[Effects of cembrane-type diterpenes on proliferation of PC12 cells and their antagonistic effects on neurotoxicity induced by glutamate].
OBJECTIVE
To investigate the effects of cembrane-type diterpenes extracted from Sinularia flexibilis on the proliferation of PC12 cells and their protective effects on PC12 cells exposed to glutamate.


METHODS
Methyl thiazolyl tetrazolium (MTT) method was adopted to observe the effects of cembrane-type diterpenes (compound 1, compound 2 and compound 3) on the proliferation of PC12 cells. And the protective effects of the three compounds on PC12 cells exposed to glutamate were also detected by MTT. Furthermore, the influence of compound 1 on intracellular concentration of calcium in PC12 cells exposed to glutamate was detected by laser confocal microscopy.


RESULTS
After 72-hour PC12 cell culture, OD values in the 2, 10 and 50 micromol/L compound 1 groups were significantly higher than that in the normal group (P<0.05, P<0.01). After 24-hour glutamate damage, OD values in the 0.4, 2 and 50 micromol/L compound 1 groups, the 0.4, 2 and 100 micromol/L compound 2 groups and the 2 micromol/L compound 3 group were obviously increased as compared with the untreated group (P<0.01, P<0.05). After 48-hour glutamate damage, OD values in the compound 1 group were approximate to those in the normal control and the positive control group while were significantly higher than that in the untreated group (P<0.01, P<0.05), but no dose-dependent effect was observed. Compound 1 of 0.4, 2, 50 micromol/L could significantly reduce the intracellular concentration of calcium in PC12 cells exposed to glutamate (P<0.05, P<0.01), which was also approximate to the effect of nimodipine (positive control drug).


CONCLUSIONS
Cembrane-type diterpenes (compound 1, compound 2 and compound 3) extracted from Sinularia flexibilis have obvious protective effects on PC12 cells damaged by glutamate, and compound 1 has the best neuroprotective effect. The mechanism of the neuroprotective effect of compound 1 may lie in reducing the intracellular concentration of calcium in PC12 cells exposed to glutamate and relieving the calcium overload.
Wang
Dong-xiao
DX
Drug Supply Center, General Hospital of Liberation Army of China, Beijing 100853, China.
Liu
Ping
P
Ren
Hao-yang
HY
Lin
Wen-han
WH
Yang
Ya-qing
YQ
Ma
Xiao-fei
XF
Wen
Ting
T
Liao
Hong-bo
HB
D000818:Animals; D037421:Anthozoa / Q000737:chemistry; D049109:Cell Proliferation / Q000187:drug effects*; D004224:Diterpenes / Q000302:isolation & purification / Q000494:pharmacology*; D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates / Q000633:toxicity; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016716:PC12 Cells; D051381:Rats
D004224:Diterpenes; D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates; D018696:Neuroprotective Agents