RESUMEN
PURPOSE: Proteomics is a peptide screening approach to identify proteins related to physiological and pathological changes. Our objective was to analyze changes in serum proteome profiles in patients suffering from septic shock, during the first 24-h period of therapy and to determine whether these changes were associated with prognosis. METHODS: This was a retrospective observational study based on prospectively collected data that was conducted in an emergency intensive care unit (ICU) of a tertiary referral hospital. Consecutive patients admitted to the ICU with septic shock were enrolled. We obtained serum samples from the patients at admission (0 h) and 24 h after admission (24 h). Then, we analyzed the serum proteome profiles, performed Western blots, and measured serum lipid profiles. RESULTS: Twenty-seven patients were enrolled. Thirteen patients were grouped into the survivors (SURV) group and fourteen into the non-survivors group on day 30 (NON-SURV). Proteomic analyses and Western blot showed that the expression intensities of proapolipoprotein AI remained unchanged in SURV but decreased in NON-SURV during the first 24-h period of septic shock (p=0.015). Serum high density lipoprotein (HDL) cholesterol level also remained unchanged in SURV but decreased in NON-SURV (p=0.036). CONCLUSION: The decrease in serum proapolipoprotein AI expression during the first 24-h period of therapy was associated with a consistently low serum HDL cholesterol level and a poor prognosis in patients with septic shock.
Asunto(s)
Humanos , Apolipoproteína A-I , Western Blotting , Colesterol , HDL-Colesterol , Urgencias Médicas , Unidades de Cuidados Intensivos , Lipoproteínas , Lipoproteínas HDL , Tamizaje Masivo , Pronóstico , Estudios Prospectivos , Proteínas , Proteoma , Proteómica , Estudios Retrospectivos , Choque , Choque Séptico , Estrés Psicológico , Sobrevivientes , Centros de Atención TerciariaRESUMEN
To facilitate the mutation of Milano site, the whole proapo AI_(M) gene was divided to two fragments to synthesize by RT-PCR. The optimization of upstream gene sequence was carried out to improve proapo AI_(M) expression level in E. coli. After induction with a shift of temperature, yields of recombinant proapo AI_(M) achieved about 45 % of total cell protein and the recombinant proapo AI_(M) was expressed as a form of inclusion body in cells. The renaturation of protein was carried with a hydrophobic interaction column,the results showed that recombinant proapo AI_(M) had similar functional properties identical to those of native protein.