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Anti-inflammatory Effects of Ethanol Extract from Kummerowia striata (Thunb.) Schindl on LPS-Stimulated RAW 264.7 Cell

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Abstract

Kummerowia striata (Thunb.) Schindl has long been used as a fork herb in inflammation-related therapy. This study was undertaken to determine the anti-inflammatory effect of the plant. High performance liquid chromatography (HPLC) was used for evaluating the extract. While dexamethasone (DM) was used as a positive control, the effects of ethanol extract on the production of IL-1β, IL-6, NO, COX-2 and TNF-α, the expression of iNOS mRNA, TNF-α mRNA, COX-2 mRNA, protein production of COX-2 and HO-1, NF-κB and I-κB of LPS-stimulated RAW 264.7 cells were studied by sandwich ELISA, real-time PCR, Western blot analysis and immunocytochemistry assay respectively. The results showed that K. striata (Thunb.) Schindl had a good anti-inflammatory effect on LPS-stimulated RAW264.7 cell. On one hand, it could significantly inhibit the production of IL-1β, IL-6, NO, TNF-α, COX-2 in LPS-stimulated cell than that of single LPS stimulated cell (p < 0.01 or p < 0.05). On the other hand, it could increase the production of IL-10 and HO-1 than that of single LPS intervention cell (p < 0.01 or p < 0.05). Furthermore, the extract also could inhibit the production of NF-κB and I-κB compared to single LPS stimulated cell. In a word, it suggested that the anti-inflammatory actions of K. striata (Thunb.) Schindl ethanol extract might be due to the down-regulation of IL-1β, IL-6, NO, TNF-α and COX-2 via the suppression of NF-κB activation and conversation of I-κB production, and another pathway was up regulating the production of IL-10 and HO-1.

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Abbreviations

LPS:

lipopolysaccharide

PBS:

phosphate buffered saline

TBST:

Tris.HCl buffered saline Tween

Real-time PCR:

real-time polymerase chain reaction

ELISA:

enzyme-linked immunosorbent assay

iNOS:

inducible nitric oxide syntheses chain reaction

HO-1:

heme oxygenase-1

NF-κB:

nuclear factor-κB

I-κB:

inhibitory factor-κB

IL-1β:

interleukin-1 beta

TNF-α:

tumor necrosis factor-alpha

NO:

nitric oxide

IL-10:

interleukin-10

IL-6:

interleukin-6

COX-2:

cyclooxygenase-2

CPE:

cytopathic effect test

APS:

astragalus polysaccharides

DM:

dexamethasone

HPLC:

high performance liquid chromatography

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Acknowledgement

We would like to thank Prof. Chen Ke-li (Hubei College of Traditional Chinese Medicine) for identification of the plant, Mr. Leng Yi-bing (College of Life Science and Technology, Huazhong University of Science and Technology) for Western blot analysis.

This work was supported by Hubei & Wuhan Engineering Technology Center of Modernization Traditional Chinese Medicine, Wuhan Jianmin Pharmacy of TCM Co., Ltd, Wuhan, 430054, People’s Republic of China.

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Authors and Affiliations

  1. College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, People’s Republic of China

    Jun-Yan Tao & Xiao-Yu Zhang

  2. Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, People’s Republic of China

    Lei Zhao & Shu-Ling Zhang

  3. Hubei and Wuhan Engineering Technology Center of Modernization Traditional Chinese Medicine, Wuhan Jianmin Pharmacy of TCM Co., Ltd, Wuhan, 430054, People’s Republic of China

    Zhi-Jun Huang, Qiong-Guang Zhang,  Fei-Xiao & Bao-Hui Zhang

  4. School of Pharmacy, Hubei College of Traditional Chinese Medicine, Wuhan, 430065, People’s Republic of China

    Guo-Hua Zheng

  5. Pharmaceutical Department, Wuhan Centre Hospital, Wuhan, 430014, People’s Republic of China

    Qi-Lin Feng

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  1. Jun-Yan Tao
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Correspondence to Xiao-Yu Zhang or Guo-Hua Zheng.

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Jun-Yan Tao and Lei Zhao have equally contributed to this work.

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Tao, JY., Zhao, L., Huang, ZJ. et al. Anti-inflammatory Effects of Ethanol Extract from Kummerowia striata (Thunb.) Schindl on LPS-Stimulated RAW 264.7 Cell. Inflammation 31, 154–166 (2008). https://doi.org/10.1007/s10753-008-9061-7

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