Effects of huperzine A on secretion of nerve growth factor in cultured rat cortical astrocytes and neurite outgrowth in rat PC12 cells.
OBJECTIVE
To study the effects of huperzine A (HupA) on neuritogenic activity and the expression of nerve growth factor (NGF).


METHODS
After being treated with 10 micromol/L HupA, neurite outgrowth of PC12 cells was observed and counted under phase-contrast microscopy. Mitogenic activity was assayed by [3H]thymidine incorporation. Cell cytotoxicity was evaluated by lactate dehydrogenase (LDH) release. AChE activity, mRNA and protein expression were measured by the Ellman method, RT-PCR, and Western blot, respectively. NGF mRNA and protein levels were determined by RT-PCR and ELISA assays.


RESULTS
Treatment of PC12 cells with 10 micromol/L HupA for 48 h markedly increased the number of neurite-bearing cells, but caused no significant alteration in cell viability or other signs of cytotoxicity. In addition to inhibiting AChE activity, 10 micromol/L HupA also increased the mRNA and protein levels of this enzyme. In addition, following 2 h exposure of the astrocytes to 10 micromol/L HupA, there was a significant up-regulation of mRNA for NGF and P75 low-affinity NGF receptor. The protein level of NGF was also increased after 24 h treatment with HupA.


CONCLUSIONS
Our findings demonstrate for the first time that HupA has a direct or indirect neurotrophic activity, which might be beneficial in treatment of neurodegenerative disorders such as Alzheimer disease.
Tang
Li-li
LL
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China.
Wang
Rui
R
Tang
Xi-can
XC
D000110:Acetylcholinesterase / Q000096:biosynthesis* / Q000235:genetics; D000470:Alkaloids; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D032501:Huperzia / Q000737:chemistry; D020932:Nerve Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D016501:Neurites / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology; D016716:PC12 Cells; D010946:Plants, Medicinal / Q000737:chemistry; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012717:Sesquiterpenes / Q000302:isolation & purification / Q000494:pharmacology*
D000470:Alkaloids; D002800:Cholinesterase Inhibitors; D018696:Neuroprotective Agents; D012333:RNA, Messenger; D012717:Sesquiterpenes; C050426:huperzine A; D020932:Nerve Growth Factor; D000110:Acetylcholinesterase