[Inhibitory action of Potentilla anserine polysaccharide fraction on H2O2-induced apoptosis of murine splenic lymphocytes].
A water-soluble polysaccharide fraction from root of Potentilla anserine was obtained. Gas chromatogram, FT-IR, physical and chemical characteristics of the Potentilla anserine polysaccharide fraction (PAPF) were analyzed. The protective effects of PAPF against the H2O2 induced process of apoptosis of murine splenic lymphocytes were investigated in vitro. Morphological assessment of apoptosis was performed with light microscope and laser scanning confocal microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. The results showed that PAPF is composed of rhamnose, arabinose glucose and galactose. H2O2 (200 micromol x L(-1)) induced apoptosis of murine splenic lymphocytes with the cell volume reduced, cytoplasm and nuclear shrunk and DNA stained non-uniformly. Condensed chromatin and formation of apoptotic body were observed in the apoptotic cells. Apoptotic bodies in the cells treated with PAPF and H2O2 were less than those in H2O2 treatment alone. DNA fragmentation assay showed that PAPF (50, 100, 200, and 400 microg x mL(-1)) obviously reduced H2O2-induced ladder bands. Flow cytometry analysis showed that H2O2 increased the populations of apoptotic sub-G1 cells from 5.60% (control) to 45.40%, and PAPF decreased H2O2-induced apoptosis to 37.80%, 22.70%, 17.70%, and 8.50%, respectively. In conclusion, PAPF reduced H2O2-induced oxidative damage in a dose dependent manner.
Shuai
Xue-Hong
XH
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China.
Hu
Ting-Jun
TJ
Zhang
Xia
X
Cheng
Fu-Sheng
FS
Chen
Jiong-Ran
JR
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D053938:DNA Fragmentation; D005434:Flow Cytometry; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D018655:Lymphocyte Count; D008214:Lymphocytes / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D011134:Polysaccharides / Q000494:pharmacology*; D031987:Potentilla; D013154:Spleen / Q000166:cytology
D011134:Polysaccharides; D006861:Hydrogen Peroxide