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Journal of Chinese Integrative Medicine: 2010; 8(1): 67-73
DOI: 10.3736/jcim20100113
An in vitro study on neuroprotective effects of serum containing Gengnianchun decoction and its main monomers against amyloid beta protein-induced cellular toxicity
1. Wen-jun WANG (Department of Integrated Traditional Chinese and Western Medicine, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China )
2. Da-jin LI (Department of Integrated Traditional Chinese and Western Medicine, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China )
3. Jun LI (Department of Integrated Traditional Chinese and Western Medicine, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China )
4. Wen-jiang ZHOU (Department of Experimental Animal, Fudan University, Shanghai 200023, China )

Obiective: To observe the effects of serum containing Gengnianchun (GNC) decoction, a compound traditional Chinese herbal medicine, and its monomers (paeoniflorin, berberine, timosaponin A-Ⅲ and icariin) on neurotoxicity in PC12 cells induced by amyloid beta-protein (Aβ).
Methods: Injury of PC12 cells was induced by in incubating with Aβ25-35 in vitro. Ovariectomized rats were intragastrically administered with GNC decoction twice daily for 5 days and then sera were obtained. Different concentrations of serum containing GNC decoction and its main monomers including paeoniflorin, berberine, timosaponin and icariine and the monomer mixtures were cultured with PC12 cells to determine the best concentration of the drugs by methyl thiazolyl tetrazolium (MTT) method. The effective concentration of Aβ25-35 was detemined by culturing PC12 cells with different concentrations of Aβ25-35. Then, the activity of PC12 cells with Aβ25-35-induced injury was observed with MTT method. Cellular morphological change was observed by phase contrast microscopy. Flow cytometry and fluorescence microscopy were employed to observe the Aβ25-35-induced early apoptosis of PC12 cells.
Results: After Aβ25-35 induction, the PC12 cells were fewer in number, less viable with shrinked cel1 body, many fragments, adhered less and nuclei shrinked. The cell proliferation was inhibited by Aβ25-35 concentration- and timedependently. Aβ25-35 at concentration of 20 μmol/ L was selected to construct the Alzheimer's disease model in vitro. The sera containing GNC decoction could reinforce PC12 cell activity, and concentration at 20% was better than other concentrations after 24-, 48- and 72-hour culture. The 20% serum containing GNC decoction, 0.1 μmol/L  berberin and monomer mixture 3 including 1 mg/mL paeoniflorin, 1 μmol/L berberine, 1 μmol/L timosaponin and 1 ng/mL icariine could antagonize neurotoxicity induced by Aβ25-35. Moreover, they could inhibit Aβ25-35-induced early apoptosis of PC12 cells, with the effect of 20% serum containing GNC decoction better than 0.1 μmol/L berberine and monomer mixture 3.
Conclusion: Serum containing GNC decoction at 20% concentration has the potential neuroprotective effect on Aβ-induced cellular impairment. The serum containing GNC decoction was found to be stronger in action than the main monomers.

 

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Please cite this article as:
Wang WJ, Li DJ, Li J, Zhou WJ. An in vitro study on neuroprotective effects of serum containing Gengnianchun decoction and its main monomers against amyloid beta proteininduced cellular toxicity. J Chin Integr Med / Zhong Xi Yi Jie He Xue Bao. 2010; 8(1): 67-73.
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