Tamoxifen associated to the conservative CKD treatment promoted additional antifibrotic effects on experimental hypertensive nephrosclerosis.

CKD progression depends on the activation of an intricate set of hemodynamic and inflammatory mechanisms, promoting renal leukocyte infiltration, inflammation and fibrosis, leading to renal function loss. There are currently no specific drugs to detain renal fibrogenesis, which is a common end-point for different nephropathies. Clinical therapy for CKD is mostly based on the management of hypertension and proteinuria, partially achieved with renin-angiotensin-aldosterone system (RAAS) blockers, and the control of inflammation by immunosuppressive drugs. The aim of the present study was to verify if the administration of tamoxifen (TAM), an estrogen receptor modulator, clinically employed in the treatment of breast cancer and predicted to exert antifibrotic effects, would promote additional benefits when associated to a currently used therapeutic scheme for the conservative management of experimental CKD. Wistar rats underwent the NAME model of hypertensive nephrosclerosis, obtained by daily oral administration of a nitric oxide synthesis inhibitor, associated to dietary sodium overload. The therapeutic association of TAM to losartan (LOS), and mofetil mycophenolate (MMF) effectively reduced the severe hypertension, marked albuminuria and glomerular damage exhibited by NAME animals. Moreover, the association also succeeded in limiting renal inflammation in this model, and promoted further reduction of ECM interstitial accumulation and renal fibrosis, compared to the monotherapies. According to our results, the association of TAM to the currently used conservative treatment of CKD added significant antifibrotic effects both in vivo and in vitro, and may represent an alternative to slow the progression of chronic nephropathy.

Caffeine Accelerates Cystic Kidney Disease in a Pkd1-Deficient Mouse Model.

BACKGROUND/AIMS: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by progressive cyst formation and growth, leading to end-stage renal disease. A higher kidney volume is predictive of a more accelerated decline in renal function. This study aimed to examine the effects of caffeine, a phosphodiesterase inhibitor, on the progression of cystic kidney disease in a mouse model orthologous to human disease (Pkd1cond/cond:Nestincre). METHODS: Caffeine was administered to male cystic (CyCaf) and noncystic (NCCaf) mice (Pkd1cond/cond) from conception and at the postweaning period through 12 weeks of life (3 mg/d), while control animals consumed water (CyCtrl and NCCtrl). Renal ultrasonography was performed at 10 weeks of life to calculate total kidney volume and cystic index. At the end of the protocol, blood and urine samples were collected for biochemical analysis, and animals were euthanized. Kidneys were harvested to obtain renal tissue for determinations of adenosine 3´5´-cyclic monophosphate (cAMP) by an enzymatic immunoassay kit and phosphorylated extracellular signal-regulated kinase (p-ERK) by Western blotting. Renal fibrosis (picrosirius staining), renal cell proliferation (ki-67 immunohistochemistry) and apoptotic rates (TUNEL analysis) were also determined. RESULTS: At 12 weeks, CyCaf mice exhibited higher serum urea nitrogen, renal cystic index, total kidney volume, kidney cell proliferation, apoptosis and fibrosis compared with CyCtrl mice. Serum cystatin C was significantly higher in CyCaf than in NCCaf and NCCtrl mice. CyCaf mice had higher total kidney weight than all other groups but not higher heart and liver weight. The levels of cAMP and p-ERK did not differ among the groups. CONCLUSION: Caffeine consumption from conception through 12 weeks led to increased cystic index and total kidney volume and worsened renal function in Pkd1-deficient cystic mice, suggesting that high consumption of caffeine may contribute to a faster progression of renal disease in ADPKD.

Detecção de Mycoplasma pulmonis no trato respiratório superior em roedores através da técnica de PCR / /Mycoplasma pulmonis detection in the rodents respiratoru tract by PCR

Objetivo: apresentar a técnica de PCR para a detecção de Mycoplasma pulmonis das colônias convencionais de camundongos das linhagens isogênicas C57BL/6, BALB/c e heterogênica Swiss, e ratos Wistar. Material e Métodos: A extração do DNA foi realizada a partir de fragmento de traqueia, utilizando para a reação de PCR os oligonucleotídeos iniciadores F: 5’- CA TGT TCT TGG CCA CCTAT - 3’ e R:5’ - ATC CCT CAA TGA GCT GGT TC - 3' que amplificam um produto de 152 pb. Os produtos obtidos foram separados por eletrólise em gel de agarose. A visualização e fotodocumentação foram realizadas sob luz ultravioleta. Resultados e conclusões: A PCR possibilitou a detecção de camundongos positivos, sendo 38% da linhagem C57BL/6, 24% da linhagem BALB/c, 14% da linhagem heterogênica Swiss e 52% de ratos Wistar. Esses resultados sugerem que o emprego da técnica de PCR foi eficaz para o diagnóstico de Mycoplasma pulmonis em colônias de animais de laboratório. Certificado CEAU nº 251/11 HCFMUSP.

Objective: To introduce the PCR technique for detection of Mycoplasma pulmonis in conventional colony of mice inbred strains C5BL/6, BALB/c, outbred Swiss, and Whistar rats. Methods: DNA extraction was made from a fragment of the trachea, using the PCR primers F: 5’-CA TCT TGG TGT CCA CCTAT - 3’ and R:5’ - ATC CCT CAA TGA GCT GGT TC-3' which amplify a product of 152 bp. The products were separated by agarose gel electrophoresis. The viewing and photo documentation were performed under ultravioleta light. Results and Conclusion: The PCR allowed the detection of positive mice, 38% of C57BL/6, 24% of BALB/c 14% of Swiss and 52% of Wistar rats. These results suggest that the use of PCR technique was effective for the diagnosis of Mycoplasma pulmonis in colonies of laboratory animals. Certified CEAU nº 251/11 HCFMUSP.

Detecção de Mycoplasma pulmonis no trato respiratório superior em roedores através da técnica de PCR / /Mycoplasma pulmonis detection in the rodents respiratoru tract by PCR

Objetivo: apresentar a técnica de PCR para a detecção de Mycoplasma pulmonis das colônias convencionais de camundongos das linhagens isogênicas C57BL/6, BALB/c e heterogênica Swiss, e ratos Wistar. Material e Métodos: A extração do DNA foi realizada a partir de fragmento de traqueia, utilizando para a reação de PCR os oligonucleotídeos iniciadores F: 5- CA TGT TCT TGG CCA CCTAT - 3 e R:5 - ATC CCT CAA TGA GCT GGT TC - 3' que amplificam um produto de 152 pb. Os produtos obtidos foram separados por eletrólise em gel de agarose. A visualização e fotodocumentação foram realizadas sob luz ultravioleta. Resultados e conclusões: A PCR possibilitou a detecção de camundongos positivos, sendo 38% da linhagem C57BL/6, 24% da linhagem BALB/c, 14% da linhagem heterogênica Swiss e 52% de ratos Wistar. Esses resultados sugerem que o emprego da técnica de PCR foi eficaz para o diagnóstico de Mycoplasma pulmonis em colônias de animais de laboratório. Certificado CEAU nº 251/11 HCFMUSP. (AU)

Objective: To introduce the PCR technique for detection of Mycoplasma pulmonis in conventional colony of mice inbred strains C5BL/6, BALB/c, outbred Swiss, and Whistar rats. Methods: DNA extraction was made from a fragment of the trachea, using the PCR primers F: 5-CA TCT TGG TGT CCA CCTAT - 3 and R:5 - ATC CCT CAA TGA GCT GGT TC-3' which amplify a product of 152 bp. The products were separated by agarose gel electrophoresis. The viewing and photo documentation were performed under ultravioleta light. Results and Conclusion: The PCR allowed the detection of positive mice, 38% of C57BL/6, 24% of BALB/c 14% of Swiss and 52% of Wistar rats. These results suggest that the use of PCR technique was effective for the diagnosis of Mycoplasma pulmonis in colonies of laboratory animals. Certified CEAU nº 251/11 HCFMUSP.(AU)