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Influence of pine pollen on deletion mutation of human mitochondrial DNA in aging fibroblast cells

YU Lu,SHI Chun-Xia. Chinese PLA 305 Hospital,Beijing 100017,China  
Objective To detect the influence of pine pollen on the mitochondria DNA(mtDNA) deletions of aging cells and investigate the antidotal mechanism of pine pollen.Methods Cells were divided into three groups: the young cells were determined as group A and fed by normal DMEM,aging cells were determined as group B and fed by normal DMEM,aging cells in the research group fed by DMEM containing 240 mg/dl pine pollen and named as group C.The intracellular total mtDNA was extracted from the cells in different groups.Using a conservative fragment in the mtDNA as an internal standard,the relative proportion of mtDNA4977 in the total mtDNA was determined by a competitive polymerase chain reaction(PCR) method,the positive rate of SA-β-galactosidase stain was determined by immunohistochemistry.Relative level of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) in cells were measured.Results After the treatment of pine pollen,positive rate of SA-β-gal was significantly decreased(P0.05),pine pollen could down regulate the proportion of the mtDNA4977 deletion in the total mtDNA of the aging cells.Pine pollen could decrease MDA volume and increase the activity of SOD significantly(P0.05).Conclusions Pine pollen can reduce oxidative damage of mtDNA and protect mtDNA.Accordingly it provides the possible mechanism of anti-aging effect of Pine pollen from the molecular level.
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