Proteomic analysis of left ventricular diastolic dysfunction hearts in renovascular hypertensive rats.

Abnormalities of diastolic function are common to virtually all forms of cardiac failure. However, the molecular events leading to diastolic dysfunction have not been fully elucidated. We performed a differential proteomic profiling study on diastolic dysfunction hearts induced by renovascular hypertension. Left ventricular diastolic dysfunction induced by renovascular hypertension (2K1C, two-kidneys, one clip) was performed in twelve Sprague-Dawley rats. 2D echocardiographic and cardiac protein patterns (2D-electrophoresis and mass spectroscopy) were compared with the sham operated rats. We described sixteen altered protein spots in 2K1C rats with left ventricular diastolic dysfunction. Calsarcin-1 (CS-1) was significantly down-regulated in 2K1C rats and it showed a negative correlation with calcineurin enzymatic activity (r(2)=0.72 p=0.03). We also showed changes in cellular energy metabolism in 2K1C rats, and these changes go in parallel with alterations of the thin filament proteome responsible for actin-myosin cross-bridge. In conclusion, this study provides a new insight into the left ventricular proteome profile associated with systemic hypertension induced diastolic dysfunction in a renovascular hypertension rat model. The decreased CS-1 protein with a concomitant increased enzymatic activity of calcineurin, suggests an important role of CS-1 in the calcineurin-mediated left ventricular hypertrophy.

Construction of a recombinant adenovirus vector of the rat Lys184del mutant cardiac troponin I gene / 医学研究生学报

Objective: To construct a recombinant adenovirus expression vector of the SD rat Lys184del mutant cardiac troponin I gene(cTnI Lys184del).Methods: The Lys184del mutant cardiac troponin I gene of SD rat was induced by PCR-based site-directed mutagenesis,the amplified fragment containing the mutation was then inserted into the cloning vector pMD18-T.After the addition of a sequence encoding a 6?His tag at the C-terminus by PCR,the coding region of cTnI Lys184del was subcloned into Adeno-X viral DNA to form a recombinant adenoviral plasmid,which was packed into infectious recombinant adenoviral particles(Ad-cTnI Lys184del) by transfecting HEK293 cells.Results: The DNA sequence of Ad-cTnI Lys184del was proved to be correct by PCR and sequencing,and its expression was identified by Western blot analysis.Conclusion: The successfully constructed recombinant adenovirus Ad-cTnI Lys184del has provided valuable data for further researches on its functional characteristics.