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Hemodynamic Regulation of Endothelial Cell Gene Expression: Effects of p65 Expression Level on Constitutive and TNFα Induced NF-κB SignallingWon, Doyon 28 September 2009 (has links)
Atherosclerosis is a chronic inflammatory disease of arterial blood vessels, characterized by deposition of lipoproteins in the arterial wall. Atherosclerotic plaques form preferentially in distinct regions of the vasculature such as branch points, curvatures and bifurcations, suggesting that local hemodynamic forces may contribute to disease susceptibility. Shear stress imparted on endothelial cells (ECs) by the flowing blood has been shown to modulate gene expression and remodelling of the artery.
In this thesis, an in vitro model was established to recreate the contrasting environments found in atherosclerosis-prone and atherosclerosis-resistant regions of the vasculature to demonstrate a direct causal-relationship between shear stress and expression of endothelial nitric oxide synthase (eNOS) and p65 in ECs. In vitro assessment of cell shape and expression patterns of these anti- and atherogenic genes demonstrated that shear stress can induce cell morphology and gene expression patterns that are similar to ECs in atherosclerosis-prone and atherosclerosis-resistant regions of the mouse vasculature. Regulation of eNOS transcription by shear stress was demonstrated using a transgenic mouse model and in vitro heterogeneous nuclear RNA (hnRNA) quantification.
Similar to ECs in atherosclerosis-prone regions, epithelial cells lining the small intestine lumen express high levels of p65. To investigate the effects of p65 expression levels on constitutive and tumour necrosis factor α (TNFα)-induced nuclear factor-κB (NF-κB) signalling, p65 expression was suppressed in HeLa cells by RNA interference. Lower p65 expression resulted in reduced TNFα-induced expression of NF-κB target genes, including many subunits of inhibitor of nuclear factor κB (IκB), demonstrating modulation of NF-κB priming by p65 expression levels. Suppression of p65 also affected constitutive expression levels of IκB, and resulted in re-setting of the NF-κB/IκB equilibrium. Experiments using inhibitors of canonical NF-κB signalling found that basal expression of NF-κB components is independent of nuclear factor κB kinase β (IKKβ) activity and proteasome-mediated degradation of IκBα. Together, these studies elucidate the mechanism of flow-mediated gene regulation and the effect of resulting changes in p65 expression on NF-κB signalling.
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Effect of protein-energy malnutrition on nuclear factor kappa B activation following global ischemiaJi, Liang 11 December 2006
Our laboratory previously found that protein-energy malnutrition (PEM) existing prior to brain ischemia impaired functional outcome measured in an open field test, and one-third of animals showed a marked increase in reactive gliosis. It was hypothesized that PEM worsened stroke outcome by increasing inflammation via increased activation of the transcription factor, nuclear factor kappa B (NFκB). Mongolian gerbils (11-12 wk old) were randomly assigned to a control diet (12.5% protein) or a protein-deficient diet (2%) for 28 days. The control group on average gained 4.9g and the PEM group lost 7.4g. PEM gerbils had significantly decreased food intake (P<0.001; unpaired t-test). Animals were then subjected to global ischemia or sham surgery, resulting in four experimental groups. Global ischemia was achieved by a 5 min bilateral common carotid artery occlusion with tympanic temperature regulated at 36.5 ± 0.2C. PEM independently increased hippocampal NFκB activation by three times higher than control diet animals at 6hr after surgery (p=0.014; 2-factor ANOVA) detected by electrophoretic mobility shift assay (EMSA). There was no significant effect of ischemia on NFκB activation and there was no interaction of diet and ischemia. Serum glucose and serum cortisol were also measured since both variables can be affected by PEM and can influence stroke outcome, but there was no significant effect of diet or ischemia. Because of the increased NFκB activation observed in PEM-Sham animals, a second experiment investigated if PEM also increased NFκB activation in the absence of surgery. Gerbils of the same age were randomly assigned to either control diet or PEM for 28 days but did not receive any surgery. PEM consistently increased NFκB activation. Since PEM exists in 16% of elderly stroke patients at admission, the data suggest that PEM may worsen stroke outcome through increased activation of NFκB. Because increased NFκB activation was also observed in PEM independent of ischemia, the data also have implications for the inflammatory response of protein-energy malnourished elderly in general.
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Effect of protein-energy malnutrition on nuclear factor kappa B activation following global ischemiaJi, Liang 11 December 2006 (has links)
Our laboratory previously found that protein-energy malnutrition (PEM) existing prior to brain ischemia impaired functional outcome measured in an open field test, and one-third of animals showed a marked increase in reactive gliosis. It was hypothesized that PEM worsened stroke outcome by increasing inflammation via increased activation of the transcription factor, nuclear factor kappa B (NFκB). Mongolian gerbils (11-12 wk old) were randomly assigned to a control diet (12.5% protein) or a protein-deficient diet (2%) for 28 days. The control group on average gained 4.9g and the PEM group lost 7.4g. PEM gerbils had significantly decreased food intake (P<0.001; unpaired t-test). Animals were then subjected to global ischemia or sham surgery, resulting in four experimental groups. Global ischemia was achieved by a 5 min bilateral common carotid artery occlusion with tympanic temperature regulated at 36.5 ± 0.2C. PEM independently increased hippocampal NFκB activation by three times higher than control diet animals at 6hr after surgery (p=0.014; 2-factor ANOVA) detected by electrophoretic mobility shift assay (EMSA). There was no significant effect of ischemia on NFκB activation and there was no interaction of diet and ischemia. Serum glucose and serum cortisol were also measured since both variables can be affected by PEM and can influence stroke outcome, but there was no significant effect of diet or ischemia. Because of the increased NFκB activation observed in PEM-Sham animals, a second experiment investigated if PEM also increased NFκB activation in the absence of surgery. Gerbils of the same age were randomly assigned to either control diet or PEM for 28 days but did not receive any surgery. PEM consistently increased NFκB activation. Since PEM exists in 16% of elderly stroke patients at admission, the data suggest that PEM may worsen stroke outcome through increased activation of NFκB. Because increased NFκB activation was also observed in PEM independent of ischemia, the data also have implications for the inflammatory response of protein-energy malnourished elderly in general.
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Hemodynamic Regulation of Endothelial Cell Gene Expression: Effects of p65 Expression Level on Constitutive and TNFα Induced NF-κB SignallingWon, Doyon 28 September 2009 (has links)
Atherosclerosis is a chronic inflammatory disease of arterial blood vessels, characterized by deposition of lipoproteins in the arterial wall. Atherosclerotic plaques form preferentially in distinct regions of the vasculature such as branch points, curvatures and bifurcations, suggesting that local hemodynamic forces may contribute to disease susceptibility. Shear stress imparted on endothelial cells (ECs) by the flowing blood has been shown to modulate gene expression and remodelling of the artery.
In this thesis, an in vitro model was established to recreate the contrasting environments found in atherosclerosis-prone and atherosclerosis-resistant regions of the vasculature to demonstrate a direct causal-relationship between shear stress and expression of endothelial nitric oxide synthase (eNOS) and p65 in ECs. In vitro assessment of cell shape and expression patterns of these anti- and atherogenic genes demonstrated that shear stress can induce cell morphology and gene expression patterns that are similar to ECs in atherosclerosis-prone and atherosclerosis-resistant regions of the mouse vasculature. Regulation of eNOS transcription by shear stress was demonstrated using a transgenic mouse model and in vitro heterogeneous nuclear RNA (hnRNA) quantification.
Similar to ECs in atherosclerosis-prone regions, epithelial cells lining the small intestine lumen express high levels of p65. To investigate the effects of p65 expression levels on constitutive and tumour necrosis factor α (TNFα)-induced nuclear factor-κB (NF-κB) signalling, p65 expression was suppressed in HeLa cells by RNA interference. Lower p65 expression resulted in reduced TNFα-induced expression of NF-κB target genes, including many subunits of inhibitor of nuclear factor κB (IκB), demonstrating modulation of NF-κB priming by p65 expression levels. Suppression of p65 also affected constitutive expression levels of IκB, and resulted in re-setting of the NF-κB/IκB equilibrium. Experiments using inhibitors of canonical NF-κB signalling found that basal expression of NF-κB components is independent of nuclear factor κB kinase β (IKKβ) activity and proteasome-mediated degradation of IκBα. Together, these studies elucidate the mechanism of flow-mediated gene regulation and the effect of resulting changes in p65 expression on NF-κB signalling.
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Investigation of Bioactive Milk Phospholipid Liposomes and Soy Phospholipid Liposomes on Adipocyte PhysiologyKosmerl, Erica L. January 2019 (has links)
No description available.
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The Effects of Aging on Muscle Loss and Nuclear Factor Kappa-B Levels in Rats Fed a Diet Containing Suboptimal Leucine LevelsKohlen, Corinne Rose 01 January 2009 (has links)
Loss of muscle due to aging is often associated with significant detrimental effects. Therefore, it is crucial to understand signaling molecules that may trigger the muscle loss or prevent the process. The transcription factor, Nuclear Factor Kappa-B (NF-κB), is associated with both catabolic and anabolic pathways of muscle metabolism and may be involved in age-related muscle loss. Leucine is an essential amino acid that is required for both protein synthesis and intracellular signaling pathways that regulate protein synthesis and degradation. The current study examined muscle NF-kB levels in male Sprague-Dawley rats, aged 6 (adult) and 21 months (old) fed a diet containing suboptimal leucine levels for 10-17 days. We found that old rats consumed less grams of food per body weight (BW) each day than adult rats (1.45% g diet/g BW vs. 2.4% g diet/g BW). Weight loss during the study was not significantly different between age groups. However the average mass of gastrocnemius and soleus muscles (g muscle/g BW) was significantly lower in old rats. Reduction in gastrocnemius (g muscle/g BW*10²) was associated with 1.8 fold higher muscle cell NF-κB in old vs. adult rats (p = 0.0443). There was also a higher level of ubiquitinated proteins in old gastrocnemius muscle cells relative to the adult gastrocnemius, however differences did not reach statistical significance. For tibialis anterior muscle, the average mass (g muscle/g BW*10²), NF-κB levels and ubiquitinated proteins were not significantly different between adult and old rats. Our findings suggest that aging affects muscle loss and NF-kB in a tissue-specific manner in rats fed a diet with suboptimal leucine levels.
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Efeitos da atorvastatina sobre a ossificação endocondral de fêmures, remodelação óssea e movimentação dentária induzida, estudo em ratosDolci, Gabriel Schmidt January 2016 (has links)
As estatinas são medicamentos comumente prescritos para a prevenção da hiper-lipidemia. Além da redução do colesterol, tais medicamentos parecem estimular a osteogênese e suprimir a reabsorção óssea, o que poderia afetar a movimentação dentária induzida (MDI) e a recidiva ortodôntica. Assim, o objetivo deste estudo foi determinar se a atorvastatina (ATV) pode afetar a MDI, a recidiva e a osteoclastogênese, por meio da modulação da expressão das moléculas: - ligante do receptor ativador de NFκB (RANKL) e osteoprotegerina (OPG). Ainda foram analisados os potenciais efeitos adversos da ATV sobre a ossificação endocondral e sobre o turnover de ossos longos. No primeiro experimento, 36 ratos foram sujeitos a MDI durante 21 dias, quando o aparelho foi removido. Aos animais, foram administrados, diariamente, ATV ou solução salina (SAL), via gavagem. Após 7, 14 e 21 dias de administração de ATV / SAL, a recidiva dentária foi mensurada, e foram obtidos os cortes histológicos da maxila e fêmur, os quais foram submetidos às seguintes colorações: - H&E – para análise histomorfométrica; - fosfatase acida tartrato resistente (TRAP) – para contagem de osteoclastos e; - imunohistoquímica para RANKL e OPG. A atorvastatina resultou numa inibição da recidiva ortodôntica (p < 0,05), e numa transiente redução do número de osteoclastos (p < 0,05); havendo uma correlação positiva e significativa (p < 0,01) entre o estes dois fatores (número de osteoclastos e a taxa de recidiva). A administração de estatinas também aumentou significativamente a expressão de OPG (p < 0,01), mas não a de RANKL. Além disso, após 21 dias de administração de ATV, a espessura da cartilagem da placa de crescimento e da zona hipertrófica condrocítica foi significativamente aumentada. Já no segundo experimento, 24 ratos começaram a receber diariamente ATV ou solução salina (SAL), via gavagem. Duas semanas mais tarde, a MDI foi iniciada. O deslocamento do dente foi medido após 7, 14 e 21 dias, enquanto que os cortes histológicos da maxila e do fêmur foram obtidos após 14 e 21 dias de MDI; sendo então submetidos às colorações de H&E e TRAP, para avaliação histomorfométrica e contagem de osteoclastos. A administração de atorvastatina gerou um menor movimento dentário (p <0,05) e uma redução transitória do número de osteoclastos (p <0,05). No grupo SAL, após 14 dias de MDI, ocorreu um aumento no número de osteoclastos, assim reduzindo a taxa de volume ósseo, quando comparado com as maxilas controle (sem movimento dentário), deste mesmo grupo. Contudo, tal comportamento não foi observado no grupo ATV. Interessantemente, depois de 35 dias, a atorvastatina não afetou a remodelação óssea nas maxilas controle, nem a ossificação endocondral em fêmures. Logo, guardando as devidas limitações deste estudo pré-clínico, nossos resultados sugerem que a administração sistêmica de atorvastatina é capaz de minimizar a MDI e recidiva ortodôntica. No entanto, os seus efeitos celulares sobre a ossificação endocondral e remodelação óssea durante a MDI e recidiva, parecem ser limitados a um curto período de tempo, o que aparentemente necessita de investigações futuras. Finalmente, nossos resultados lançam luz sobre a superexpressão OPG induzida por estatinas, o que representa um alvo molecular para modular o metabolismo do ósseo e, assim minimizar a recidiva ortodôntica. / Statins are drugs commonly prescribed for prevention of hyper-lipidemia. In addition to the cholesterol-lowering, these medicines seem to enhance osteogenesis and suppress bone resorption, which could affect orthodontic tooth movement (OTM) and relapse. Therefore, the aim of this study was to determine whether atorvastatin (ATV) might affect the orthodontic relapse or tooth movement and osteoclastogenesis, through the modulation of the following molecules: receptor activator of nuclear κ B ligand (RANKL) and; - osteoprotegerin (OPG). Furthermore, we analyzed potential adverse effects of ATV on long bone turnover and endochondral ossification. In the first experiment, 36 rats were subjected to OTM for 21 days, when the appliance was removed. After, the animals were administered daily with ATV (15mg/Kg) or saline (SAL), via gavage (n=18, per group). Up to 7, 14 and 21 days of ATV/SAL administration the tooth relapse was measured while maxillary and femur histologic sections were obtained and prepared to: - H&E staining – used in histomorphometric analysis, tartrate resistant acid phosphatase histochemical staining (TRAP) – used to osteoclasts counting and, immunohistochemistry to RANKL and OPG. Atorvastatin resulted in a decreased tooth relapse (p<0.05), and a transient reduction of osteoclasts number (p<0.05). There was a positive and significant correlation (p<0.01) between these two parameters (osteoclasts number and relapse rate). The statin administration increased significantly the OPG (p<0.01), but not the RANKL expression. Furthermore, after 21 days of ATV administration, the thickness of growth plate cartilage and chondrocytic hypertrophic zone was enhanced. In the second experiment, 24 rats started to be administered daily with ATV or SAL, via gavage. Two weeks later, the OTM started. The tooth displacement was measured after 7, 14, and 21 days, while the maxillary and femur histologic sections were obtained only after 14 and 21 days of OTM. At these times, the sections were prepared to H&E and TRAP, intending to perform the histomorphometric analysis and osteoclasts count. Atorvastatin administration promoted a decreased tooth movement (p<0.05), and a transient reduction of osteoclasts number (p<0.05). In the SAL group, after 14 days of OTM, the increased number of osteoclasts was associated to a reduced bone volume rate, when compared to its control maxillae. However, this trend was not obvious in ATV group. Interestingly, after 35 days, statins did not affect the bone turnover and endochondral ossification. The big picture of our study suggests that systemic administration of statins is able to minimize OTM and relapse. However, its cellular effects on endochondral ossification and bone turnover during OTM or relapse seem to be limited to a short period, apparently requiring further investigations. Finally, our results shed light on OPG overexpression induced by statins, which represents a molecular target modulating maxillary bone metabolism thus inhibiting orthodontic relapse.
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FUNCTION OF MYELOID DENDRITIC CELLSZhao,Li Unknown Date
No description available.
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Efeitos da atorvastatina sobre a ossificação endocondral de fêmures, remodelação óssea e movimentação dentária induzida, estudo em ratosDolci, Gabriel Schmidt January 2016 (has links)
As estatinas são medicamentos comumente prescritos para a prevenção da hiper-lipidemia. Além da redução do colesterol, tais medicamentos parecem estimular a osteogênese e suprimir a reabsorção óssea, o que poderia afetar a movimentação dentária induzida (MDI) e a recidiva ortodôntica. Assim, o objetivo deste estudo foi determinar se a atorvastatina (ATV) pode afetar a MDI, a recidiva e a osteoclastogênese, por meio da modulação da expressão das moléculas: - ligante do receptor ativador de NFκB (RANKL) e osteoprotegerina (OPG). Ainda foram analisados os potenciais efeitos adversos da ATV sobre a ossificação endocondral e sobre o turnover de ossos longos. No primeiro experimento, 36 ratos foram sujeitos a MDI durante 21 dias, quando o aparelho foi removido. Aos animais, foram administrados, diariamente, ATV ou solução salina (SAL), via gavagem. Após 7, 14 e 21 dias de administração de ATV / SAL, a recidiva dentária foi mensurada, e foram obtidos os cortes histológicos da maxila e fêmur, os quais foram submetidos às seguintes colorações: - H&E – para análise histomorfométrica; - fosfatase acida tartrato resistente (TRAP) – para contagem de osteoclastos e; - imunohistoquímica para RANKL e OPG. A atorvastatina resultou numa inibição da recidiva ortodôntica (p < 0,05), e numa transiente redução do número de osteoclastos (p < 0,05); havendo uma correlação positiva e significativa (p < 0,01) entre o estes dois fatores (número de osteoclastos e a taxa de recidiva). A administração de estatinas também aumentou significativamente a expressão de OPG (p < 0,01), mas não a de RANKL. Além disso, após 21 dias de administração de ATV, a espessura da cartilagem da placa de crescimento e da zona hipertrófica condrocítica foi significativamente aumentada. Já no segundo experimento, 24 ratos começaram a receber diariamente ATV ou solução salina (SAL), via gavagem. Duas semanas mais tarde, a MDI foi iniciada. O deslocamento do dente foi medido após 7, 14 e 21 dias, enquanto que os cortes histológicos da maxila e do fêmur foram obtidos após 14 e 21 dias de MDI; sendo então submetidos às colorações de H&E e TRAP, para avaliação histomorfométrica e contagem de osteoclastos. A administração de atorvastatina gerou um menor movimento dentário (p <0,05) e uma redução transitória do número de osteoclastos (p <0,05). No grupo SAL, após 14 dias de MDI, ocorreu um aumento no número de osteoclastos, assim reduzindo a taxa de volume ósseo, quando comparado com as maxilas controle (sem movimento dentário), deste mesmo grupo. Contudo, tal comportamento não foi observado no grupo ATV. Interessantemente, depois de 35 dias, a atorvastatina não afetou a remodelação óssea nas maxilas controle, nem a ossificação endocondral em fêmures. Logo, guardando as devidas limitações deste estudo pré-clínico, nossos resultados sugerem que a administração sistêmica de atorvastatina é capaz de minimizar a MDI e recidiva ortodôntica. No entanto, os seus efeitos celulares sobre a ossificação endocondral e remodelação óssea durante a MDI e recidiva, parecem ser limitados a um curto período de tempo, o que aparentemente necessita de investigações futuras. Finalmente, nossos resultados lançam luz sobre a superexpressão OPG induzida por estatinas, o que representa um alvo molecular para modular o metabolismo do ósseo e, assim minimizar a recidiva ortodôntica. / Statins are drugs commonly prescribed for prevention of hyper-lipidemia. In addition to the cholesterol-lowering, these medicines seem to enhance osteogenesis and suppress bone resorption, which could affect orthodontic tooth movement (OTM) and relapse. Therefore, the aim of this study was to determine whether atorvastatin (ATV) might affect the orthodontic relapse or tooth movement and osteoclastogenesis, through the modulation of the following molecules: receptor activator of nuclear κ B ligand (RANKL) and; - osteoprotegerin (OPG). Furthermore, we analyzed potential adverse effects of ATV on long bone turnover and endochondral ossification. In the first experiment, 36 rats were subjected to OTM for 21 days, when the appliance was removed. After, the animals were administered daily with ATV (15mg/Kg) or saline (SAL), via gavage (n=18, per group). Up to 7, 14 and 21 days of ATV/SAL administration the tooth relapse was measured while maxillary and femur histologic sections were obtained and prepared to: - H&E staining – used in histomorphometric analysis, tartrate resistant acid phosphatase histochemical staining (TRAP) – used to osteoclasts counting and, immunohistochemistry to RANKL and OPG. Atorvastatin resulted in a decreased tooth relapse (p<0.05), and a transient reduction of osteoclasts number (p<0.05). There was a positive and significant correlation (p<0.01) between these two parameters (osteoclasts number and relapse rate). The statin administration increased significantly the OPG (p<0.01), but not the RANKL expression. Furthermore, after 21 days of ATV administration, the thickness of growth plate cartilage and chondrocytic hypertrophic zone was enhanced. In the second experiment, 24 rats started to be administered daily with ATV or SAL, via gavage. Two weeks later, the OTM started. The tooth displacement was measured after 7, 14, and 21 days, while the maxillary and femur histologic sections were obtained only after 14 and 21 days of OTM. At these times, the sections were prepared to H&E and TRAP, intending to perform the histomorphometric analysis and osteoclasts count. Atorvastatin administration promoted a decreased tooth movement (p<0.05), and a transient reduction of osteoclasts number (p<0.05). In the SAL group, after 14 days of OTM, the increased number of osteoclasts was associated to a reduced bone volume rate, when compared to its control maxillae. However, this trend was not obvious in ATV group. Interestingly, after 35 days, statins did not affect the bone turnover and endochondral ossification. The big picture of our study suggests that systemic administration of statins is able to minimize OTM and relapse. However, its cellular effects on endochondral ossification and bone turnover during OTM or relapse seem to be limited to a short period, apparently requiring further investigations. Finally, our results shed light on OPG overexpression induced by statins, which represents a molecular target modulating maxillary bone metabolism thus inhibiting orthodontic relapse.
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10 |
Efeitos da atorvastatina sobre a ossificação endocondral de fêmures, remodelação óssea e movimentação dentária induzida, estudo em ratosDolci, Gabriel Schmidt January 2016 (has links)
As estatinas são medicamentos comumente prescritos para a prevenção da hiper-lipidemia. Além da redução do colesterol, tais medicamentos parecem estimular a osteogênese e suprimir a reabsorção óssea, o que poderia afetar a movimentação dentária induzida (MDI) e a recidiva ortodôntica. Assim, o objetivo deste estudo foi determinar se a atorvastatina (ATV) pode afetar a MDI, a recidiva e a osteoclastogênese, por meio da modulação da expressão das moléculas: - ligante do receptor ativador de NFκB (RANKL) e osteoprotegerina (OPG). Ainda foram analisados os potenciais efeitos adversos da ATV sobre a ossificação endocondral e sobre o turnover de ossos longos. No primeiro experimento, 36 ratos foram sujeitos a MDI durante 21 dias, quando o aparelho foi removido. Aos animais, foram administrados, diariamente, ATV ou solução salina (SAL), via gavagem. Após 7, 14 e 21 dias de administração de ATV / SAL, a recidiva dentária foi mensurada, e foram obtidos os cortes histológicos da maxila e fêmur, os quais foram submetidos às seguintes colorações: - H&E – para análise histomorfométrica; - fosfatase acida tartrato resistente (TRAP) – para contagem de osteoclastos e; - imunohistoquímica para RANKL e OPG. A atorvastatina resultou numa inibição da recidiva ortodôntica (p < 0,05), e numa transiente redução do número de osteoclastos (p < 0,05); havendo uma correlação positiva e significativa (p < 0,01) entre o estes dois fatores (número de osteoclastos e a taxa de recidiva). A administração de estatinas também aumentou significativamente a expressão de OPG (p < 0,01), mas não a de RANKL. Além disso, após 21 dias de administração de ATV, a espessura da cartilagem da placa de crescimento e da zona hipertrófica condrocítica foi significativamente aumentada. Já no segundo experimento, 24 ratos começaram a receber diariamente ATV ou solução salina (SAL), via gavagem. Duas semanas mais tarde, a MDI foi iniciada. O deslocamento do dente foi medido após 7, 14 e 21 dias, enquanto que os cortes histológicos da maxila e do fêmur foram obtidos após 14 e 21 dias de MDI; sendo então submetidos às colorações de H&E e TRAP, para avaliação histomorfométrica e contagem de osteoclastos. A administração de atorvastatina gerou um menor movimento dentário (p <0,05) e uma redução transitória do número de osteoclastos (p <0,05). No grupo SAL, após 14 dias de MDI, ocorreu um aumento no número de osteoclastos, assim reduzindo a taxa de volume ósseo, quando comparado com as maxilas controle (sem movimento dentário), deste mesmo grupo. Contudo, tal comportamento não foi observado no grupo ATV. Interessantemente, depois de 35 dias, a atorvastatina não afetou a remodelação óssea nas maxilas controle, nem a ossificação endocondral em fêmures. Logo, guardando as devidas limitações deste estudo pré-clínico, nossos resultados sugerem que a administração sistêmica de atorvastatina é capaz de minimizar a MDI e recidiva ortodôntica. No entanto, os seus efeitos celulares sobre a ossificação endocondral e remodelação óssea durante a MDI e recidiva, parecem ser limitados a um curto período de tempo, o que aparentemente necessita de investigações futuras. Finalmente, nossos resultados lançam luz sobre a superexpressão OPG induzida por estatinas, o que representa um alvo molecular para modular o metabolismo do ósseo e, assim minimizar a recidiva ortodôntica. / Statins are drugs commonly prescribed for prevention of hyper-lipidemia. In addition to the cholesterol-lowering, these medicines seem to enhance osteogenesis and suppress bone resorption, which could affect orthodontic tooth movement (OTM) and relapse. Therefore, the aim of this study was to determine whether atorvastatin (ATV) might affect the orthodontic relapse or tooth movement and osteoclastogenesis, through the modulation of the following molecules: receptor activator of nuclear κ B ligand (RANKL) and; - osteoprotegerin (OPG). Furthermore, we analyzed potential adverse effects of ATV on long bone turnover and endochondral ossification. In the first experiment, 36 rats were subjected to OTM for 21 days, when the appliance was removed. After, the animals were administered daily with ATV (15mg/Kg) or saline (SAL), via gavage (n=18, per group). Up to 7, 14 and 21 days of ATV/SAL administration the tooth relapse was measured while maxillary and femur histologic sections were obtained and prepared to: - H&E staining – used in histomorphometric analysis, tartrate resistant acid phosphatase histochemical staining (TRAP) – used to osteoclasts counting and, immunohistochemistry to RANKL and OPG. Atorvastatin resulted in a decreased tooth relapse (p<0.05), and a transient reduction of osteoclasts number (p<0.05). There was a positive and significant correlation (p<0.01) between these two parameters (osteoclasts number and relapse rate). The statin administration increased significantly the OPG (p<0.01), but not the RANKL expression. Furthermore, after 21 days of ATV administration, the thickness of growth plate cartilage and chondrocytic hypertrophic zone was enhanced. In the second experiment, 24 rats started to be administered daily with ATV or SAL, via gavage. Two weeks later, the OTM started. The tooth displacement was measured after 7, 14, and 21 days, while the maxillary and femur histologic sections were obtained only after 14 and 21 days of OTM. At these times, the sections were prepared to H&E and TRAP, intending to perform the histomorphometric analysis and osteoclasts count. Atorvastatin administration promoted a decreased tooth movement (p<0.05), and a transient reduction of osteoclasts number (p<0.05). In the SAL group, after 14 days of OTM, the increased number of osteoclasts was associated to a reduced bone volume rate, when compared to its control maxillae. However, this trend was not obvious in ATV group. Interestingly, after 35 days, statins did not affect the bone turnover and endochondral ossification. The big picture of our study suggests that systemic administration of statins is able to minimize OTM and relapse. However, its cellular effects on endochondral ossification and bone turnover during OTM or relapse seem to be limited to a short period, apparently requiring further investigations. Finally, our results shed light on OPG overexpression induced by statins, which represents a molecular target modulating maxillary bone metabolism thus inhibiting orthodontic relapse.
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