Beryllium Sulfate Induce Human Embryos Lung Apoptosis And The Preliminary Research Of The Lycium Barbarum Polysaccharides Protect Mechanism

Objective：In order to observe Sulfuric acid beryllium （BeSO₄） cause human embryoniclung fibroblast （MRC-5） reactive oxygen species （ROS） change; the expression level of apoptosisrelated gene c-fos BCL-2bax Caspase-3and changes of inner cell calcium concentration and3phosphoric acid inositol receptor （IP3R） expression, we using in vitro methods to discusseswhether sulfuric acid beryllium oxygen free radical influence mechanism through cell apoptosis,and observe LBP （LBP） whether to send a person to sulfuric acid beryllium embryo lung cellsdamage protective.Methods:Human embryonic lung fibroblast of in vitro culture have six groups: including blank group,low, medium and high sulfuric acid beryllium groups, LBP protection group, and LBP controlgroup. Sulfuric acid beryllium doses were given for0,1,10, and100μmol/L. LBP dose was10μmol/L. When the growth of cells to achieve the cell density logarithm growth period, add the24hdrug cultivation collects cells, the experiment goes on according to the following programs:1.The cytometry detection cells reactive oxygen species （ROS）;2. Immunohistochemical （SP）method to detect apoptosis related gene: c-fos, bcl-2, bax, Caspase-3expression.3. Confocallaser method to detect cells [Ca2+] level;4. Expressions of IP3R mRNA were determined withRT-PCR. Result:1. As the increase of sulfuric acid beryllium doses, express cell number increase, there weresignificant differences in following two groups: sulfuric acid active oxygen content and blankberyllium group was higher than the control group （P <0.05）.2. Cell dosing training after24h, Immunohistochemical results show: The express levels ofc-fos and bcl-2in low, medium and high BeSO₄dose groups were higher than the control group,with the drug dosage increased expression level increased and all have significantdifferences（P<0.05）; The express levels of bax in medium and high BeSO₄dose groups werelower than that of the control group, the express levels of Caspase-3in low,medium and highdose groups were lower than that of the control group, and with the drug dose increase theexpression level will be reduced,the difference was statistically significant （P <0.05）.3. Ca2+testing results: there were significant differences in sulfuric acid berylliumhigh-dosage groups and the control group.Compared with blank in cells group, Sulfuric acidberyllium high-dosage groups had a higher Ca2+concentrations.4. In the confocal laser under the microscope,100μmol/L BeSO₄group, the nucleus presenta bright green fluorescent, a clear demarcation between cells with a spindle type, BeSO₄+LBPgroup of cells are round, cells of the light green, green fluorescence visible to surrounding cellslittle point in cells.5. There were significant differences between IP3Râ…¢ in low, medium and high BeSO₄dosegroups and the control group （P<0.01）.6. BeSO₄damage the cells to LBP protection: ROS content of BeSO₄group was higher thanthe control group （P <0.01）.ROS content of LBP group was lower than the control group （P <0.01）, and both have interaction, had statistically significant （P <0.05）. BeSO₄and LBP groupshad the interactive effect in the expression of BCL-2and Caspase-3（P <0.05; P <0.01）.Ca2+and IP3R tests showed that BeSO₄and LBP both had interaction （P <0.05; P <0.01）. Conclusion:1. BeSO₄can cause MRC-5cell ROS content increased, LBP may reduce the cell by ROScontent on the produce protection.2. BeSO₄poisonous MRC-5cells were present c-fos, bcl-2gene high expression; while bax,Caspase-3low protein gene expression.3.Through the Ca2+receptor channels,BeSO₄may cause the increasing of MRC-5cellscalcium ion concentration.4.LBP may reduce the cell can adjust the IP3R type receptor expression regulation within thecell calcium ion concentration.