The new application of phenylephrine hydrochloride
A phenylephrine and pharmaceutical technology, applied in the field of medicine, can solve the problem of no phenylephrine hydrochloride and the like
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Embodiment 1
[0039] Embodiment 1, the in vitro test that phenylephrine hydrochloride (Phenylephrine Hcl) promotes rat cardiomyocyte proliferation
[0040] (1) Culture of Cardiomyocytes of SD Rats
[0041] Cardiomyocytes from SD rats born 3 days old were isolated and cultured in DMEM high glucose medium (Hyclone) + 5% horse serum (GIBCO) at 37 degrees in a 5% carbon dioxide incubator.
[0042] (2) Experimental grouping and processing
[0043] Isolate cardiomyocytes from SD rats, add 5% horse serum (GIBCO) + DMEM high-glucose medium (Hyclone), and add cytarabine (final concentration 20umol / L) to inhibit the growth of non-cardiomyocytes. Infect cTnT-mAG-hGeminin (1 / 110) virus (MOI value of virus infection=100) after 48 hours of adherence, after another 24 hours, replace with DMEM culture medium containing 0.5% FBS, add drugs in groups, and group as follows:
[0044] a. Experimental group: treated with phenylephrine hydrochloride (the final concentration in the medium is 2 μmol / L) for 24 hours...
Embodiment 2
[0050] Embodiment 2, the in vitro test that phenylephrine hydrochloride promotes neonatal rat cardiomyocytes to enter the cell cycle
[0051] Isolate the rat cardiomyocytes born 3 days old, add phenylephrine hydrochloride (final concentration in the culture medium is 2 μ mol / l), use DMEM+DMSO as the control (control), wash three times with PBS after 48 hours, 4% PFA ( Paraformaldehyde) fixed at room temperature for 15 min, washed three times with PBS, blocked at room temperature with 1% BSA / PBS / 0.1% Triton for 1 h, and added primary antibody after 1 h: Anti-α-Actinin (1:300, sigma, mouse, A7811), Phospho- Histone H3 (Ser10) antibody (1:300, CST, rabbit, 9701S), Anti-Ki67 (1:300, abcam, rabbit, ab15580) overnight at 4°C, washed 4 times with 1% BSA / PBS / 0.02% Tween, Add secondary antibodies: goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody (1:500, Invitrogen, A32731), goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody (1:500 , Life technologies...
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