Below 37uC, 5% CO2 and saturated ttigter humidity for cell growth If H9c2 cells reached 80% confluency 90, was changed to complete medium for serum-free medium for 24 h And then incubated in DMEM with 10% FBS. In the NaHS group each H9c2 cells with NaHS were at 100 mmol / l 30 min, min DM 100 mmol / l for 5, DTT 5 mmol / L for 5 min, the GSH min 5 mmol / l for the CY-treated 5 to 5 mmol / L for 5 minutes, and followed NaHS at 100 mmol / l of DTT for 25 min 5 mmol / l for 5 min with GSH, or 5 mmol / l for 5 minutes. While Volasertib BI6727 in the control group H9c2 cells  for the same period. H9c2 then cells were solubilized in 1 ml lysis buffer and cell lysates were incubated with 50 ml EZ LinkTM iodoacetyl biotin PEO 12 h at 4UC and then incubated with 30 ml of immobilized UltraLinkTM NeutrAvidinTM for 4 hours, a roller system 4UC. The beads were washed twice with 1 ml of lysis buffer and three times with 1 ml PBS.
For Western blot analysis, the proteins, the sulfhydryl groups of H9c2 cells were subjected to SDS-PAGE and the proteins Were transferred to nitrocellulose membranes. Membranes were incubated with antique Probed body against L-type calcium control and developed with Western blotting luminol. Statistical analysis Data were analyzed using the PClamp 10.0, 6.0 and SPSS 13.0 software Microcal Origin. All data in the figures are expressed as mean 6 SD. The differences between the groups. With ANOVA followed by LSD or Dunnett’s post hoc test, where appropriate Significance was at P, set 0.05. Results The effect of NaHS on cardiac function perfusion with 100 mmol / L NaHS at a dose continuous Salzl Solution for 10 min, 6 LV dp / dt max and decreased DLVP fa Is significant compared to control.
Sulfhydryl modifiers investigate effects NaHS-induced inhibition of cardiac function in isolated rat hearts whether perfused NaHS isolated an inhibitory effect on the heart function dependent perfused rat heart-Dependent sulfhydryl group of proteins induced, we used DM a modifier oxidation sulfhydryl transform the sulfhydryl groups of proteins in disulfide . The LV and 6dp/dtmax DLVP sank after perfusion with DM min at a dose of 100 mmol / L for 5 compared to the control group. However, the presence of fluid infusion DM, LV and 6dp/dtmax DLVP not ver Changed if they continuously perfused with 100 mmol / l NaHS 10 min. Then we DTT, k is a sulfhydryl modifying agent in the perfusion fluid, whether mediate induced inhibition of cardiac function by NaHS Nnte.
Zus Tzlich to the fact that BT 6DP / dt max and DLVP not w During perfusion with 100 mmol / l DTT, 5 min changed in comparison with the controls ver, We found there continuous infusion of L-KH solution with 100 mmol / L NaHS for 10 min in the presence of DTT decreased obviously 6dp/dtmax DLVP LV and, based on the untreated TNT infusion NaHS. The effect of nifedipine on cardiac function perfused isolated rat hearts compared to controls treated with NaHS, the LV and 6dp/dtmax DLVP reduced if with KH L Perfused solution of nifedipine assembled in a dose of 10 mmol / l for 5 minutes. But after a continuous infusion of L-KH Solution for 10 min, the ventricular erh Ren rate and 6dp/dtmax DLVP Fa ht You clearly to those with L KH Composed solution of nifedipine compared. Moreover, the data showed that a continuous infusion of NaHS in a dose of 100 mg / dl after infusion of the ventricular Re frequency and 6dp/dtmax DLVP erh hen.