Effect And Mechanism Of Zingiber Officinale On Cell Proliferation And Apoptosis Of Human Lung Adenocarcinoma 549

Objective To investigate the apoptosis effects of Zingiber officinale in human lung Adenocarcinoma 549 (A549) and the possibie mechanism of apoptosis.Methods The different dose (10μmol/L,20μmol/L,40μmol/L ) of Zingiber officinale were interacted with A549 cell line for 24h in the experimental group compared to contrast group.1. The multiplication capacity of A549 cells by Zingiber officinale was analyed using.2. By light microscope the growth and morphological changes of A549 cells were observed.3. By in situ terminally labeled transferase technique(TUNEL) apoptotic were detected.4. By SP immunocytochemical method the expression of Cyt C protein of A549 cells were detected.5. The mitochondria membrane potential were determined. By flow cytometry the flutorescence intensity of rhodamine 123 were determined,thus the mitochondria membrane potential can be assumed.6. By flow cytometry technique the apoptosis were detected.Results1. MTT assay showed that the inhibition rate of cell proliferation was 0.04% in control group, and 14.87%,24.72%,38.21% in experiment group, respectively. The inhibition rate was higher in experiment group compared to the control group significantlly(p<0.01); the inhibition rate increased as the concentration of the Zingiber officinale increased, and there were significant difference between each dose of Zingiber officinale (p <0.01).2. The light microscope showed few apoptotic cells in the control group. But there were scattered cells with more apoptotic cells in the experiment group and it`s much significant as the concentration of the Zingiber officinale increased, especially with a dose of 40μmol/L.3.The TUNNEL test showed the index of apoptotic ceslls was 4.81±0.36% in the control group compared to 9.36%±0.82%,13.74%±1.15%,19.17%±1.4% in the experiment group, The difference was significantly in both groups(p<0.01); it also significantly increased as the concentration of the Zingiber officinale increased.4. The SP immunocytochemical method showed level of xpression of Cyt C protein was 0.15±0.02 in the control group and 0.23±0.03,0.27±0.06,0.32±0.07 in the experiment group,it was significant difference between the two group(p <0.01). 5. The mitochondria fluorescence intensity on cells dyed by rhodamine 123 that was determined through flow cytometry technique were 16.41±2.32 in the control group and 24.78±3.88, 36.47±5.74, 47.89±6.92 in the experiment group, there`s significant difference between the two group(p <0.01).6. The flow cytometry technique showed the apoptosis rate was 0.17±0.03% in the control group and 9.87%±1.86%,12.54%±2.12%,18.39%±1.98% in the experiment group, there`s significant difference between the two group(p <0.01).Conclusion1. Zingiber officinale could inhibit the cell proliferation of A549 cell line and the effect was significantly dose-dependent, showed in this experiment.2. The mitochondrial membrane potential of the A549 cell apoptosis decreased significantly caused by the Zingiber officinale(p<0.01). The SP immunocytochemical method showed that the expression of Cyt C protein increased as the Zingiber officinale increased. Zingiber officinale could inhibit the multiplication capacity of the A549 cell and induce it`s apoptosis. The mechanisme may be the decreasing of the mitochondrial membrane potential that caused the apoptosis of the A549.