tissue remodeling in association with airway inflammation.

The use of inhaled medications to treat respiratory diseases has been of great importance to medicine for decades, providing high drug concentration, rapid onset of action, low systemic exposure, thereby, reducing side effects. However, most drugs used to treat respiratory diseases as corticosteroids, b2-adrenergic agonists, phosphodiesterase (PDE) inhibitors and leukotriene receptor antagonists control the illness symptoms but also induce long-term side effects. Therefore, classical pharmacotherapy limitations to treat asthma and other allergic conditions open a research field to find alternative drugs to control these diseases.

The root infusion of Cissampelos sympodialis Eichl. (Menispermaceae) (Figure 1) is usually used in folk medicine in Northeast Brazil to treat inflammatory illnesses including asthma, bronchitis, colds and rheumatism. Previous studies using the hydroalcoholic fractions of roots or leaves (AFL) of C. sympodialis demonstrated immunomodulatory effect, suppressed the inflammatory process, inhibited the production of IgE and were associated with a decrease of IL-5 and IL-13 but also with increase of IFN-gamma. The bisbenzylisoquinoline alkaloids warifteine (WAF) inhibited eosinophil and neutrophil migration into the pleural cavity and bronchoalveolar lavage (BAL) of OVA-sensitized BALB/c mice, and it also reduced eotaxin levels, cytoplasm lipid body formation and cis-leukotriene generation in human eosinophils. WAF also inhibited spleen cell proliferation mediated by antigen or mitogen and reduced the antigen-specific-IgE-induced mast cell degranulation. Together, these data provide strong support for the proposition that the plant extract, as well as WAF, induces a modulatory effect on allergic and inflammatory reactions. Based on the available information on C. sympodialis, we studied the effect of the plant extract (AFL) and its alkaloids by inhalation as a therapeutic protocol in an experimental model of respiratory allergy.

Isogenic female BALB/c mice (7–8-week-old), weighing between 25 and 30 g, and female Wistar rats, weighing on an average 200 g, were used in the experimental protocols. Five groups (n=5–10) of female BALB/c mice were used in all experimental protocols, and they were divided in: saline group (non-sensitized animals), OVA group (OVA-sensitized animals and treated with saline), AFL 10 or AFL 100 groups (OVA-sensitized animals and treated with 10 or 100 mg/mL of AFL, respectively), Dexa group (OVA-sensitized animals and treated with 0.1 mg/mL of dexamethasone). For the histological analysis, a naive animal group was added to demonstrate influence of the inhalation procedure.

Inhaled AFL inhibited the inflammatory cell migration into the pleural cavity and into the BAL 24 h after OVA challenge. The doses of 10mg/mL or 100mg/mL induced a significant decrease in the total number of cells into the pleural cavity and into the BAL when compared to the OVA group. Influx of mononuclear cells into the pleural cavity and into the BAL was also significantly reduced in both AFL and dexamethasone treatments. Neutrophils were absent in the pleural cavity and significantly reduced into the BAL in AFL-treated animals. AFL or dexamethasone treatments also showed a significant reduction in the number of eosinophils into the pleural cavity as well as into the BAL.

The present study demonstrated that inhaled AFL (100 mg/mL) and the alkaloid isolated from the extract metilwarifteine (MW) efficiently reduced the CD3+T cell and eosinophil counts. Inhaled AFL at 100 mg/mL significantly decreased OVA-specific IgE serum titer in OVA-sensitized animals. Furthermore, our data demonstrated, for the first time that inhaled AFL has a systemic effect in modulating B cells.

For the histological analysis of six lung sections of each animal group: saline; OVA; AFL 10; AFL 100; naive animal and dexa; were stained with H&E, Gomori’s trichrome or PAS stains. The H&E staining allowed us to observe that 10 mg/mL of AFL did not inhibit cell migration. On the other hand, 100 mg/mL of AFL or dexa decreased cell infiltration into the peribronchiolar and perivascular regions. The Gommori’s Trichrome staining allowed us to observe that 100 mg/ml of AFL or dexamethasone were delivered and showed collagen fibers inhibition. However, 10 mg/mL of AFL did not present effects on this parameter. The PAS staining demonstrated that only 100 mg/ml of AFL inhibited the mucus production in the terminal and respiratory bronchioles, similar to what it was observed in the dexa group.

This study demonstrated that (Fig. 2) inhaled C. sympodialis treatment of animals with allergic airway inflammation is as effective as the oral treatment in controlling lung inflammatory response and the production of IgE. In addition, T cell immumodulatory effect of the isolated alkaloid MW and the C. sympodilais extract clearly provides the scientific evidence to support its medicinal efficacy in the management of allergic inflammatory diseases orchestrated by T cells. In addition, the results allowed us to suggest that the hydroalcoholic extract of the leaves of Cissampelos sympodialis can be the plant material for a phytotherapic medicine to be delivered by inhalation to prevent airway remodeling.

References:

Bezerra-Santos CR, Balestieri FMP, Rossi-Bergmann B, Peçanha LMT, Piuvezam MR. 2004. Cissampelos sympodialis Eichl. (Menispermaceae): oral treatment decreases IgE levels and induces a Th-1 shewed cytokine production in ovalbuminsensitized
mice. J Ethnopharmacol 95:1991–1997.

Bezerra-Santos CR, PeçanhaLMT, PiuvezamMR. 2005. Cissampelos sympodialis Eichl. (Menispermaceae) inhibits anaphylactic shock reaction in murine allergic model. Rev Bras Farmacogn 15:287–291.

Bezerra-Santos CR, Vieira-de-Abreu A, Barbosa-Filho JM, Bandeira-Melo C, Piuvezam MR, Bozza PT. 2006. Anti-allergic properties of Cissampelos sympodialis and its isolated alkaloid warifteine. Int Immunopharmacol 6:1152–1160.

Costa HF, Bezerra-Santos CR, Barbosa-Filho JM, Martins MA, Piuvezam MR. 2008. Warifteine, a bisbenzylisoquinoline alkaloid, decreases immediate allergic and thermal hyperalgesic reactions in sensitized animals. Int Immunopharmacol 8:519–525.

Piuvezam MR, Peçanha LMT, Alesander J, Thomas G. 1999. Cissampelos sympodialis Eichl Leaf extract increases the production of il-10 by concanavalin-A treated BALB/c spleen cells. J Ethnopharmacol 67: 93–101.

Contact:

Dr. Marcia Regina Piuvezam,
Department of Physiology and Pathology, Federal University of Paraíba, João Pessoa,
58051-970, Paraíba, Brazil.
E-mail: mrpiuvezam@ltf.ufpb.br

Fig 1. Cissampelos sympodialis Eichl. (Menispermaceae) Fig 2. Summary of results.

Inhaled Cissampelos sympodialis Down-Regulates Airway Allergic Reaction by Reducing Lung CD3+T Cells.

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