Efeitos de limosilactobacillus reuteri associado à vitamina K2 em camundongos ovariectomizados
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Data
2023-05-09
Autores
Santos, Thaís Aguiar
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Universidade Estadual Paulista (Unesp)
Resumo
A osteoporose é uma doença osteometabólica, caracterizada pelo desequilíbrio entre deposição e reabsorção óssea. Probióticos e vitamina K2 (MK-7) têm recebido atenção como terapia para a osteoporose, isoladamente. A associação de probióticos com a vitamina K é interessante, visto que suas vias benéficas possivelmente estão interligadas, podendo apresentar efeitos sinérgicos. Neste estudo, foram avaliados os efeitos de Limosilactobacillus reuteri associado à vitamina K2 (MK-7) na perda óssea induzida por ovariectomia em camundongos e no crescimento bacteriano, in vitro. No estudo in vivo, os animais foram distribuídos em cinco grupos: falsa OVX (controle); ovariectomia (OVX); OVX + L. reuteri (LR); OVX + vitamina K2 (K); OVX + LR + K. Após 4 semanas de tratamento, avaliaram-se as propriedades biomecânicas de fêmur, microarquitetura vertebral (L5), níveis de osteocalcina carboxilada e não carboxilada, expressão gênica de Ocludina, Jam3, e altura de vilos e criptas intestinais. O tratamento com L. reuteri, MK-7, assim como a sua associação, melhoraram significativamente as propriedades biomecânicas do fêmur, deterioradas pela OVX, mas não houve diferença nos níveis séricos de osteocalcina, expressão gênica ou relação vilo/cripta; indicando que a melhora nas propriedades biomecânicas não está associada à osteocalcina e que a perda óssea não se relacionou com alterações intestinais. L. reuteri, assim como a associação com MK-7, restauraram os valores de microarquitetura vertebral ao nível de SHAM, porém sem diferença para o grupo OVX. O crescimento microbiano in vitro foi avaliado associando-se L. reuteri à vitamina K2. Após os períodos de interações, foram realizadas espectrofotometrias das soluções, e as bactérias cultivadas em ágar MRS para contagem de unidades formadoras de colônias (UFC/ml). Houve aumento significativo da densidade óptica e de células viáveis após 4h de interação. L. reuteri e a associação com MK-7 melhoraram as propriedades biomecânicas de fêmures e mostraram tendência positiva na microarquitetura vertebral. O benefício da associação pode possivelmente estar relacionado à maior quantidade de L. reuteri que alcançou o intestino, por utilizar a vitamina para seu crescimento.
Osteoporosis is an osteometabolic disease, characterized by an imbalance between bone deposition and resorption. Probiotics and vitamin K2 (MK-7) have received attention as therapies for osteoporosis, independently. It is interesting to consider the association of probiotics with vitamin K, as their beneficial pathways could be interconnected, potentially resulting in synergistic effects. In this study, the effects of Limosilactobacillus reuteri associated with vitamin K2 (MK-7) on ovariectomy-induced bone loss in mice and bacterial growth in vitro were evaluated. In the in vivo study, animals were divided into five groups: sham-operated (control); ovariectomy (OVX); OVX + L. reuteri (LR); OVX + vitamin K2 (K); OVX + LR + K. After 4 weeks of treatment, the biomechanical properties of the femur, vertebral microarchitecture (L5), levels of carboxylated and non-carboxylated osteocalcin, gene expression of Occludin, Jam3, and intestinal villi and crypt heights were evaluated. Treatment with L. reuteri, MK-7, as well as their combination, significantly improved the biomechanical properties of the femur, which were deteriorated by OVX, but there was no difference in serum levels of osteocalcin, gene expression, or villi/crypt ratio, indicating that the improvement in biomechanical properties is not associated with osteocalcin and that bone loss is not related to intestinal changes. L. reuteri, as well as the combination with MK-7, restored vertebral microarchitecture values to the SHAM level, but without difference for the OVX group. Microbial growth in vitro was evaluated by combining L. reuteri with vitamin K2. After the interaction periods, optical density of the solutions was spectrophotometrically measured, and bacteria were cultured on MRS agar for colony-forming unit (CFU/ml) counting. There was a significant increase in optical density and viable cells after 4 hours of interaction. L. reuteri and the combination with MK-7 improved the biomechanical properties of femurs and showed a positive trend in vertebral microarchitecture. The benefit of the combination may be related to the greater amount of L. reuteri that reached the intestine by using the vitamin for its growth.
Osteoporosis is an osteometabolic disease, characterized by an imbalance between bone deposition and resorption. Probiotics and vitamin K2 (MK-7) have received attention as therapies for osteoporosis, independently. It is interesting to consider the association of probiotics with vitamin K, as their beneficial pathways could be interconnected, potentially resulting in synergistic effects. In this study, the effects of Limosilactobacillus reuteri associated with vitamin K2 (MK-7) on ovariectomy-induced bone loss in mice and bacterial growth in vitro were evaluated. In the in vivo study, animals were divided into five groups: sham-operated (control); ovariectomy (OVX); OVX + L. reuteri (LR); OVX + vitamin K2 (K); OVX + LR + K. After 4 weeks of treatment, the biomechanical properties of the femur, vertebral microarchitecture (L5), levels of carboxylated and non-carboxylated osteocalcin, gene expression of Occludin, Jam3, and intestinal villi and crypt heights were evaluated. Treatment with L. reuteri, MK-7, as well as their combination, significantly improved the biomechanical properties of the femur, which were deteriorated by OVX, but there was no difference in serum levels of osteocalcin, gene expression, or villi/crypt ratio, indicating that the improvement in biomechanical properties is not associated with osteocalcin and that bone loss is not related to intestinal changes. L. reuteri, as well as the combination with MK-7, restored vertebral microarchitecture values to the SHAM level, but without difference for the OVX group. Microbial growth in vitro was evaluated by combining L. reuteri with vitamin K2. After the interaction periods, optical density of the solutions was spectrophotometrically measured, and bacteria were cultured on MRS agar for colony-forming unit (CFU/ml) counting. There was a significant increase in optical density and viable cells after 4 hours of interaction. L. reuteri and the combination with MK-7 improved the biomechanical properties of femurs and showed a positive trend in vertebral microarchitecture. The benefit of the combination may be related to the greater amount of L. reuteri that reached the intestine by using the vitamin for its growth.
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Palavras-chave
Osteoporose, Osteopenia, Limosilactobacillus reuteri, Vitamina K, Osteoporosis, Vitamin K