Comparison of BASP1 and MARCKS protein exchange traits in the kidneys of rats with spontaneous

Исследовали возможные различия в обмене белков BASP1 и MARCKS в почках крыс со спонтанной гипертензией (линияSHR), путем сравнения уровней экспрессии мРНК этих белков в различных структурах почек. Работа выполнена на самцах крыс линии SHR (n=8) и крыс линии WKY (n=8), в возрасте 90 дней. Исследование проводили на образцах ткани из коркового и мозгового слоев почек. Уровни экспрессии мРНК BASP1 и MARCKS определяли методом ПЦР в реальном времени. Обнаружены достоверные различия (p<0.001) уровней экспрессии мРНК BASP1 и MARCKS между корковыми и мозговыми слоями почек у крыс линий SHR и WKY. При этом межлинейные различия в уровнях экспрессии мРНК обоих белков практически отсутствовали (p=0.050). Показано, что неравномерное распределение уровней экспрессии мРНКбелков BASP1 и MARCKS в мозговом и корковом слоях почек не зависит от уровня артериального давления и наличия спонтанной гипертензии This study aims to detect differences in BASP1 and MARCKS protein metabolisms in kidneys of spontaneously hypertensive rats by comparing their mRNA expression in different kidney structures. The work was performed on 90 days old male SHR (n = 8) and WKY (n = 8) rats. The study was carried out on tissue samples from cortical and medullar kidney layers. BASP1 and MARCKS mRNA expression levels were determined by real-time PCR. Significant differences (p <0.001) in the expression levels of BASP1 and MARCKS mRNA between cortical and medullar kidney layers in both SHR and WKY strain rats were found. Moreover, interlinear differences in the levels of both proteins’ mRNA expression were practically absent (p = 0.050). It was shown that uneven distribution of mRNA expression levels of BASP1 and MARCKS mRNA in kidneys medullar and cortical layers does not depend on the level of blood pressure and the presence of spontaneous hypertension.

4379 MARCKS protein is altered in naturally occurring model of asthma in horses

OBJECTIVES/GOALS: Asthma is a significant health concern that affects people of all ages worldwide. EAS demonstrates many of the pathophysiological characteristics of nonatopic human asthma, which has led EAS to be used as naturally occurring model. Previous work from our lab determined that MARCKS (Myristoylated Alanine Rich C Kinase Substrate) protein is an essential regulator of cellular inflammatory functions. In the current study, we hypothesized that MARCKS levels would be increased in BAL cell lysates from horses with EAS, and that inhibition of MARCKS in zymosan-stimulated BAL cells (ex vivo) would diminish respiratory burst. METHODS/STUDY POPULATION: Lysates were prepared from BAL cells isolated from horses with no, mild/moderate and severe EAS. Relative MARCKS protein levels were determined using equine specific MARCKS ELISA (MyBioSource). Cultured BAL cells were pretreated with a MARCKS inhibitor peptide (MANS), control peptide (RNS) or vehicle control and stimulated with zymosan for 5 hours. Reactive oxygen species levels were determined by luminescence to evaluate respiratory burst. Data were analyzed by One-way ANOVA (p<0.05). RESULTS/ANTICIPATED RESULTS: We determined that normalized MARCKS protein expression is significantly increased in BAL cell lysates from horses with mild/moderate or severe EAS, compared to horses with normal BAL cytology. Preliminary findings also suggest that MANS treatment of zymosan-stimulated equine BAL cells ex vivo attenuates levels respiratory burst. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings point to a possible role for MARCKS protein in the pathophysiology of EAS and support MARCKS inhibition as a potential therapeutic strategy.