ABSTRACT
Objective To study the effects of acetylcholine combined with vincristine (VCR) on A549 cells, furthermore to study the influence that is due to affecting the expression of the two proteins bcl-2 and p53. Methods MTT assay was used to analyze growth inhibition effect of acetylcholine alone and combined with IC50VCR on A549 cells.The expression of bcl-2 and p53 was measured by immunohistological chemistry technique(IHC) and Western blot.Results The inhibition rates for A549 cells with 0.1,1 and 10 μmol/L acetylcholine were (0.050±0.032)％,(0.101±0.021)％,(0.169±0.015)％.The inhibition rates for tumor cells with 0.1,1 and 10 μmol/L acetylcholine joint 0.2 μg/ml VCR were (0.529±0.023)％,(0.545±0.011)％,(0.589±0.015)％,the difference was statistically significant compared with the VCR group (q'values were 1.09,1.37,1.83,P＜0.05).Acetylcholine alone exerted inhibitory effect on A549 cells in a concentration dependent manner and significanrtly enhanced its sensitivity to VCR (P＜0.05).acetylcholibne (0.1-10 μmol/L)combined with IC50VCR decrased the expression of bcl-2 and p53 (P＜0.05).Conclusion Acetylcholine alone and combined with VCR can inhibit the growth of A549 cells. Significant synergistic effect between acetylcholine and VCR is found in inducing cell apoptosis by changing the expression of bcl-2 and p53.
ABSTRACT
Objective To explore the correlation of atheroselerosis progression and the expression of platelet derived endothelial nitric oxide synthase (eNOS) in rabbits. Methods A total of 24 male New Zealand white rabbits were used in this study. Six of the animals were fed with normal food (control group). Eighteen rabbits were fed with cholesterol-rich food (1 g/d) for 12 weeks to establish the atherosclerosis model. Among 18 models, 6 rabbits were executed immediately and their aorta and platelet samples were collected for further analysis (model group), 6 rabbits were orally administered with pravastatin (10 rag/d) for additional 12 weeks (treated group), and the remaining 6 rabbits were left untreated until the end of the study (untreated group). The control, treated and untreated animals were then killed, and the aorta and platelet samples were collected for eNOS expression analysis (RT-PCR). Results The aorta samples in model and untreated group exhibited rough intima and a lot of longitudinal fatty streaks, which indicated that atherosclerosis models were established successfully. While in treated group, the degree of atherosclerosis was decreased. The average percent of thickness of fatty streaks or atheroselerotic plaques relative to the whole thickness of vessel walls was 0. 04±0. 02, 0. 82±0. 16, 0. 33±0. 18,0. 77±0. 14 in control, model, treated and untreated group, respectively. The thickness of fatty streaks or atherosclerotic plaques was significantly increased in the model and untreated groups and decreased in treated group compared with the control group (both P<0. 05). The expressions of platelet derived eNOS/mRNA were 1. 02± 0. 28, 0. 41± 0. 27, 1.00 ± 0. 77, 0. 40±0. 29 in control, model, treated and untreated group, respectively. The expression of eNOS/mRNA was markedly decreased in model group and untreated group compared with the control group, but was increased in treated group compared with untreated and model groups (F=3. 544, P = 0. 024). Conclusions There is a negative correlation between eNOS expression and atherosclerosis development, which suggests that the reversal effect of pravastatin on atheroselerosis progression and plaque formation may relate to the expression of platelet derived eNOS.