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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (5): 472-479.doi: 10.3736/jcim20100512

• Original Experimental Research • Previous Articles     Next Articles

Medicated rat serum containing Gengnianchun decoction reduces apoptosis of pheochromocytoma cells insulted by amyloid beta protein

 Jun Lia, Bin Lia, Wen-jiang Zhoub, Fan-gui Zhaoa, Da-jin Lia, Wen-jun Wanga   

  1. a Department of Integrated Traditional Chinese and Western Medicine,Gynecology and Obstetrics Hospital, Fudan University, Shanghai 200011, China
    b Department of Experimental Animal, Fudan University, Shanghai 200023, China
  • Received:2009-12-31 Accepted:2010-03-08 Online:2010-05-20 Published:2010-05-15
  • Contact: Wen-jun Wang


To investigate the effects of medicated rat serum containing Gengnianchun (GNC) decoction and its protection to pheochromocytoma cells (PC12 cells) from amyloid beta (Aβ)25-35-insulted apoptosis and to find the possible mechanism.

Medicated rat serum was prepared by administering ovariectomized Sprague-Dawley (SD) rats with GNC decoction. The effects of medicated rat serum on viability of PC12 cells were evaluated by cell counting kit-8 (CCK-8) assay. The PC12 cells were cultured with different doses of Aβ25-35 to induce a model of Alzheimer's disease in vitro. Then, the protective effects of medicated rat serum on Aβ25-35-insulted PC12 cells were evaluated by using CCK-8 assay to detect the cell viability, using Annexin Ⅴ-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry to detect cell apoptosis rate and using Western blotting assay to analyze the expressions of Bcl-2, Bax and active caspase-3 proteins.

PC12 cells cultured with 20% medicated rat serum containing GNC decoction for 24 h or 48 h had higher viability than those cultured with normal culture medium (P<0.05). After 24- or 48-hour treatment of different concentrations of Aβ25-35, cell viabilities were all decreased as compared with normal medium (P<0.05). Cells underwent apoptosis, which showed the neurotoxicity of Aβ25-35. The cell apoptosis induced by Aβ25-35 was significantly decreased in PC12 cells which were pretreated with 20% medicated rat serum or nerve growth factor (NGF) according to CCK-8 assay and Annexin Ⅴ-FITC/PI flow cytometry (P<0.05). The ratio of Bax expression to Bcl-2 expression and the expression of active caspase-3 were decreased in the cells treated with medicated serum or NGF as compared with the cells cultured with Aβ25-35 only.

The GNC-medicated rat serum at concentration of 20% can promote viability of Aβ25-35-insulted PC12 cells and decrease the cell apoptosis by regulating the expressions of Bcl-2, Bax and active caspase 3.

Key words: Alzheimer’s disease, Apoptosis, Medicated serum, Amyloid beta, Bcl-2

Figure 1

Proliferation rate of PC12 cells cultured with medicated serum containing GNC or non-medicated rat serum"

Figure 2

Cytotoxicity of Aβ25-35 to PC12 cells"

Figure 3

Protective effects of medicated rat serum on Aβ25-35-induced cytotoxicity in PC12 cells"

Figure 4

Protective effects of medicated rat serum on Aβ25-35-induced apoptosis in PC12 cells"

Figure 5

Preventive effects of medicated serum on Aβ25-35-induced apoptotic signaling"

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