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Previous issue date: 2017-06-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Kalanchoe brasiliensis e Kalanchoe pinnata (Crassulaceae), conhecidas como ?sai?o? e ?coirama?, t?m amplo uso popular no tratamento de ?lceras p?pticas e inflama??es cut?neas. Vale destacar que K. pinnata est? presente na Rela??o Nacional de Plantas de Interesse do Sistema ?nico de Sa?de - RENISUS (2009). Dentro deste contexto, o objetivo do presente estudo foi caracterizar os marcadores qu?micos nos sucos das folhas das duas esp?cies e avaliar as atividades gastroprotetora e anti-inflamat?ria t?pica. Foi realizada caracteriza??o fitoqu?mica por Cromatografia em Camada Delgada (CCD) e Cromatografia L?quida de Ultra Efici?ncia Acoplada a Espectr?metro de Massas (CLUE-EM). A atividade gastroprotetora foi avaliada nos modelos de ?lcera ?guda induzida por etanol e indometacina, j? a secre??o g?strica foi avaliada no modelo de ligadura do piloro, em ratos Wistar. Foi realizado o pr?- tratamento com os sucos nas doses de 125, 250 e 500 mg/kg e a ranitidina (50 mg/kg) por via oral. A atividade anti-inflamat?ria t?pica foi avaliada no modelo de edema de pata induzido por carragenina e edema de orelha induzido por ?leo de crot?n em camundongos Swiss, utilizando formula??es na forma de gel contendo os sucos em diferentes concentra??es (1,25%, 2,5% e 5%) e como f?rmaco padr?o a dexametasona (1mg/g), todos administrados por via t?pica imediatamente ap?s a indu??o. A an?lise por CCD revelou a presen?a de manchas caracter?sticas de flavonoides nos sucos das duas esp?cies, ap?s revela??o com o Reagente Natural A, sendo observado que as duas esp?cies t?m perfil flavono?dico diferente. Na an?lise por CLUE-EM K. brasiliensis apresentou flavonoides glicosilados derivados principalmente da patuletina, enquanto que K. pinnata apresentou flavonoides glicosilados derivados principalmente da quercetina. O pr?-tratamento com o suco das folhas de K. brasiliensis nas doses de 125 mg/kg (P<0,01), 250 mg/kg e 500 mg/kg (P<0,001) e K. pinnata nas doses de 125 mg/kg (P<0,01), 250 mg/k e 500 mg/kg (P<0,001) reduziram significativamente as les?es em compara??o ao controle positivo no modelo de indu??o por etanol. No modelo de indu??o por indometacina o suco das folhas de K. brasiliensis apresentou resultado significativo nas doses de 250 (P<0,05) e 500 mg/kg (P<0,01) e K. pinnata nas doses de 250 e 500 mg/kg (P<0,001). A redu??o das les?es foi acompanhada de aumento do conte?do total de glutationa e redu??o dos n?veis de malondialde?do. Al?m disso, houve redu??o dos n?veis de mieloperoxidase, IL-1? e TNF-?. Tamb?m foi observado efeito citoprotetor na avalia??o histol?gica com H&E e manuten??o da produ??o de muco com PAS, al?m da redu??o da express?o de iNOS e NF-?B p65 e aumento da express?o de ZO-1 por imunohistoqu?mica. Os sucos das folhas das duas esp?cies n?o alteraram a acidez, o pH e o volume do suco g?strico. No modelo de edema de orelha, as formula??es contendo as tr?s concentra??es do suco das folhas de K. brasiliensis reduziram significativamente o edema quando comparadas ao grupo placebo (1,25% P<0,05; 2,5% P<0,01 e 5% P<0,01). Entretanto, apenas a formula??o contendo o suco das folhas de K. pinnata na concentra??o de 5% apresentou resultado significativo (P<0,01). No modelo de edema de pata, as formula??es contendo o suco das folhas de K. brasiliensis nas concentra??es de 1,25 e 2,5% reduziram significativamente (P<0,05) o edema no tempo 4 h. A formula??o na concentra??o de 5% reduziu significativamente o edema nos tempos 1 h (P<0,001), 2 h, 3 h e 4 h (P<0,01). Em rela??o as formula??es contendo o suco das folhas de K. pinnata, a concentra??o de 1,25% reduziu significativamente o edema no tempo 1h (P<0,01) e 2h (P<0,05), na concentra??o de 5% reduziu significativamente no tempo 1h (P<0,05). A diminui??o do edema foi acompanhada da redu??o de miloperoxidase. Concluiu-se que os sucos das duas esp?cies apresentaram atividade gastroprotetora e anti-inflamat?ria t?pica em modelos in vivo, resultados que justificam a utiliza??o popular das esp?cies. / Kalanchoe brasiliensis and Kalanchoe pinnata (Crassulaceae), known as "sai?o" and "coirama", have wide popular use in the treatment of peptic ulcers and cutaneous inflammations. It is worth mentioning that K. pinnata is present in the National List of Plants of Interest of the Unified Health System - RENISUS (2009). Within this context, the objective of the present study was to characterize the chemical markers in the leaf juices of both species and to evaluate the gastroprotective and topical anti-inflammatory activities. Phytochemical characterization was performed by Thin Layer Chromatography (TLC) and Ultra High Performance Liquid Chromatography coupled to Mass Spectrometer (UHPLC- MS). Gastroprotective activity was evaluated in ethanol and indomethacin induced acute ulcer models, whereas gastric secretion was evaluated in the pylorus ligature model in Wistar rats. Pre-treatment was performed with the juices at the doses of 125, 250 and 500 mg/kg and ranitidine (50 mg/kg) orally. The topical anti-inflammatory activity was evaluated in the carrageenan induced paw edema model and croton oil-induced ear edema in Swiss mice using gel formulations containing the juices at different concentrations (1,25%, 2,5% and 5%) and as the standard drug dexamethasone (1mg/g), all administered topically immediately after induction. The TLC analysis revealed the presence of flavonoid stains in the juices of both species after revelation with the Natural Reagent A, being observed that the two species have different flavonoid profiles. In the analysis by UHPLC-MS the K. brasiliensis leaf juice showed glycosylated flavonoids derived mainly from patuletin, while that of K. pinnata presented glycosylated flavonoids derived mainly from quercetin. The pre-treatment with the K. brasiliensis leaf juice at doses of 125 mg/kg (P<0,01), 250 mg/kg and 500 mg/kg (P<0,001) and K. pinnata at doses of 125 mg/kg (P<0,01), 250 mg/kg and 500 mg/kg (P<0,001) significantly reduced the lesions compared to the positive control in the ethanol induction model. In the indomethacin induction model, the K. brasiliensis leaf juice showed significant results at doses of 250 mg/kg (P<0,05) and 500 mg/kg (P<0,01) and K. pinnata at doses of 250 and 500 mg/kg (P<0,001). Reduction of lesions was accompanied by an increase in total glutathione content and reduction of malondialdehyde levels. In addition, levels of myeloperoxidase, IL-1? and TNF-? were reduced. Cytoprotective effect was also observed in histological evaluation with H&E and maintenance of mucus production with PAS, as well as reduction of iNOS and NF-?B p65 expression and increased expression of ZO-1 by immunohistochemistry. Leaf juices from both species did not change the acidity, pH and volume of the gastric juice. In the ear edema model, the formulations containing the three concentrations of the K. brasiliensis leaf juice significantly reduced the edema when compared to the placebo group (1,25% P<0,05, 2,5% P<0,01 and 5% P<0,01). However, only the formulation containing the juice of the K. pinnata leaf juice at 5% concentration showed a significant result (P<0,01). In the paw edema model, the formulations containing the K. brasiliensis leaf juice at concentrations of 1,25 and 2,5% significantly reduced (P<0,05) the edema in the time 4 h. The formulation at 5% concentration significantly reduced edema at 1 h (P <0,001), 2 h, 3 h and 4 h (P <0,01). Regarding the formulations containing the K. pinnata leaf juice, the concentration of 1,25% significantly reduced the edema in the time 1 h (P<0,01) and 2 h (P<0,05), in the concentration of 5% significantly reduced in time 1 h (P<0,05). The decrease in edema was followed by reduction of miloperoxidase. It was concluded that the juices of both species presented gastroprotective and topical anti-inflammatory activity in vivo models, results that justify the popular use of the species.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/24572 |
| Date | 30 June 2017 |
| Creators | Ara?jo, Edilane Rodrigues Dantas de |
| Contributors | 81995725072, Ferreira, Leandro de Santis, 21995330850, Batista, Le?nia Maria, 46775943415, Guerra, Gerlane Coelho Bernardo, Langassner, Silvana Maria Zucolotto |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM CI?NCIAS FARMAC?UTICAS, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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