The Cytotoxicity Caused By Cigarette Smoke Condensate And Protection Of TPs

Objective:1,Discusses the cigarette smoke extraction inclusions (CSC) to bronchial epithelium cells (BEAS-2B) oxidative damage effect and its mechanism;2,Observe the tea polyphenol (TPs) to cigarette smoke cause person BEAS-2B cells oxidative damage of protection for its mechanism, cigarette smoke injury medical protection offers experiment basis.Methods:Using JJ-100 type of single channel smoking cigarette smoke machine will be absorbed in anhydrous alcohol and LHC-8 medium, mixed made after extraction things experiment, Trials are divided into eight groups:blank group, CSC low, medium and high concentrations of poisonous group, TPs control, TPs protection high, medium and low CSC concentration poisonous group,3 CSC low, medium and high concentrations of poisonous gas extraction groups will inclusions (CSC) according to design thick join BEAS-2B cells medium℃next incubation 2h; 37 TPs protection is low, medium and high concentrations of poisonous group, first adds 250 mg/L TPs buy BEAS-2B cells medium 37℃next incubation 2 hours, join CSC poisonous continue incubation 2 hours, Blank group just add BEAS-2B cells, TPs 250 mg/L just add L TPs 37℃next incubation 2 hours.Using the comet experiment and gamma-H2AX expression detection cell DNA damage, HPRT gene mutations detection point mutations, micro nuclear test detection genome damage, using stereology detection of cells to study for CSC malignant cells hereditary toxicity function and TPs protective effect, By spectrophotometry and fluorescence spectrophotometry measure cellular internal and external superoxide, hydrogen peroxide, hydroxyl radicals concentration, using chemiluminescence method to detect the change and SOD activity of enzyme LDH inside cells escape detection membrane injury.Results:1,Comets experiment shows that the CSC cause BEAS-2B cell DNA double chain rupture, causing cell bad-tail rate, comets tail DNA content, tail long, tails area, the tail from increased with increase blank group compared are statistically significant difference (P<0.05), TPs and group compared with doses of CSC DNA damage mitigation (P<0.05);2,WESTBLOT experiment shows that the CSC causes the cell gamma- H2AX protein expression increases, caused the DNA damage, TPs with doses of CSC group compared to reduce DNA damage;3.Micro nuclear test showed that CSC can cause BEAS-2B cells of the micro kernel rate increased, high dose group of micro kernel rate reaches as high as 79 per group compared with blank was statistically significant difference (P<0.05), TPs with doses of CSC group compared micro nuclear rate to decrease (P<0.05);4,HPRT mutation rate test indicated:CSC can cause HPRT mutation rate increases, the highest can achieve 1.058 per group compared with blank was statistically significant difference (P<0.05), TPs with doses of CSC group compared mutation rate reduce (P<0.05);5,Stereology analysis shows that the CSC can cause cells cell bodies and nuclei area, minimum diameter, equivalent diameter and form factor change, relative blank cells form tendency of malignant group (P<0.05), TPs for CSC cause cell type malignant change change are apparent inhibition (P<0.05);6,Cell ROS test indicated:CSC can lead to extracellular superoxide, hydrogen peroxide, hydroxyl radicals, inside the cell superoxide, hydrogen peroxide, have all increased doses-effect relation, with blank group compared are statistically significant difference (P<0.05), TPs with doses of CSC group compared ROS concentration lower (P<0.05);7,SOD and LDH activity test indicated:CSC can lead to cells, causing cell membrane lower SOD activity impaired, permeability increase, making the cell inside of overflow, and blank LDH group compared are statistically significant difference (P< 0.05), TPs with doses of CSC group compared damage reduced (P<0.05).Conclusion:1,CSC BEAS-2B by inducing cell produces ros, causing cell membranes, leading to oxidative damage cell membrane permeability increase, genetic material damage;2,250mg/L TPs can reduce the CSC oxidative damage and achieve a protective effect.