Anti-Aging Effect Of Extract From Maize Plumule On Drosophila Melanogaster And Testis In Mice

ObjectiveThis study was designed to investigate the effect of Extract from Maize Plumule on life-span, antioxydative ability of Drosophila melanogaster , the effect of testis ultrastructrue and function and antioxydative enzyme activity of aging mice by D-gal, the effect of testosterone synthesis and secretion of Leydig cells in primary culture from rats testis.Methods1.Establishment of aging model of Drosophila melanogaster. Collect eclosion imago in 8 h and divided into different groups. Life-span team: feed with different concentration medium(0 mg/kg, 20 mg/kg, 40 mg/kg, 80 mg/kg),then count the number of the survival Drosophila Melanogaster in every group everyday until the last one died, then work out the mean lifetime and maximum lifetime. SOD and MDA team: feed with D-gal and different concentration medium(0 mg/kg, 20 mg/kg, 40 mg/kg, 80 mg/kg), then test SOD activity in every group and MDA content on 10 d,20 d,30 d,40 d, every group. D-gal substitutes sucrose in normal medium, D-gal concentration is 6.5%, with sucrose compensate. Water content research: the fruitflies in 4 days were put into baking oven(55℃) for 1 hour, then got the content. Starvation: the fruitflies in 4 days were put into baking oven(25℃), then count the number of living fruitflies, three times everyday, until the last fruitflies died. Heat stress: the fruitflies in 10 days were put into baking oven(40℃), then count the number of living fruitflies, every 1 hour, until the last fruitflies died.2.Establishment of aging mice model caused by D-gal. Sub-acutely aging mice model was made by subcutaneous injection(sc.) with D-gal for six weeks continually, while the mice were treated trough injecting Extract from Maize Plumule into their gaster with three different concentration: 10 mg/kg, 20 mg/kg, 40 mg/kg. After six weeks, measured SOD, GSH-PX, NO, MDA and serum T, then observed the testis tissue changes under the light microscope ,and the ultrastructural changes of testis under transmission electron microscope.Then test sperm function in epididymis, including sperm number and motility rate.3.Establishment of aging Leydig cells model caused by H₂O₂. We got cells suspension from SD rats (250～300 g) testis with enzymatic digestion. Leydig cells were being cultured in DMEM/F12 with different concentration Extract from Maize Plumule for 18 hours. Then H₂O₂(30 %H₂O₂,50μM)was added to the medium to make cell aging model. After 6 hours, collected the medium to detect T and SOD and MDA. Leydig cells stained with haematoxylin- eosin. Survival rate of Leydig cells was tested by MTT test.Results1.Anti-aging effect of Extract from Maize Plumule on Drosophila Melanogaster.Extract from Maize Plumule(80 mg/kg) had the ability of longer anverage lifetime and longest lifetime of Drosophila Melanogaster, and increasing SOD activity(P<0.01)in 30 and 40 days, and reducing MDA content in 30 and 40 days(0.01< P<0.05), and increasing ability of anti-heatstress.2.Extract from Maize Plumule could increase SOD activity, GSH-PX activity, NO content, serum T content, and decrease MDA content remarkably, and increase sperm number and motility rate, specially group of 10 mg/kg. The ultrastructrue of testis of the mice with Extract from Maize Plumule changed obvirously.3.3β-HSD identifed primary cultured cells as Leydig cells. The group in 20 mg/L could help protect primary cultured Leydig cells from H₂O₂(30 % H₂O₂, 50μM), and maintained Leydig cells integrality in morphology, and increased the testosterone synthesis and secretion ability of Leydig cells from rat testis.Conclusions1.Extract from Maize Plumule have the effect of increasing lifetime and anti-oxidation ability on Drosophila melanogaster.2.Extract from Maize Plumule could increase anti-oxidation ability of testis, increase serum T level, and protect the ultrastructrue of aging mice testis caused by D-gal. Therefore Extract from Maize Plumule have the effect of anti-aging on testis.3.Extract from Maize Plumule could increase increase the testosterone synthesis and secretion ability for testis Leydig cells in primary culture.