Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-11-06T21:13:15Z No. of bitstreams: 1 MardemPortelaEVasconcelosBarreto_DISSERT.pdf: 1565077 bytes, checksum: 5553fd2358ea31595fb86bc2f4686161 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-11-20T23:45:30Z (GMT) No. of bitstreams: 1 MardemPortelaEVasconcelosBarreto_DISSERT.pdf: 1565077 bytes, checksum: 5553fd2358ea31595fb86bc2f4686161 (MD5) / Made available in DSpace on 2017-11-20T23:45:30Z (GMT). No. of bitstreams: 1 MardemPortelaEVasconcelosBarreto_DISSERT.pdf: 1565077 bytes, checksum: 5553fd2358ea31595fb86bc2f4686161 (MD5) Previous issue date: 2017-07-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O diabetes mellitus causa uma s?rie de complica??es sist?micas, incluindo o retardo no processo de cicatriza??o. Diversas alternativas terap?uticas t?m sido testadas em estudos in vitro e in vivo para promover melhora no processo de reparo de feridas em animais e indiv?duos diab?ticos. O objetivo do presente estudo foi avaliar o efeito da associa??o da laserterapia de baixa intensidade com a administra??o t?pica de pr?polis verde em feridas cut?neas em ratos com diabetes induzida por estreptozotocina. Foram utilizados 90 animais, alocados nos seguintes grupos: (N) normoglic?micos (sem indu??o), sem terapia; (C) controle diab?tico, sem terapia; (L) submetidos ? laserterapia de baixa intensidade (660 nm, 30 mW, 4 J/cm2); (P) submetidos ? administra??o t?pica de pr?polis verde (extrato alco?lico a 30%); e (LP) submetidos ? combina??o de laserterapia com administra??o t?pica de pr?polis verde. Os procedimentos terap?uticos foram realizados a cada 24 horas, por 6 dias. As ?reas cir?rgicas foram fotografadas em dias intercalados, para avalia??o da ?rea de fechamento da ferida. Nos intervalos de 7, 14 e 21 dias parte dos animais foi submetida ? eutan?sia e posterior remo??o da ?rea da ferida. Os esp?cimes foram fixados, processados rotineiramente e inclu?dos em parafina e as l?minas obtidas foram coradas por H/E e Picrosirius red, para avalia??o da reepiteliza??o, intensidade do infiltrado inflamat?rio e forma??o e organiza??o de col?geno, al?m da imunomarca??o para FGF-2 e VEGF. Os dados quantitativos foram submetidos a testes estat?sticos n?o param?tricos, com intervalo de confian?a de 95%. A avalia??o macrosc?pica da ?rea da ferida mostrou que os tr?s grupos tratados (L, P e LP) exibiram uma acelera??o da retra??o da ferida em rela??o ao grupo C (p<0,001) a partir do 3? at? o 14? dia, com resultado semelhante ao grupo N. O grupo LP apresentou um melhor resultado em rela??o aos demais (p<0,05) a partir do 5? dia. A an?lise histol?gica mostrou que os grupos tratados exibiram maiores ?ndices de reepiteliza??o, especialmente nos grupos L e LP, e tamb?m menores ?ndices de inflama??o, com destaque para os grupos LP e P. Com rela??o ? propor??o col?geno I/III observou-se no 7? dia valores maiores no grupo LP em rela??o ao grupo C (p<0.05), assemelhando-se ao grupo N. No 14? dia o grupo L apresentou a maior propor??o dos grupos tratados, aproximando-se dos resultados do grupo N e com diferen?a estat?stica para os demais grupos experimentais (p<0,01). N?o houve diferen?a na propor??o col?geno I/III entre os grupos no intervalo de 21 dias. Os tr?s grupos tratados exibiram menor express?o de FGF-2 e VEGF em compara??o com os grupos N e C, especialmente o grupo LP (p<0,05). Em conjunto, os resultados do presente trabalho permitem concluir que a associa??o da laserterapia com a aplica??o t?pica de pr?polis acelera o processo e a qualidade do reparo no modelo animal de diabetes estudado. / Diabetes mellitus causes a number of systemic complications, including delay in the healing process. Several therapeutic alternatives have been tested in in vitro and in vivo studies in order to promote improvement in the process of wound repair in diabetic animals and individuals. The aim of the present study was to evaluate the effect of the association of low intensity laser therapy and topical administration of green propolis in cutaneous wounds model in rats with streptozotocin-induced diabetes. Ninety animals were allocated in the following groups: (N) normoglycemic (not induced), without therapy; (C) diabetic control, without therapy; (L) undergoing low intensity laser therapy (660 nm, 30 mW, 4 J/cm2); (P) submitted to topical administration of green propolis (30% alcoholic extract); and (LP) submitted to the combination of laser therapy and topical administration of green propolis. Therapeutic procedures were performed every 24 hours for 6 days. The surgical areas were photographed to evaluate the wound closure area. At intervals of 7, 14 and 21 days, the animals were submitted to euthanasia and subsequent removal of the wound area. The specimens were fixed, routinely processed, and embedded in paraffin and the slides obtained were stained by H/E and Picrosirius red for evaluation of re-epithelization, intensity of inflammatory infiltrate, and formation and organization of collagen, and also submitted to immunostaining for FGF-2 and VEGF. The quantitative data were submitted to non-parametric statistical tests, with a 95% confidence interval. The macroscopic evaluation of the wound area showed that the three groups submitted to treatment (L, P, and LP) showed an acceleration of wound retraction in relation to group C (p <0.001) from the 3rd to the 14th day, with results similar to group N. The LP group presented a better result in relation to the others (p <0.05) from the 5th day onwards. Histological analysis showed that the treated groups exhibited higher rates of re-epithelialization, especially in the L and LP groups, as well as lower inflammation rates, especially in the LP and P groups. Regarding the collagen I/III ratio, it was observed on the 7th day higher values in the LP group compared to the C group (p <0.05), similar to the N group. On the 14th day, the L group presented the highest proportion of the treated groups, resembling to the results of group N and with statistical difference for the other experimental groups (p <0.01). There was no difference in the collagen I/III ratio among the groups at the 21-day interval. The three treated groups (L, P, and LP) exhibited lower expression of FGF-2 and VEGF compared to N and C groups, especially the LP group (p <0.05). Taken together, the results of the present study allow us to conclude that the association of laser therapy with the topical application of propolis improves the cutaneous wound repair in the animal model of diabetes studied.

Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Propolis is a resinous material collected by bees from different parts of plants. Both its composition as its biological properties are dependent on factors such as climate, soil, vegetation and species of collecting bees. Propolis has great potential for therapeutic use due to its chemical composition and pharmacological properties. Herein it was determined the contents of phenolic compounds, antioxidant capacity and chemical composition (assessed by HPLC-PAD) of twelve green propolis samples from beekeepers and sixteen commercial propolis extracs from different regions of Southeast Brazil. The antioxidant abilities of the extracts were qualitatively determined through the total phenolic contents using the Folin?Ciocalteau method. Total flavonoids were assessed by the method of complexation with aluminium chloride. The quantitative antioxidant activiti of propolis extracts were determined both by trapping the organic radicals DPPH and ABTS.+ as well by the iron reduction method (FRAP). High pressure liquid chromatography allowed the identification and quantification of chlorogenic acid, caffeic acid, ferulic acid, para-coumaric acid, rosmarinic acid, vanillin, hesperidin, naringenin, pinobanksin, kaempferol, 4-hydroxy-3,5-diprenyl cinnamic acid (Artepillin-C), kampheride and pinostrobin. Artepillin C, a well known chemical marker of propolis, was present in all propolis extracts analyzed. We highlight that the presence of rosmarinic acid in propolis samples from Rio de Janeiro was reported for the first time in this work. The discrimination between green propolis in natura and commercial green propolis extracts was performed using PCA (Principal Component Analysis) statistical method. The antioxidant capacity in vivo of propolis extracts were evaluated using Saccharomyces cerevisiae as biological system model, assessing important parameters as stress tolerance and lipid peroxidation rate. The antifungal activity of ethanol extracts of propolis in natura were tested against the phytopathogenic fungus Fusarium oxysporium. / A pr?polis ? um material resinoso coletado pelas abelhas de diferentes partes das plantas, sua composi??o e as suas propriedades biol?gicas dependem do clima, solo, vegeta??o e da esp?cie da abelha. A pr?polis tem grande potencial de aplica??o terap?utica, devido ? sua composi??o e propriedades farmacol?gicas. Nesse trabalho foi determinado o teor de subst?ncias fen?licas, a capacidade antioxidante e composi??o qu?mica por CLAE-DAD de doze amostras de pr?polis verde, adquiridas de apicultores, e dezesseis extratos de pr?polis comerciais, provenientes de diferentes regi?es do Sudeste do Brasil. A capacidade antioxidante dos extratos foi avaliada qualitativamente atrav?s do teor de fen?licos totais, pelo m?todo de Folin?Ciocalteau, e flavonoides pelo m?todo de complexa??o com cloreto de alum?nio. A quantifica??o do potencial antioxidante foi realizada pela captura dos radicais org?nicos DPPH e ABTS.+, al?m do m?todo de redu??o do ?on f?rrico (FRAP). An?lises por cromatografia l?quida de alta efici?ncia permitiram a identifica??o e quantifica??o de ?cido clorog?nico, ?cido cafeico, ?cido fer?lico, ?cido para-cum?rico, ?cido rosmar?nico, ?cido 3,5-diprenil-4-hidroxicin?mico (Artepillin C), vanilina, hesperidina, naringenina, pinobanksina, canferol, canferide e pinostrobina. Em todos os extratos de pr?polis analisados foi identificado o Artepillin C, marcador qu?mico para pr?polis verde. Destacamos que neste trabalho foi reportada pela primeira vez a presen?a de ?cido rosmar?nico em pr?polis do Rio de Janeiro. M?todos quimiom?tricos de an?lise explorat?ria, utilizando ?nalise por Componentes Principais foram utilizados para discriminar extratos de pr?polis verde in natura daqueles extratos comerciais. O potencial antioxidante in vivo dos extratos etan?licos de pr?polis foram avaliados utilizando cepas controles de Saccharomyces cerevisiae como modelo de sistema biol?gico e foram avaliados toler?ncia ao estresse e proxida??o lip?dica. A atividade antif?ngica dos extratos etan?licos de pr?polis in natura foram avaliados, e inibiram o crescimento in vitro do fungo fitopat?geno Fusarium oxysporum.