UPLC-ESI-QTOF-MS2 characterisation of Cola nitida resin fractions with inhibitory effects on NO and TNF-α released by LPS-activated J774 macrophage and on Trypanosoma cruzi and Leishmania amazonensis.

INTRODUCTION: The resin of Cola nitida is used in western Cameroon as incense for spiritual protection and during ritual ceremonies. This plant secretion has never been investigated although previous chemical and biological studies on other resins have drawn many attentions. OBJECTIVE: The resin fractions which revealed inhibitory effect on nitric oxide (NO) and tumour necrosis factor alpha (TNF-α) released by lipopolysaccharide (LPS)-activated J774 macrophage as well as on intracellular forms of Leishmania amazonensis and Trypanosoma cruzi amastigote were chemically characterised. Moreover, their antiparasitic activities were compared to those of semi-synthetic triterpenes. METHODOLOGY: The anti-inflammatory activity was evaluated by measuring the nitrite production and the TNF-α concentration in the supernatants of LPS-activated macrophages by antigen capture enzyme-linked immunosorbent assay. Moreover, the antiparasitic assay was performed by infecting the host cells (THP-1) in a ratio parasite/cell 10:1 (L. amazonensis) and 2:1 (T. cruzi) and then exposed to the samples. The resin was separated in vacuo by liquid chromatography because of its sticky behaviour and the chemical profiles of the obtained fractions (F1-F4) were established by dereplication based on UPLC-ESI-MS2 data while semi-synthetic triterpenes were prepared from α-amyrin by oxidation reactions. RESULTS: Fractions F1-F4 inhibited NO and TNF-α almost similarly. However, only F1, F3 and F4 showed promising antiparasitic activities while F2 was moderately active against both parasites. Hence, F1-F4 were exclusively composed of pentacyclic triterpenes bearing oleanane and ursane skeletons. Semi-synthetic compounds revealed no to moderate antiparasitic activity compared to the fractions. CONCLUSION: Although it will be difficult to prove the interaction resin-spirit, interesting bioactivities were found in the resin fractions.

Inhibitory effect of hydroalcoholic extract of Cunila spicata Benth. on phlogistic agents-induced cellular migration in the airways of mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Cunila spicata Benth. Is a creeping and aromatic plant that has an ethnopharmacological indication in the southern region of Brazil, in the Araucaria Forest biome. It's used as an alternative therapy for respiratory diseases for men and animals since the "tropeirista" movement in colonial Brazil. AIM OF THE STUDY: Investigate the influence of oral administration of hydroalcoholic extract of C. spicata (HECS) on cellular inflammatory processes in the airways of mice. MATERIAL AND METHODS: The HECS was obtained by the ultrasound-assisted technique. Phytochemical analysis of the volatile compounds was performed using gas-chromatography with mass spectrometry. To investigate the biological activity of HECS, the animals were pre-treated with HECS orally 1 h before and the bronchoalveolar lavage (BAL) model was used, in which the animals were subjected to inhalation of different pro-inflammatory agents such as carrageenan (CAR), histamine (HIST), capsaicin (CAP), bradykinin (BK), and 48/80 compound (C48/80). After sacrifice, BAL sample was collected from the bronchi and was analyzed for total and differential white blood cell counts, compared with control groups. RESULTS: Fourteen volatile phytocompounds were identified in the HECS, the main ones being 2-bornanone, menthofuran and camphene. Among the HECS treated animals, all showed significant maximal inhibition (MI) when challenged with pro-inflammatory agents by nebulization. In the group of animals that received CAR, the MI was 72.77 ± 3.88%, compared to the negative control (NC). There was a predominance of lymphocytes (59.18 ± 4.07%). For the HIST group, MI was 64.03 ± 4.33%, in relation to the NC, with predominance in macrophages number (76.53 ± 2.78%). In the CAP group, the MI obtained was 38.90 ± 11.31%, with greater macrophage migration occurring (80.98 ± 3.44%) in the higher dose. For animals subjected to BK inhalation, the MI was 48.63 ± 3.95% with macrophages predominance (64.33 ± 10.04%). In the group that received C48/80 inhalation, the MI was 40.25 ± 5.30% (100 mg kg-1), with higher occurrence of macrophages (72.97 ± 4.23%). CONCLUSION: Our results suggest that HECS had a non-specific inhibitory effect on cellular migration induced by different pro-inflammatory agents, reducing inflammation in airways of mice. These effects coincide and support its ethnopharmacological use as an alternative medicine for respiratory diseases in regions where the plant is prevalent.