Inhibitory Actions of Tetrandrine on Tumor Necrosis Factor α-Induced NF-κB Activation in Neovascularization of Cultured Choroidal Explants


Tetrandrine (1 μM), a bis-benzylisoquinoline alkaloid isolated from Stephania tetrandra S Moore, signifi-cantly decreased tumor necrosis factor alpha (TNFα; 10 ng/ml)-induced increase in the number of micro vessels that budded from cultured rat choroidal explants. Tetrandrine also decreased the TNFα-induced in-crease in the number of cells composing the microvessels. Ammonium pyrrolidine dithiocarbamate (APDC; 0.1-0.3 μM), an inhibitor of nuclear factor-κB (NF-κB), decreased the TNFα-induced increase in the number of microvessels in a concentration-dependent manner. TNFα increased the phosphorylation and degradation of inhibitor of NF-κB (IκBα), as well as increasing the DNA-binding activity of NF-κB in choroidal explants. TNF? induced an increase of vascular endothelial growth factor (VEGF)-A mRNA, but not VEGF-C mRNA or VEGF-D mRNA. TNFα-induced angiogenic action was inhibited by treatment of VEGF-A antibody in cultured choroidal capillaries. Tetrandrine inhibited the TNFα-induced increases of phosphorylation and degradation of IκBα, and reduced the TNFα-induced increase of DNA-binding activity of NF-κB in chor-oidal explants. In conclusion, tetrandrine inhibits TNFα-induced activation of NF-κB in the choroidal capil-laries via inhibition of TNFα-induced phosphorylation of IκBα.

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M. Kikuchi, S. Kamimura, M. Nomura, T. Takahashi, N. Hagino and S. Kobayashi, "Inhibitory Actions of Tetrandrine on Tumor Necrosis Factor α-Induced NF-κB Activation in Neovascularization of Cultured Choroidal Explants," Chinese Medicine, Vol. 1 No. 3, 2010, pp. 75-83. doi: 10.4236/cm.2010.13015.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] X. X. Yu, J. X. Dang, L. Shi, P. F. Gu, X. H. Xu and F. Z. Kong, “Clinical Observation of Tetrandrine Aero Sol on the Effect of Relieving Asthma,” Zhejiang Journal Traditional Chinese Medicine, Vol. 27, 1992, pp. 98-99.
[2] D. Li, H. Lu, X. Li, Q. Ouan, X. Li and W. Lu, “Calcium Channel Blockers in Cirrhotic Patients with Portal Hypertension,” Chinese Medicine Journal (Engl), Vol. 108, 1995, pp. 803-808.
[3] D. H. Li, C. J. Zhang and Y. N. Zhong, “Clinical Study on Long-Term Therapeutic Effect of Combined Anti-Fibrosis, 2001,” Chin J Indus Hygiene Occup Dis , Vol. 19, 2001, pp. 3-6.
[4] Q. M. Xie, H. F. Tang, J. Q. Chen and R. L. Bian, “Pharmacological Actions of Tetrandrine in Inflammatory Pulmonary Disease,” Acta Pharmacologica Sinica, Vol. 23, 2002, pp. 1107-1113.
[5] C. W. Wong, W. K. Seow, J. W. O’Callaghan and Y. H. Thong, “Comparative effects of Tetrandrine and Berbamine on Subcutaneous Air Pouch Inflammation Induced by Interleukin-1, Tumor Necrosis Factor and Platelet-Activating Factor,” Agents Actions, Vol. 36, 1992, pp. 112- 118.
[6] C. W. Wong, Y. H. Thong and W. K. Seow, “Comparative Effects of Tetrandrine and Berbamine on Guinea Pig Airway Micro Vascular Leakage Induced by Platelet-Activating Factor and Other Allergic Mediators,” International Journal of Immunopharmacology, Vol. 15, 1993, pp. 185-193.
[7] K. Takahashi, H. Kobayashi, S. Kobayashi, I. Kimura, K. Terasawa and M. Kimura, “Antiproliferative Effects of Magnosalin Derived from ‘Shin’i’ (Flos Magoliae), a Japanese Sino-Medicine, on Cultured Synovial Cells of MRL/l and C57BL/6J Mice,” Phototherapy Research, Vol. 10, 1996, pp. 42-48.
[8] S. Kobayashi, K. Inaba, I. Kimura and M. Kimura, “Inhibitory Effects of Tetrandrine on Angiogenesis in Adjuvant-Induced Chronic Inflammation and Tube Formation of Vascular Endothelial Cells,” Biological and Pharmaceutical Bulletin, Vol. 21, 1998, pp. 346-349.
[9] B. S. Teh, W. K. Seow, S. Y. Li and Y. H. Thong, “Inhibition of Prostaglandin and Leukotriene Generation by the Plant Alkaloids Tetrandrine and Berbamine,” International Journal of Immunopharmacology Vol. 12, 1990, pp. 321- 326.
[10] G. Wang and J. R. Lemos, “Tetrandrine: A New Ligand to Block Voltage-Dependent Ca2+ and Ca(2+)-Activated K+ Channels,” Life Sciences, Vol. 56, 1995, pp. 295-306.
[11] C. Y. Kwan, F. M. Ma and S. C. Hui, “Inhibition of Endothelium-Dependent Vascular Relaxation by Tetrandrine,” Life Sciences, Vol. 64, 1999, pp. 2391-2400.
[12] W. Li, W. Chen, S. Zhang and S. Lei, “Effect of Tetrandrine on Pulmonary Vascular Morphology in Rats with Hypoxia Oulmonary Hypertension,” Journal of West China University Medical Sciences, Vol. 28, 1997, pp. 389-391.
[13] G. Sun, Y. Qi and Q. Pan, “Quantitative Analysis of Prevention Effect of Tetrandrine on Pancreatic Islet Beta Cells Injury in Rats,” Chinese Medical Journal, Vol. 77, 1997, pp. 270-277.
[14] I. Lieberman, D. P. Lentz, G. A. Trucco, W. K. Seow and Y. H. Thong, “Prevention by Tetrandrine of Spontaneous Development of Diabetes Mellitus in BB Rats,” Diabetes, Vol. 41, 1992, pp. 616-619.
[15] D. O. Bates and F. E. Curry, “Vascular Endothelialgrowth Factor Increases Microvascular Permeability via A Ca(2+)-Dependent Pathway,” American Journal of Physiology Vol. 273, 1997, pp. H687-H694.
[16] K. S. Bauer, K. J. Cude, S. C. Dixon, E. A. Kruger and W. D. Figg, “Carboxyamidotriazole Inhibits Angiogenesis by Blocking the Calcium-Mediated Nitric-Oxide Synthase- Vascular Endothelial Growth Factor Pathway,” The Journal of Pharmacology and Experimental Therapeutics, Vol. 292, 2000, pp. 31-37.
[17] S. Kobayashi, I. Kimura, M. Fukuta, H. Kontani, K. Inaba, M. Niwa, S. Mita and M. Kimura, “Inhibitory Effects of Tetrandrine and Its Related Synthetic Compounds on Angiogenesis in Streptozotocin-Diabetic Rodent,” Biological and Pharmaceutical Bulletin, Vol. 22, 1999, pp. 360-365.
[18] S. Kobayashi, H. Shinohara, H. Tsuneki, R. Nagai, and S. Horiuchi, “N-(Carboxymethyl) Lysine Proliferates CD34+ Cells from Rat Choroidal Explant in Culture,” Biological and Pharmaceutical Bulletin, Vol. 27, 2004, pp. 1382-1387.
[19] S. Kobayashi, M. Suzuki, H. Tsuneki, R. Nagai, S. Horiuchi and N. Hagino, “Overproduction of N- (Carboxymethyl) Lysine-Induced Neovascularization in Cultured Choroidal Explant of Streptozotocin-Diabetic Rat,” Biological and Pharmaceutical Bulletin, Vol. 27, 2004, pp. 1565-1571.
[20] S. Kobayashi, M. Nomura, T. Nishioka, M. Kikuchi, A. Ishihara, R. Nagai and N. Hagino, “Overproduction of N?-(carboxymethyl) Lysine-Induced Neovascularization in Cultured Choroidal Explant of Aged Rat,” Biological and Pharmaceutical Bulletin, Vol. 30, 2007, pp. 133-138.
[21] S. Kobayashi, M. Nomura, T. Takahashi, M. Suzuki, R. Nagai and N. Hagino, “N?-(Carboxymethyl) Lysine- Induced Choroidal Angiogenic Potential Facilitates Retinal Neovascularization in Advanced-Diabetic Rat in Vitro,” the Open Pharmacology Journal, Vol. 2, 2008, pp. 79-86.
[22] X- C. Liang, N. Hagino, S- S., Guo, T. Tsutsumi and S. Kobayashi, “Therapeutic Efficacy of Stephania tetrandra S. Moore for Treatment of Neovascularization of Retinal Capillary (Retinopathy) in Diabetes-in Vitro Study,” Phytomedicine, Vol. 9, 2002, pp. 377-384.
[23] T. Tsutsumi, N. Hagino, X- C. Liang, S- S. Guo and S. Kobayashi, “Effects of Oral Administration of Stephania tetrandra S. Moore on Neovascularization of Retinal and Choroidal Capillaries of Diabetes in Rats,” Phototherapy Research, Vol. 22, 2008, pp 591-596.
[24] K. Hawrami, G. A. Hitman, M. Rema, C. Snehalatha, M. Viswanathan, A. Ramachandran and V. Mohan, “An Association in Non-Insulin-Dependent Diabetes Mellitus Subjects between Susceptibility,” Human Immunology, Vol. 46, 1996, pp. 49-54.
[25] G. A. Limb, A. H. Chignell, W. Green, F. LeRoy and D. C. Dumonde, “Distribution of TNF Alpha and Its Reactive Vascular Adhesion Molecules in Fibrovascular,” British Journal of Ophthalmology, Vol. 80, 1996, pp. 168-173.
[26] J. Spranger, R. Meyer-Schwickerath, M. Klein, H. Schatz and A. Pfeiffer, “TNF-Alpha Level in the Vitreous Body. Increase in Neovascular Eye Diseases and Proliferative Diabetic Retinopathy,” Medizinische Klinik (Munich, Germany), Vol. 90, 1995, pp. 134-137.
[27] M. Frater-Schroder, W. Risau, R. Hallmann, P. Gautschi and P. Bohlen, “Tumor Necrosis Factor Type Alpha, A Potent Inhibitor of Endothelial Cell Growth in Vitro, is Angiogenic in Vivo,” Proceedings of National Academy of Sciences of the United States of America, Vol. 84, 1987, pp. 5277-5281.
[28] S. J. Leibovich, P. J. Polverini, H. M. Shepard, D. M. Wiseman, V. Shively, N. Nuseir, “Macrophage-Induced Angiogenesis is Mediated by Tumor Necrosis Factor-Alpha,” Nature, Vol. 329, 1987, pp. 630-632.
[29] A. Passaniti, R. M. Taylor, R. Pili, Y. Guo, P. V. Long, J. A. Haney, R. R. Pauly, D. S. Grant and G. R. Martin, “A Simple, Quantitative Method For Assessing Angiogenesis and Antiangiogenic Agents Using Reconstituted Basement Membrane, Heparin, and Fibroblast Growth Factor,” Laboratory Investigation, Vol. 67, 1992, pp. 519-28.
[30] D. V. Goukassian, G. Qin, C. Dolan, T. Murayama, M. Silver, C. Curry, E. Eaton, C. Luedemann, H. Ma, T. Asahara, V. Zak, S. Mehta, A. Burg, T. Thorne, R. Kishore and D. W. Losordo, “Tumor Necrosis Factor-α Receptor P75 is Required in Ischemia-Induced Neovascularization,” Circulation, Vol. 115, 2007, pp. 752-762.
[31] C. Granet, W. Maslinski and P. Miossec, “Increased AP-1 and NF-κB Activation and Recruitment with the Combination of the Proinflammatory Cytokines IL-17, Tumor Necrosis Factor Alpha and IL-17 in Rheumatoid Synoviocytes,” Arthritis Research Therapy, Vol. 6, 2004, pp. R190-R198.
[32] S. Kobayashi, M. Fukuta, M. Suzuki, H. Tsuneki and I. Kimura, “Inhibitory Effect of Nifedipine on Tumor Necrosis Factor ?-Induced Neovascularization in Cultured Choroidal Explants of Streptozotocin-Diabetic Rat,” Biological and Pharmaceutical Bulletin, Vol. 28, 2005, pp. 242-246.
[33] M. Nomura, W. Ma, N. Chen, A. M. Bode and Z. Dong, “Inhibition of 12-O-Tetradecanoyl Phorbol-13-Acetate- Induced NF-Kappab Activation by Tea Polyphenols, (-)- Epigallocatechin Gallate and Theaflavins,” Carcinogenesis, Vol. 21, 2000, pp 1885-1890.
[34] D. Ichimatsu, M. Nomura, S. Nakamura, S. Moritani, K. Yokogawa, S. Kobayashi, T. Nishioka, and K. Miyamoto, “Structure-Activity Relationship of Flavonoids for Inhibition of Epidermal Growth Factor-Induced Transformation of JB6 Cl 41 Cells,” Molecular Carcinogenesis, Vol. 46, 2007. pp. 436-445.
[35] R. Schreck, B. Meier, D. N. Mannel, W. Droge and P. A. Baeuerle, “Dithiocarbamates as Potent Inhibitors of Nuclear Factor kB Activation in Intact Cells,” Journal of Experimental Medicine, Vol. 175, pp. 1181-1191.
[36] H. Zeitler, Y. Ko, B. Glodny, G. Totzke, M. Appenheimer, A. Sachinidis and H. Vetter, “Cell-Cycle Arrest in G0/ G1 Phase of Growth Factor-Induced Endothelial Cell Proliferation by Various Calcium Channel Blockers,” Cancer Detection and Prevention, Vol. 21, 1997, pp. 332-339.
[37] R. Berkels, G. Egink, T. A. Marsen, H. Bartels, R. Roesen and W. Klaus, “Nifedipine Increases Endothelial Nitric Oxide Bioavailability by Antioxidative Mechanisms,” Hypertension, Vol. 37, 2001, pp. 240-245.
[38] J. P. Cooke, “NO and Angiogenesis,” Atherosclerosis Supplement, Vol.4, 2003, pp. 53-60.
[39] S. Yoshida, M. Ono, T. Shono, H. Izumi, T. Ishbashi, H. Suzuki and M. Kuwano, “Involvement of Interleukin-8, Vascular Endothelial Growth Factor, and Basic Fibroblast Growth Factor in Tumor Necrosis Factor-Alpha- Dependent Angiogenesis,” Molecular and Cellular Biology. Vol. 17, 1997, pp. 4015-4023.
[40] Z Zhou, M. C. Connell and D. J. MacEwan, “TNFR1- Induced NF-?B, but Not ERK, P38mapk or JNK Activation, Mediates TNF-Induced ICAM-1 and VCAM-1 Expression on Endothelial Cell,” Cellular Signaling, Vol. 19, 2007, pp. 1238-1248.
[41] J. Yu, S. Tian, L. Metheny-Barlow, L. J. Chew, A. J. Hayes, H. Pan, G. L. Yu and L. Y. Li, “Modulation of Endothelial Cell Growth Arrest and Apoptosis by Vascular Endothelial Growth Inhibitor,” Circulation Research, Vol. 89, 2001, pp. 1161-1167.
[42] K. Alitalo, T. Tammela and T. V. Petrova, “Lymphangiogenesis in Development and Human Disease,” Nature, Vol. 438, 2005, pp. 946-953.

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