Leaves, Red Beans Blueberry Extract Antioxidant Capacity And Apoptosis,

Looking for the new active ingredient in the plant,and screening physiologically active substances as leading compounds of synthetic drugs or target compounds,are one of the most concemed issues in many fields of bioactive substances research. Bamboo leaves are rich in flavonoids.Compared with the common flavonoids, bamboo leaves flavonoids possess the characteristics of more stable structure,and strong hydrophilicity.It can easily be uptaken into depth focus and exert direct effects, and is conducive to the development of health food and medicine.Most blueberry plants contain anthocyanin of active ingredient.Anthocyanin is more effective in scavenging oxygen free radicals than vitamins,in particular reactive oxygen species. Free radicals can be generated under many environmental fators and trigger a variety of aging and related diseases in human body,which can be prevented from the removal of free radicals.In Vitro studies can be used as a major means to study the biological activity of the various functional components extracts.Cell proliferation assay may reflect the basic biological toxicity of studies objects.The detection of free radical scavenging capacity may indicate the antioxidative ability of them.The detection of the rate of apoptosis may show the impact of the studied substance on apoptosis phenomenon.Research objectiveIn this paper,extracts of bamboo leaves and Lingonberry extract has been selected to explore their effect on A549 cell proliferation in vitro,and antioxidant capacity,and the inducement of the apoptosis.To analyze and compare the differences of anti-DNA injury,scavenging free radicals,as well as the role of anti-apoptosis between bamboo leaves extract and Lingonberry extract will provide a scientific basis for the further study and development of natural extracts.Research methods1.MTT assay was used to detect the cytotoxicity of the two extracts and the protective effect of cell proliferation; 2.DPPH free radical scavenging test and ultra-weak chemiluminescence assay were applied to measure the antioxidant capacity of the two extracts;3.The rate of apoptosis was measured by Flow cytometry;4.Western blot method was used to detect the expression of NF-kB in A549 cellResearch results1.The impact of natural extracts on the cell proliferation(1)No significant effect of bamboo leaves extract on cell survival was observed during range of 2.5～40mg / L in A549 cells(p＞0.05).At the concentration of 80mg/L,the cell viability was decreased to 73.99%,which was significantly different, compared with the control group(p＜0.05).(2) Within the concentration of 2.5～5mg/L,added Lingonberry extract stimulated A549 cell proliferation.At the concentration of 10mg/L,the cell viability was not significantly changed,compared with control(p＞0.05).Within the concentration of 20-80mg/L,the cell viability were statistically significant different compared with the control group(p＜0.05).Cell viabilities were decreased with the extract concentration increase,with a dose-response relationship(r = 0.96,p＜0.05). At the concentration of 80mg/L,the cell viability was decreased to 59.31%.(3) Within the concentration of 20～80mg/L,under the same concentration,the cell viability of Lingonberry extract was lower then that of the extract of bamboo leaves(p＜0.05 or p＜0.01),that is,the cytotoxicity of Lingonberry extract was higher than that of the extract of bamboo leaves.(4) When the two natural extracts was add into the PM_2.5-induced A549 cell injury model,low concentrations had no significant impact on the cell viability,but at high concentrations of PM_2.5 group(400～800 mg / L),the two extracts could reduce the cytotoxicity.The cell proliferation rate was 8%at PM_2.5 800mg / L.It was increased to 25.83%when bamboo leaves extract was added and increased to 35.38% when Lingonberry extract was added.2.Antioxidant capacity of the extracts(1) Natural extracts to remove the ability of DPPH radical Lingonberry extract and bamboo leaves extract had the abilities of removing DPPH free radicals,the IC₅₀ of them were 3.79mg/L and 7.02mg/L,respectively.The free radical scavenging capacity of Lingonberry extract was higher than one of the bamboo leaves extract,but compared with the ascorbic acid(IC₅₀ 2.43mg/L),both them were lower.(2) The ability of the natural extracts against hydroxyl radical-induced oxidative DNA damage.With the CuSO4-Phen-Vc-H₂O₂-DNA Determination of ultra-weak chemiluminescence for 960s,Lingonberry could significantly reduce the DNA damage peak value and delay the emergence of damage peaks,and the bamboo leaves extract showed ability for the delay of the appearance of DNA damage.Lingonberry extract showed stronger effect,compared with bamboo leaves extract at this time. When measurement time was prolonged,it was found that the emergence of the DNA damage peak was significantly delayed and the value of DNA damage peak was decreased with the increase of bamboo leaves extract concentration.3.Effect of the atural extracts on the rate of apoptosis(1) The impact of natural extracts on apoptosis rate At the concentration of 10～40mg/L,bamboo leaves extract had no significant effect on the A549 apoptosis rate in the early and late stage(p＞0.05),Lingonberry extract at the concentration of 20 mg/L could induce significant increase of A549 apoptosis rate in early stage(p＜0.01),at the concentration of 40mg/L,it could not only increased the apoptosis rate in early stage,but also induced a marked increase in late stage(p＜0.01),and ncreased the degree of mechanical damage(p＜0.01).(2)Effect of Bamboo leaves extract on the PM2.5-induced A549 apoptosis The added PM_2.5(100mg /L) as injure model,selected the concentration of of as a model,add bamboo extract,its late stage apoptosis rate and mechanical injury rates were significantly lower(p＜0.01).However,changes in the rate of early stage apoptosis did not decrease significantly,not only not diminished but increased,the difference was statistically significant(p＜0.01).4.The relativity of PM_2.5 induced increases of expression of NF-kB in A549 cell and the appearance of bamboo leaves extract.100mg/L of PM2.5 can cause the high expression of NF-kB in A549 nucleus,there is statistically significant difference(p＜0.05).The appearance of 40mg/L of bamboo leaves extract has the role of a significant reduction at the high expression(p＜0.05).Conclusion1.The cytotoxicity of the extracts were both relatively low,by the PM_2.5-induced condition,the changes in cell proliferation can be against by bamboo extract and Lingonberry extract.The former is stronger.2.Lingonberry's ability to clear the DPPH free radical is stronger than bamboo extract, but both lower than that of ascorbic acid;two extracts have the role of anti-DNA damage,Lingonberry extract was stronger than bamboo extract.3.A certain concentration range of bamboo leaves extract has no significant influence on the apoptosis of A549 cell,but it aginst the increase of the late stage apoptosis of PM2.5-induced apoptosis on A549 cells;Lingonberry extract has the potential to induced apoptosis in A549 cells.4.A certain concentration of PM_2.5 could cause a significantly high expression of NF-kB in A549 nuclear,the concentration of the bamboo leaves extract could reduce the high expression significantly.