EFEITO DE Morinda citrifolia NA MATURAÇÃO IN VITRO DE OÓCITOS BOVINOS

The in vitro production (IVP) of embryos is a biotechnology that helps the production of genetically superior animals. However, the excessive production of oxygen-reactive species by this technique results in cellular damage to oocytes and embryos. The use of antioxidants during IVP can reduce oxidative stress. The fruit of Morinda citrifolia (noni) may cause positive impacts on the maturation rate of the IVP technique due to its antioxidant properties. Thus, this study aims to evaluate the effects of noni on cellular and molecular aspects in the stage of in vitro maturation (IVM) of bovine oocytes. A total of 817 viable immature oocytes were obtained, which were distributed according to the following treatments with noni: control, 1mg/mL, mg/mL, and 10mg/mL. The rate of matured oocytes was determined by the Hoescht 33342 technique. Some oocytes were used to measure the expression of genes related to apoptosis, antioxidant action, and oxidative stress through the qRT-PCR technique. Few oocytes in germinal vesicle and germinal vesicle rupture stage were obtained; however, a large number of oocytes without identifiable nuclear stage and degenerated were obtained. There was no difference for in nuclear findings between the groups treated with noni. The molecular analysis revealed differences between the experimental groups for the Bcl-2 and BAX genes, indicating that the treatment with noni played an important role in anti-apoptotic protection during IVM, mainly evidenced by the expression of mRNA from the 1 mg/mL noni group. Therefore, research data indicate that Morinda citrifolia had an anti-apoptotic effect during in vitro maturation.A produção in vitro (PIV) de embriões é uma biotecnologia que auxilia a produção de animais geneticamente superiores. Contudo, a produção excessiva das espécies reativas de oxigênio por esta técnica resulta em danos celulares aos oócitos e embriões. O uso de antioxidantes durante a PIV pode reduzir o estresse oxidativo. O fruto da Morinda citrifolia (noni) pode causar impactos positivos nas taxas de maturação da técnica da PIV devido suas propriedades antioxidantes. Desta forma, este estudo visa avaliar os efeitos do noni sobre os aspectos celulares e moleculares da etapa de maturação in vitro (MIV) de oócitos bovinos. Obteve-se um total de 817 complexos cumulus oophorus (CCOs) viáveis para a maturação, os quais foram distribuídos de acordo com os seguintes tratamentos com noni: controle, 1mg/mL, 5mg/mL and 10mg/mL. Determinou-se a taxa de oócitos maturados pela técnica Hoescht 33342. Alguns oócitos foram utilizados para mensurar a expressão de genes relacionados a apoptose, ação antioxidante e estresse oxidativo através de qRT-PCR. Foram obtidos poucos oócitos em estágio de vesícula germinativa e rompimento de vesícula germinativa, contudo, obteve-se um grande número de oócitos sem estágio nuclear identificável e degenerados. Não houve diferença para os achados nucleares entre os grupos tratados com noni. A análise molecular revelou diferença entre os grupos experimentais para os genes Bcl-2 e BAX, indicando que o tratamento com noni desempenhou papel importante de proteção antiapoptótica durante a MIV, evidenciado principalmente pela expressão de mRNA do grupo noni 1 mg/mL. Desta forma, os dados da pesquisa indicam que a M. citrifolia apresentou efeito antiapoptótico durante maturação in vitro

Sulforaphane Modulates Joint Inflammation in a Murine Model of Complete Freund’s Adjuvant-Induced Mono-Arthritis

Rheumatoid arthritis (RA) is characterized by inflammation of one or more joints, and affects ~1% of the adult population worldwide. Sulforaphane (SFN) is a natural compound that has been suggested as an antioxidant. Here, SFN’s effects were evaluated in a murine mono-arthritis model. Mono-arthritis was induced in mice by a single intra-articular injection of Complete Freund’s Adjuvant (CFA-10 µg/joint, in 10 µL) into the ipsilateral joint. The contralateral joint received an equal volume of PBS. On the 4th day post-joint inflammation induction, animals received either SFN (10 mg/kg) or vehicle (3% DMSO in saline), intraperitoneally (i.p.), twice a day for 3 days. Joint swelling and secondary mechanical allodynia and hyperalgesia were evaluated over 7 days post-CFA. After this period, animals were culled and their blood and synovial fluid samples were collected for analysis of cell populations, cytokine release and thioredoxin reductase (TrxR) activity. Knee joint samples were also collected for histology. SFN reduced joint swelling and damage whilst increasing the recruitment of Ly6C+ and Ly6G+ cells to CFA-injected joints. SFN-treated animals presented down-regulation of CD11b and CD62L on synovial fluid Ly6G+ cells. Synovial fluid samples obtained from CFA-injected joints and plasma samples of SFN-treated mice presented higher levels of IL-6 and increased activity of TrxR, in comparison with controls. These results indicate that SFN reduces knee joint damage by modulating cell activation/migration to the joints, cytokine production and increasing the activity of TrxR, and therefore, may represent an alternative treatment to joint inflammation