Open AccessEffects of huperzine A on secretion of nerve growth factor in cultured rat cortical astrocytes and neurite outgrowth in rat PC12 cells 1
Author(s) -
TANG Lili,
WANG Rui,
TANG Xican
Publication year - 2005
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2005.00130.x
Subject(s) - neurite , nerve growth factor , huperzine a , cytotoxicity , lactate dehydrogenase , messenger rna , microbiology and biotechnology , viability assay , acetylcholinesterase , western blot , chemistry , medicine , endocrinology , biology , receptor , cell , enzyme , biochemistry , in vitro , gene
Abstract Aim: 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 μmol/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's 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 μmol/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 μmol/ L HupA also increased the mRNA and protein levels of this enzyme. In addition, following 2 h exposure of the astrocytes to 10 μmol/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. Conclusion: 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.