El objetivo principal del trabajo fue determinar la expresión de distintas citoquinas proinflamatorias por los leucocitos circulantes de alpaca, al enfrentamiento antigénico de extracto de macroquistes de Sarcocystisaucheniae a diversas dosis y tiempos de exposición. Los leucocitos en una concentración de 500 000 cel/ml fueron expuestos a concentraciones de 0.5, 1, 50, 500 y 1000 ng de extracto de macroquistes de Sarcocystisaucheniae e incubados en placas de cultivo por 1, 12 y 24 horas. Posteriormente se extrajo los ARN mensajeros (ARNm) totales de cada tratamiento utilizando Trizol, que se utilizaron para realizar la RT-PCR tiempo real empleando cebadores específicos de Interleuquinas (ILs): IL-1α y β, IL-6 y TNFα. Finalmente, se observó que los niveles de ARNm de IL 1α y TNFα generados a la hora eran detectables y comparativamente elevados con respecto al calibrador (leucocitos no expuestos a extracto).La IL 1β se encontró incrementada en la concentración de 1 ng/ml a 24 horas, mostrandouna cinética de expresión negativa en comparación con el control no tratado. IL-6 no se evidenció mediante RT-PCR tiempo real. Además se realizó la observación de viabilidad leucocitaria a los tiempos de 1, 12 y 24 horas permitiendo corroborar el efecto tóxico del extracto de macroquistes de S. aucheniae a altas concentraciones. Se concluye que el extracto de Sarcocystisaucheniae induce una respuesta inflamatoria tóxica sobre los leucocitos de alpaca con expresión de las interleuquinas detectadas.
Palabras clave:Sarcocystisaucheniae,Interleuquinas, pro inflamatorias, alpaca / --- The main aim of this research was to determine the expression of several leukocytes proinflammatory cytokines from alpaca to the antigenic confrontation of Sarcocystisaucheniaemacrocysts extracts at various doses and different exposure times. Leukocytes in a concentration of 500 000 cells / ml were exposed to concentrations of 0.5, 1, 50, 500 and 1000 ng of Sarcocystisaucheniaemacrocysts extracts and were incubated in culture dishes for 1, 12 and 24 hours. The total messenger RNA (mRNA) were extracted for each treatment using Trizol, which were used to perform real-time RT-PCR using specific primers for interleukins (ILs), IL-1α and β, IL-6 and TNF-α. It was observed that the generated mRNAlevelsof IL-1α and TNF-α in the first hour were detectable and higher in comparison to the calibrator (leukocytes were not exposed to extract). IL-1β was found increased in a concentration of 1 ng / ml at 24 hours, what it means that there is a negative kinetic expression compared with the untreated control group. IL-6 was not evidenced by RT-PCR real time. In addition, the observation about the leukocytes viability times at 1, 12 and 24 hours corroborated the toxic effect of the macrocysts extract of S. aucheniae at high concentrations. It is concluded that the extract of Sarcocystisaucheniae induces an inflammatory response with expression of detected interleukins.
Keywords: Sarcocystisaucheniae, interleukins, pro-inflammatory, alpaca
| Identifer | oai:union.ndltd.org:Cybertesis/oai:cybertesis.unmsm.edu.pe:cybertesis/1526 |
| Date | January 2012 |
| Creators | Hinostroza Solano, Rocío Luz |
| Publisher | Universidad Nacional Mayor de San Marcos |
| Source Sets | Universidad Nacional Mayor de San Marcos - SISBIB PERU |
| Language | Spanish |
| Detected Language | English |
| Type | info:eu-repo/semantics/bacherlorThesis |
| Source | Universidad Nacional Mayor de San Marcos, Repositorio de Tesis - UNMSM |
| Rights | info:eu-repo/semantics/openAccess |
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