Effects of Asiaticoside on Activation of Nuclear Factor kappa B and Inflammation in RAW264. 7 Cells Induced by Lipopolysaccharide_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

JIANG Chengcheng ¹ , ZHAO Hengguang ² , WU Yamei. ³

Department of Respiratory Medicine, The First People’s Hospital of Chongqing. Chongqing, 400010, ChinaCorresponding Author: ZHAO Heng-guang, E-mail: zhgvip@ 163. com;

Corresponding author：

Keywords：

Asiaticoside; Nuclear factor kappa B; Tumor necrosis factor-α; Interleukin-1; Interleukin-10

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Objective To explore the effects of asiaticoside on the activation of nuclear factor kappa B ( NF-κB) and cytokines expression in RAW264. 7 cells induced by lipopolysaccharide ( LPS) . Methods RAW264. 7 cells were allocated to 5 groups, ie. a blank group, a model group stimulated by LPS at dose of 10 μg/mL, and three asiaticoside treatment groups stimulated by LPS and different doses of asiaticoside simultaneously. The effects of asiaticoside ( 10 - 7 , 10 - 6 , 10 - 5 mol /mL) on the proliferation of cells were examined by MTT assay. The activation of NF-κB was detected and analyzed by the laser scanning confocal microscope( LSCM) ,meanwhile the concentrations of TNF-α, IL-1, and IL-10 in supernatants were quantified by ELISA. Results MTT assay showed that asiaticoside ( 10 - 7 , 10 - 6 ,10 - 5 mol /mL) had no effects on the proliferation of RAW264. 7 cells. Asiaticoside significantly decreased the activation of NF-κB, downregulated the secretion of TNF-αand IL-1, and upregulated IL-10 secretion in a dose dependent manner. According to LSCM, the ratio of NF-κB activation was ( 3. 5 ±1. 5) % , ( 75. 7 ±9. 1) % , ( 66. 8 ±7. 1) % , ( 58. 9 ±9. 0) % , and ( 40. 1 ±8. 8) % in the blank, model, and asiaticoside( 10 - 7 , 10 - 6 , 10 - 5 mol /mL) treatment groups respectively. The contents of TNF-α in supernatants were ( 171. 12 ±35. 42, 1775. 45 ±193. 97,1284. 63 ±162. 13,1035. 22 ±187. 97, 598. 90 ±107. 73) pg/mL respectively and IL-1 were ( 5. 66 ±0. 98,
26. 93 ±3. 48,22. 41 ±2. 84, 17. 05 ±1. 70, 10. 64 ±1. 29) ng/mL respectively, while IL-10 were ( 25. 23 ±2. 17,71. 75 ±8. 31, 82. 82 ±6. 00, 98. 70 ±8. 84, 119. 97 ±9. 13) pg/mL respectively. Conclusion The antiinflammation mechanism of asiaticoside may be mediated by downregulating inflammatory factors through
NF-κB signal pathway and keeping the balance between proinflammatory and antiinflammatory system.

Citation： JIANG Chengcheng,ZHAO Hengguang,WU Yamei.. Effects of Asiaticoside on Activation of Nuclear Factor kappa B and Inflammation in RAW264. 7 Cells Induced by Lipopolysaccharide. Chinese Journal of Respiratory and Critical Care Medicine, 2010, 9(4): 422-425. doi: Copy

1.	刘志峰, 赵慧男, 聂绍良. 积雪草苷药理作用及其机制的研究进展. 广东医学, 2009, 30: 649 -651.
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4.	Kimura Y, Sumiyoshi M, Samukawa K, et al Facilitating action of asiaticoside at low doses on burn wound repair and its mechanism.Eur J Pharmacol, 2008, 28 , 584: 415-423.
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6.	Zhang G, Ghosh S. Molecular mechanisms of NF-kappaB activation induced by bacterial lipopolysaccharide through Toll-like receptors.J Endotoxin Res, 2000, 6: 453-457.
7.	Giuliani C, Napolitano G, Bucci I, et al. NF-κB transcription factor: role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications. Clin Ter, 2001, 152 : 249-253.
8.	鲍国强, 章扬培. NF-κB 的生物学功能. 国外医学分子生物学分册, 2003, 25: 345-348 .
9.	Hawrylowicz CM. Regulatory T cells and IL-10 in allergic inflammation. J Exp Med, 2005, 202 : 1459 -1463 .
10.	Hamilton TA, Ohmori Y, Tebo J. Regulation of chemokine expression by antiinflammatory cytokines. Immunol Res, 2002, 25 :229-245.

1. 刘志峰, 赵慧男, 聂绍良. 积雪草苷药理作用及其机制的研究进展. 广东医学, 2009, 30: 649 -651.
2. Ju-Lin X, Shao-Hai Q, Tian-Zeng L, et al. Effect of asiaticoside on hypertrophic scar in the rabbit ear model. JCutan Pathol, 2009, 36 :234-239.
3. Zhang L, Li HZ, Gong X, et al. Protective effects of Asiaticoside on acute liver injury induced by lipopolysaccharide/D-galactosamine in mice. Phytomedicine, 2010( Epub ahead of print) .
4. Kimura Y, Sumiyoshi M, Samukawa K, et al Facilitating action of asiaticoside at low doses on burn wound repair and its mechanism.Eur J Pharmacol, 2008, 28 , 584: 415-423.
5. 明志君, 孙萌. 积雪草总甙抗炎作用的实验研究. 中国中医药科技, 2002, 9: 62.
6. Zhang G, Ghosh S. Molecular mechanisms of NF-kappaB activation induced by bacterial lipopolysaccharide through Toll-like receptors.J Endotoxin Res, 2000, 6: 453-457.
7. Giuliani C, Napolitano G, Bucci I, et al. NF-κB transcription factor: role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications. Clin Ter, 2001, 152 : 249-253.
8. 鲍国强, 章扬培. NF-κB 的生物学功能. 国外医学分子生物学分册, 2003, 25: 345-348 .
9. Hawrylowicz CM. Regulatory T cells and IL-10 in allergic inflammation. J Exp Med, 2005, 202 : 1459 -1463 .
10. Hamilton TA, Ohmori Y, Tebo J. Regulation of chemokine expression by antiinflammatory cytokines. Immunol Res, 2002, 25 :229-245.

Chinese Journal of Respiratory and Critical Care Medicine

Effects of Asiaticoside on Activation of Nuclear Factor kappa B and Inflammation in RAW264. 7 Cells Induced by Lipopolysaccharide

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