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1.
Int J Mol Sci ; 25(10)2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38791162

RESUMO

Early detection of drug-induced kidney injury is essential for drug development. In this study, multiple low-dose aristolochic acid (AA) and cisplatin (Cis) injections increased renal mRNA levels of inflammation, fibrosis, and renal tubule injury markers. We applied a serum amyloid A3 (Saa3) promoter-driven luciferase reporter (Saa3 promoter-luc mice) to these two tubulointerstitial nephritis models and performed in vivo bioluminescence imaging to monitor early renal pathologies. The bioluminescent signals from renal tissues with AA or CIS injections were stronger than those from normal kidney tissues obtained from normal mice. To verify whether the visualized bioluminescence signal was specifically generated by the injured kidney, we performed in vivo bioluminescence analysis after opening the stomachs of Saa3 promoter-luc mice, and the Saa3-mediated bioluminescent signal was specifically detected in the injured kidney. This study showed that Saa3 promoter activity is a potent non-invasive indicator for the early detection of drug-induced nephrotoxicity.


Assuntos
Ácidos Aristolóquicos , Luciferases , Regiões Promotoras Genéticas , Proteína Amiloide A Sérica , Animais , Proteína Amiloide A Sérica/genética , Proteína Amiloide A Sérica/metabolismo , Camundongos , Luciferases/metabolismo , Luciferases/genética , Ácidos Aristolóquicos/toxicidade , Genes Reporter , Cisplatino/toxicidade , Cisplatino/efeitos adversos , Medições Luminescentes/métodos , Masculino , Nefropatias/induzido quimicamente , Nefropatias/genética , Nefropatias/metabolismo , Nefropatias/patologia , Rim/metabolismo , Rim/efeitos dos fármacos , Rim/patologia , Modelos Animais de Doenças , Camundongos Endogâmicos C57BL
2.
Int J Mol Sci ; 23(2)2022 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-35055081

RESUMO

The early detection of diabetic nephropathy (DN) in mice is necessary for the development of drugs and functional foods. The purpose of this study was to identify genes that are significantly upregulated in the early stage of DN progression and develop a novel model to non-invasively monitor disease progression within living animals using in vivo imaging technology. Streptozotocin (STZ) treatment has been widely used as a DN model; however, it also exhibits direct cytotoxicity to the kidneys. As it is important to distinguish between DN-related and STZ-induced nephropathy, in this study, we compared renal responses induced by the diabetic milieu with two types of STZ models: multiple low-dose STZ injections with a high-fat diet and two moderate-dose STZ injections to induce DN. We found 221 genes whose expression was significantly altered during DN development in both models and identified serum amyloid A3 (Saa3) as a candidate gene. Next, we applied the Saa3 promoter-driven luciferase reporter (Saa3-promoter luc mice) to these two STZ models and performed in vivo bioluminescent imaging to monitor the progression of renal pathology. In this study, to further exclude the possibility that the in vivo bioluminescence signal is related to renal cytotoxicity by STZ treatment, we injected insulin into Saa3-promoter luc mice and showed that insulin treatment could downregulate renal inflammatory responses with a decreased signal intensity of in vivo bioluminescence imaging. These results strongly suggest that Saa3 promoter activity is a potent non-invasive indicator that can be used to monitor DN progression and explore therapeutic agents and functional foods.


Assuntos
Nefropatias Diabéticas/metabolismo , Expressão Gênica , Genes Reporter , Luciferases/genética , Imagem Molecular , Regiões Promotoras Genéticas , Proteína Amiloide A Sérica/genética , Animais , Biomarcadores , Diabetes Mellitus Experimental , Nefropatias Diabéticas/diagnóstico , Nefropatias Diabéticas/etiologia , Imunofluorescência , Regulação da Expressão Gênica , Mediadores da Inflamação/metabolismo , Medições Luminescentes/métodos , Camundongos , Imagem Molecular/métodos , Transcriptoma
3.
PLoS One ; 10(6): e0128002, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26061268

RESUMO

Label-free, non-invasive, rapid absorbance spectral imaging A(x,y,λ) microscopy of single live cells at 1.2 µm × 1.2 µm resolution with an NA = 0.85 objective was developed and applied to unicellular green algae Chlamydomonas reinhardtii. By introducing the fiber assembly to rearrange a two-dimensional image to the one-dimensional array to fit the slit of an imaging spectrograph equipped with a CCD detector, scan-free acquisition of three-dimensional information of A(x,y,λ) was realized. The space-resolved absorbance spectra of the eyespot, an orange organelle about 1 µm, were extracted from the green-color background in a chlorophyll-rich single live cell absorbance image. Characteristic absorbance change in the cell suspension after hydrogen photoproduction in C. reinhardtii was investigated to find a single 715-nm absorption peak was locally distributed within single cells. The formula to calculate the absorbance of cell suspensions from that of single cells was presented to obtain a quantitative, parameter-free agreement with the experiment. It is quantitatively shown that the average number of chlorophylls per cell is significantly underestimated when it is evaluated from the absorbance of the cell suspensions due to the package effect.


Assuntos
Clorófitas/química , Microespectrofotometria/métodos , Chlamydomonas reinhardtii/química , Clorofila/química , Suspensões/química
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