| Object:Lily-Rehmannia decoction has been used by Chinese to heal Lily disease for more than1,800years. In this study, we compared ultrastructure, total protein, ATP, cell membrane potential in PC-12cells treated by serum deprivation and Lily-Rehmannia decoction, to investigate the neuroprotective effects of Lily-Rehmannia decoction.Methods:(1) Lily-Rehmannia decoction:The proportion of Lilium:Rehmannia is4:3.(2) Cell line:PC-12(3) Treatment:Serum deprivation(sd),or Lily-Rehmannia decoction(LRd).(4) Groups:Cells were classified four groups:normal (control), normal+LRd, sd, sd+LRd.(5) MTT was applied to estimate suitable time and concentration of LRd.(6) Microscope for cell structure.(7) Transmission electron microscope for ultrastructure.(8) BCA method to measure total protein.(9) Fireflies luciferase method for ATP measurement.(10) Cell membrane potential was marked by detector diBAC4(3), then use Laser Scanning Confocal Microscope to compare fluorescence intensity.(11) Statistics.Results:(1)The suitable concentration of LRd was3mg/ml, treatment time was24hours.(2)Microscope observation:The majority of sd group cell became smaller and constricted, sd+LRd group cells were better in general.(3)Ultrastructure comparation:Many pinocytotic vesicle in sd group cells, some have autophagic vacuole, showed nutrition deficiency in cell; rough endoplasmic reticulum ribosomes shed, mitochondrion less, nucleus constricted, chromatin aggregate,showed disorder of protein synthesis and cell injury. Sd+LRd group cells were better, showed nutrition improved by LRd treatment.(4)Total protein:sd+LRd group total protein was more than sd group, showed that LRd could prevent the degradation of protein when nutrition deficiency.(5)ATP:The ratio of ATP/protein of sd+LRd group was more than sd group, showed that LRd could be able to improve cell’s energy level.(6)Cell membrane potential:sd group cells were brighter, with higher fluorescence intensity, showed smaller membrane potential. Sd+LRd group cells have weak fluorescence, showed bigger membrane potential.Conclusion:(1) Lily-Rehmannia decoction can improve the ultrastructure, total protein, ATP and cell membrane potential of PC-12cells treated with serum deprivation, suggest its considerable neurotrophic and neuroprotective effect.(2) Lily-Rehmannia decoction can help serum deprived PC-12cells maintaining bigger cell membrane potential, suggest that this formula could lower cell’s excitability.(3) For PC-12cells cultured with serum, Lily-Rehmannia decoction does not have distinguished cell proliferation effect. |
PDF Full Text Request