OBJECTIVE To investigate the
puerarin induced VSMC
apoptosis gene and the molecular mechanism of this effect on anti-atherosclerosis.METHODS The RNA of both the control VSMCs and those stimulated by puerarin were isolated and analyzed by differential display RT-PCR (DDRT-PCR).These differentially expressed fragments were recovered,cloned,sequenced and compared with GeneBank databases.One
est (expressional Sequence Tag) was selected to further analyzed by northern
hybridization and
tissue situ hybridization to observe the
gene expression of the GRP94 in VSMC of apoptosis and AS.RESULTS One EST,733 bp,was isolated from VSMC stimulated by puerarin,similar to the gene of glucose regulated protein GRP94.The
expression of the EST in VSMC was correlated to the concentration of puerarin and the level of expression was significantly higher in high cholesterol group than in control group.CONCLUSION The GRP94 gene expressed in both stressed model of puerarin-induced apoptosis and high cholesterol induced AS.Puerarin possesses the function of promoting apoptosis of VSMC.Its molecular mechanism may be related to promoting the gene expression of the GRP94 of VSMC.
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