[an error occurred while processing this directive]

Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (11): 1061-1066.doi: 10.3736/jcim20091108

• Original Experimental Research • Previous Articles     Next Articles

Effects of cembrane-type diterpenes on proliferation of PC12 cells and their antagonistic effects on neurotoxicity induced by glutamate

Dong-xiao Wanga,Ping Liua,Hao-yang Rena,Wen-han Linb,Ya-qing Yanga,Xiao-fei Maa,Ting Wenb, Hong-bo Liaoa   

  1. a Drug Supply Center, General Hospital of Peopled Liberation Army of China, Beijing 100853, China
    b State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Ceter, Beijing 100083, China
  • Received:2009-06-22 Accepted:2009-09-18 Online:2009-11-20 Published:2009-11-15
  • Contact: Ping Liu E-mail:liuping126@126.com

RichHTML

2

PDF (PC)

68

Like

Objective

To investigate the effects of cembrane-type diterpenes extracted from Sinularia flexibilis on the proliferation of PC12 cells and their protective effects on PC12 cells exposed to glutamate.
Methods

Methyl thiazolyl tetrazolium (MTT) method was adopted to observe the effects of cembrane-type diterpenes (compound 1, compound 2 and compound 3) on the proliferation of PC12 cells. And the protective effects of the three compounds on PC12 cells exposed to glutamate were also detected by MTT. Furthermore, the influence of compound 1 on intracellular concentration of calcium in PC12 cells exposed to glutamate was detected by laser confocal microscopy. Results: After 72-hour PC12 cell culture, OD values in the 2, 10 and 50 μmol/L compound 1 groups were significantly higher than that in the normal group (P < 0.05, P < 0.01). After 24-hour glutamate damage, OD values in the 0.4, 2 and 50 μmol/L compound 1 groups, the 0.4, 2 and 100 μmol/L compound 2 groups and the 2 μmol/L compound 3 group were obviously increased as compared with the untreated group (P < 0.01, P < 0.05). After 48-hour glutamate damage, OD values in the compound 1 group were approximate to those in the normal control and the positive control group while were significantly higher than that in the untreated group (P < 0.01, P < 0.05), but no dose-dependent effect was observed. Compound 1 of 0.4, 2, 50 μmol/L could significantly reduce the intracellular concentration of calcium in PC12 cells exposed to glutamate (P < 0.05, P < 0.01), which was also approximate to the effect of nimodipine (positive control drug).
Conclusion

Cembrane-type diterpenes (compound 1, compound 2 and compound 3) extracted from Sinularia flexibilis have obvious protective effects on PC12 cells damaged by glutamate, and compound 1 has the best neuroprotective effect. The mechanism of the neuroprotective effect of compound 1 may lie in reducing the intracellular concentration of calcium in PC12 cells exposed to glutamate and relieving the calcium overload.

Key words: Sinularia flexibilis, Cembrane-type diterpenes, PC12 cell, Glutamate, Calcium

Figure 1

Structure formula of the compounds isolated form Sinularia flexibilis"

Table 1

Effects of cembrane-type diterpenes extracted from Sinularia flexibilis on proliferation of PC12 cells ($\bar{x}$±s)"

Group Dose (μmol/L) n OD value
24 h 48 h 72 h
Normal control 0.4 6 0.44±0.08 0.60±0.04 0.70±0.07
Compound 1 2 6 0.75±0.09** 0.70±0.05 0.85±0.16
10 6 0.54±0.01 0.80±0.28** 0.93±0.20*
50 6 0.44±0.10 0.70±0.04 0.97±0.21*
100 6 0.78±0.39** 0.89±0.28** 1.07±0.23**
0.4 6 0.55±0.01 0.66±0.10 0.83±0.09
Compound 2 2 6 0.42±0.06 0.70±0.02 0.85±0.12
10 6 0.49±0.07 0.70±0.01 0.92±0.18
50 6 0.45±0.12 0.66±0.05 0.86±0.08
100 6 0.26±0.10 0.67±0.11 0.83±0.19
0.4 6 0.45±0.08 0.45±0.12 0.45±0.10
Compound 3 2 6 0.55±0.14 0.67±0.10 0.88±0.21
10 6 0.55±0.01 0.74±0.05 0.77±0.05
50 6 0.50±0.08 0.73±0.02 0.92±0.22
100 6 0.50±0.07 0.45±0.10 0.42±0.10
6 0.52±0.02 0.55±0.05 0.48±0.13

Table 2

Effects of cembrane-type diterpenes extracted from Sinularia flexibilis on PC12 cells exposed to glutamate ($\bar{x}$±s)"

Group Dose (μmol/L) n OD value
24 h 48 h
Normal control 6 0.54±0.07** 0.85±0.07**
Untreated 500 6 0.34±0.02 0.71±0.02
Positive control 5 6 0.55±0.09** 0.83±0.09**
Compound 1 0.4 6 0.46±0.10** 0.83±0.09*
2 6 0.49±0.05** 0.84±0.10**
10 6 0.37±0.02 1.01±0.14**
50 6 0.49±0.07* 0.82±0.05*
100 6 0.36±0.04 0.84±0.13**
Compound 2 0.4 6 0.45±0.04** 0.75±0.07
2 6 0.49±0.06** 0.76±0.05
10 6 0.40±0.03 0.77±0.09
50 6 0.39±0.04 0.75±0.06
100 6 0.49±0.06** 0.70±0.06
Compound 3 0.4 6 0.35±0.10 0.74±0.05
2 6 0.42±0.03* 0.71±0.06
10 6 0.34±0.10 0.71±0.05
50 6 0.40±0.03 0.72±0.06
100 6 0.38±0.03 0.68±0.11

Figure 2

Effects of compound 1 on intracellular calcium concentration in PC12 cell exposed to glutamate A: Normal control group; B: Untreated group; C: Positive control group; D-H: Different concentrations of compound 1(0.4, 2, 10, 50 and 100 μmol/L) respectively. Data were represented as $\bar{x}$±s, n=6. *P<0.05, **P<0.01, vs untreated group."

[1] Li GQ, Zhang YL, Lin WH . Research progress on the cembranoid diterpenes from oceanic organisms[J]. Zhongguo Hai Yang Da Xue Xue Bao, 2006,36(3):370-376
李国强, 张艳玲, 林文翰 . 西松烷二萜类海洋活性成分研究进展[J]. 中国海洋大学学报, 2006,36(3):370-376
[2] Wen T, Liu WW, Lin WH . Study on cembrane-type diterpenes in soft coral Sinularia flexibilis collected from South China Sea[J]. Zhongguo Yao Xue Za Zhi, 2008,43(7):493-496
温烃, 刘文威, 林文翰 . 中国南海软珊瑚Sinularia flexibilis中西松烷型二萜的研究[J]. 中国药学杂志, 2008,43(7):493-496
[3] Yan XH, Guo YW . A pharmaceutical perspective on soft coral: chemistry and bioactivity[J]. Zhongguo Tian Ran Yao Wu, 2005,3(2):65-73
严小红, 郭跃伟 . 软珊瑚化学成分和生物活性的研究进展[J]. 中国天然药物, 2005,3(2):65-73
[4] Song MX, Yang JC, Wang WP . A study of PC12 cell damage induced by glutamate[J]. Suzhou Ke Ji Xue Yuan Xue Bao, 2006,23(1):62-65
doi: 10.3969/j.issn.1672-0687.2006.01.016
宋明旭, 杨吉成, 王蔚平 . 谷氨酸诱导的PC12细胞损伤的研究[J]. 苏州科技学院学报, 2006,23(1):62-65
doi: 10.3969/j.issn.1672-0687.2006.01.016
[5] Wu Y, Zhang YJ, Miao QF, Xue JM, Gao XF . Protective effects of dipfluzine on glutamate induced neurotoxicity in cultured hippocampal neurons[J]. Ningxia Yi Ke Da Xue Xue Bao, 2009,31(1):1-3
吴洋, 张永健, 苗庆峰, 薛健梅, 高霄飞 . 双苯氟嗪对谷氨酸致海马神经元损伤的保护作用[J]. 宁夏医科大学学报, 2009,31(1):1-3
[6] Huang JS, Long LJ, Zhang S . Progress in the research on marine natural products and their physiological activities[J]. Hai Yang Tong Bao, 2001,20(4):83-91
黄建设, 龙丽娟, 张偲 . 海洋天然产物及其生理活性的研究进展[J]. 海洋通报, 2001,20(4):83-91
[7] Ouyang GL, Li QF, Tian CH . Progress in studies on marine antitumor bioactive substances and their antitumor mechanisms[J]. Hai Yang Ke Xue, 2003,27(7):21-24
doi: 10.3969/j.issn.1000-3096.2003.07.006
欧阳高亮, 李祺福, 田长海 . 海洋生物抗肿瘤活性物质及其作用机理研究进展[J]. 海洋科学, 2003,27(7):21-24
doi: 10.3969/j.issn.1000-3096.2003.07.006
[8] Peruche B, Krieglstein J . Neuroblastoma cells for testing neuroprotective drug effects[J]. J Pharmacol Methods, 1991,26(2):139-148
doi: 10.1016/0160-5402(91)90062-A pmid: 1943124
[9] Shafer TJ, Atchison WD . Transmitter, ion channel and receptor properties of pheochromocytoma(PC12) cells: a model for neurotoxicological studies[J]. Neurotoxicology, 1991,12(3):473-492
[10] Michaelis EK . Molecular biology of glutamate receptors in the central nervous system and their role in excitotoxicity, oxidative stress and aging[J]. Prog Neurobiol, 1998,54(4):369-415
doi: 10.1016/S0301-0082(97)00055-5
[11] Paschen W . Role of calcium in neuronal cell injury: which subcellular compartment is involved?[J]. Brain Res Bull, 2000,53(4):409-413
doi: 10.1016/S0361-9230(00)00369-5
[12] Rameau GA, Akaneya Y, Chiu L, Ziff EB . Role of NMDA receptor functional domains in excitatory cell death[J]. Neuropharmacology, 2000,39(12):2255-2266
doi: 10.1016/S0028-3908(00)00066-6 pmid: 10974309
[13] Biagas K . Hypoxic-ischemic brain injury: advancements in the understanding of mechanisms and potential avenues for therapy[J]. Curr Opin Pediatr, 1999,11(3):223-228
doi: 10.1097/00008480-199906000-00009
[1] Xiao-ping Huang , Xiao-dan Liu , Chang-qing Deng . Effects of the combination of active component extracts from Astragalus membranaceus and Panax notoginseng on apoptosis, reactive oxygen species and mitochondrial membrane potential of PC12 cells with oxidative injury. Journal of Chinese Integrative Medicine, 2012, 10(10): 1127-1134.
[2] Cheng Jian, Zhang Xin-min, Shen Zi-yin, Chen Wei-hua, Cai Wai-jiao, Ying Jian, Hu Guo-rang, Liu Run-hong. Immunoregulatory mechanisms of an optimal Chinese herbal monomer compound in mice with allergic rhinitis. Journal of Chinese Integrative Medicine, 2011, 9(12): 1360-1366.
[3] Wen-jun Wang, Da-jin Li, Jun Li, Wen-jiang Zhou. An in vitro study on neuroprotective effects of serum containing Gengnianchun decoction and its main monomers against amyloid beta protein-induced cellular toxicity. Journal of Chinese Integrative Medicine, 2010, 8(1): 67-73.
[4] Pei Wang, Li-ping Xu, Xi-hong Jin, Jing Yang, Ren-hui Liu, Xiu-juan Wang . Effects of a glucocorticoid on development of kidney deficiency syndrome in a rat model of asthma. Journal of Chinese Integrative Medicine, 2010, 8(1): 80-85.
[5] Ning Chen , Hui Li , Li-yun Zhao , Jian-bo Liu , Qin Huang , Tong-han Wang , Xiao-mei Hao . Therapy for clearing heat and resolving phlegm in treatment of systemic inflammatory response syndrome in acute deterioration stage of chronic obstructive pulmonary disease: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2009, 7(2): 105-109.
[6] Jian-ye Yuan, Jian-qun Xie, Da-zheng Wu, Yu Zheng, Xiang-xue Pan, Xiao-yan Fei, Hai-zhen Xu. Tongxie Yaofang inhibits the contraction of colonic smooth muscle isolated from rats through a mechanism related to calcium mobilization. Journal of Chinese Integrative Medicine, 2009, 7(10): 958-962.
[7] Li-hui Dang, Xiao-nan Yang, Qing Xia. Protective effects of Chaiqin Chengqi Decoction on isolated pancreatic acinar cells in acute pancreatitis rats and the mechanisms. Journal of Chinese Integrative Medicine, 2008, 6(2): 176-179.
[8] Ping Xue, Li-hui Deng, Zhao-da Zhang, Xiao-nan yang, Qing Xia, Da-kai Xiang, Lei Huang, Mei-hua Wan, Hai-yan Zhang. Chaiqin Chengqi Decoction decreases pancreatic acinar cell calcium overload in rats with acute pancreatitis. Journal of Chinese Integrative Medicine, 2008, 6(10): 1054-1058.
[9] Rui-wu Li, Jin-ling Zhang, Ying Guo, Cui-hong Li. Effect of acupuncture serum on intracellular Ca2+ concentration in cultured neurons. Journal of Chinese Integrative Medicine, 2004, 2(6): 453-455.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[2] Hao Li, Ming-jiang Yao, Wen-ming Zhao, Jie Guan, Lin-lin Cai, Ling Cui. A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment. Journal of Chinese Integrative Medicine, 2008, 6(1): 25-31
[3] Zhi-chun Jin. Problems in establishing clinical guideline for integrated traditional Chinese and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 5-8
[4] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[7] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[8] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[9] Gui Yu, Jie Wang. Thinking on building the network cardiovasology of Chinese medicine. Journal of Chinese Integrative Medicine, 2012, 10(11): 1206-1210
[10] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239
[an error occurred while processing this directive]