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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 E-mail:abby_lijun@163.com

Objective

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.
Methods

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.
Results

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.
Conclusion

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"

[1] Zheng WH, Bastianetto S, Mennicken F, Ma W, Kar S . Amyloid beta peptide induces tau phosphorylation and loss of cholinergic neurons in rat primary septal cultures[J]. Neuroscience, 2002,115(1):201-211
doi: 10.1016/S0306-4522(02)00404-9
[2] Burrerfield DA, Castegna A, Lauderback CM, Drake J . Evidence that amyloid β-peptide-induced lipid peroxidation and its sequelae in Alzheimer's disease brain contribute to neuronal death[J]. Neurobiol Aging, 2002,23(5):655-664
doi: 10.1016/S0197-4580(01)00340-2
[3] Nakagawa T, Zhu H, Morishima N, Li E, Xu J, Yankner BA, Yuan J . Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta[J]. Nature, 2000,403(6765):98-103
doi: 10.1038/47513
[4] Cao LX, Mao QZ, Yu J . Clinical research of Gengnianchun decoction in treatment of perimenopausal syndrome[J]. Shanghai Zhong Yi Yao Za Zhi, 1997,31(2):33-35
曹玲仙, 毛秋芝, 俞瑾 . 更年春治疗更年期综合征的临床研究[J]. 上海中医药杂志, 1997,31(2):33-35
[5] Jing SY, Wei MJ . Clinical observation of “Climacterium capsule” in improving the memory of climacteric women[J]. Shanghai Zhong Yi Yao Za Zhi, 2001,35(8):32-33
景苏玉, 魏美娟 . 更年青胶囊对提高更年期妇女记忆力的观察[J]. 上海中医药杂志, 2001,35(8):32-33
[6] Zhao FG, Wang WJ, Zhou WJ, Li DJ . Effect of Gengnianchun Recipe on learning memory function and hippocampal cholinergic system in ovariectomized rats[J]. Zhongguo Zhong Xi Yi Jie He Za Zhi, 2008,28(3):234-237
赵凡桂, 王文君, 周文江, 李大金 . 更年春方对切除卵巢大鼠学习记忆能力和海马胆碱能系统的影响[J]. 中国中西医结合杂志, 2008,28(3):234-237
[7] Wang XP, Ding HL . Alzheimer's disease: epidemiology, genetics, and beyond[J]. Neurosci Bull, 2008,24(2):105-109
doi: 10.1007/s12264-008-0105-7
[8] Lopez OL, Jagust WJ , DeKosky ST, Becker JT, Fitzpatrick A, Dulberg C, Breitner J, Lyketsos C, Jones B, Kawas C, Carlson M, Kuller LH .Prevalence and classification of mild cognitive impairment in the Cardiovascular Health Study Cognition Study: part 1[J]. Arch Neurol, 2003,60(10):1385-1389
doi: 10.1001/archneur.60.10.1385
[9] Zandi PP, Carlson MC, Plassman BL, Welsh-Bohmer KA, Mayer LS, Steffens DC, Breither JC , Cache County Memory Study Investigators.Hormone replacement therapy and incidence of Alzheimer disease in older women: the Cache Country Study[J]. JAMA, 2002,288(17):2123-2129
doi: 10.1001/jama.288.17.2123
[10] Li M, Chen L, Lee DH, Yu LC, Zhang Y . The role of intracellular amyloid beta in Alzheimer's disease[J]. Prog Neurobiol, 2007,83(3):131-139
doi: 10.1016/j.pneurobio.2007.08.002
[11] Gross A , McDonnell JM, Korsmeyer SJ.Bcl-2 family members and the mitochondria in apoptosis[J]. Genes Dev, 1999,13(15):1899-1911
doi: 10.1101/gad.13.15.1899
[12] Suh YH, Checler F . Amyloid precursor protein, presenilins, and alpha-synuclein: molecular pathogenesis and pharmacological applications in Alzheimer's disease[J]. Pharmacol Rev, 2002,54(3):469-525
doi: 10.1124/pr.54.3.469
[13] Kang SY, Lee KY, Koo KA, Yoon JS, Lim SW, Kim YC, Sung SH . ESP-102, a standardized combined extract of Angelica gigas, Saururus chinensis and Schizandra chinensis, significantly improved scopolamine-induced memory impairment in mice[J]. Life Sci, 2005,76(15):1691-1705
doi: 10.1016/j.lfs.2004.07.029
[14] Yang F, Lim GP, Begum AN, Ubeda OJ, Simmons MR, Ambegaokar SS, Chen PP, Kayed R, Glade CG, Frautschy SA, Cole GM . Curcumin inhibits formation of amyloid beta oligomers and fibrils, binds plaques, and reduces amyloid in vivo[J]. J Biol Chem, 2005,280(7):5892-5901
doi: 10.1074/jbc.M404751200
[15] Zhao FG, Wang WJ, Zhou WJ, Li DJ . Regulative effects of “Gengnianchun decoction” on learning and memory and hippocampal ER subtype in ovariectomized rats[J]. Shanghai Zhong Yi Yao Za Zhi, 2007,41(11):66-69
赵凡桂, 王文君, 周文江, 李大金 . 更年春方对卵巢切除大鼠学习记忆能力和海马ER亚型的调节[J]. 上海中医药杂志, 2007,41(11):66-69
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