Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide

Jing-hua Peng; Yi-yang Hu; Qin Feng; Yang Cheng; Li-li Xu; Shao-dong Chen; Qing Tao; Feng-hua Li

doi:10.1007/s11655-009-9001-x

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Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide

Updated：2021-08-27

- Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide
- Chinese Journal of Integrative Medicine Vol. 15, Issue 3, Pages: 198-203(2009)
- Affiliations：
  
  Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine,Shanghai,China
- Author bio：
- Funds：
  
  Supported by National Natural Science Foundation of China (No. 30371818);Shanghai Leading Academic Discipline Project (Y0302);Shanghai Educational Development Foundation, (No. 2007CG56);and Shanghai Rising-Star Program (No. 07QA14052)
- DOI：10.1007/s11655-009-9001-x
  CLC：
- Published：2009，
  
  Published Online：26 February 2009，
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Peng, Jh., Hu, Yy., Feng, Q. et al. Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide., Chin. J. Integr. Med. 15, 198–203 (2009). https://doi.org/10.1007/s11655-009-9001-x

Jing-hua Peng, Yi-yang Hu, Qin Feng, et al. Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide. [J]. Chinese Journal of Integrative Medicine 15(3):198-203(2009)
Peng, Jh., Hu, Yy., Feng, Q. et al. Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide., Chin. J. Integr. Med. 15, 198–203 (2009). https://doi.org/10.1007/s11655-009-9001-x DOI：

Jing-hua Peng, Yi-yang Hu, Qin Feng, et al. Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide. [J]. Chinese Journal of Integrative Medicine 15(3):198-203(2009) DOI： 10.1007/s11655-009-9001-x.

摘要

To evaluate the effect of Jianpi Huoxue decoction (健脾活血方

JHD)-containing serum on tumor necrosis factor-α (TNF-α) secretion and endotoxin receptor gene expression in RAW264.7 cells induced by lipopolysaccharide (LPS). The cytotoxicity of blank-control serum and JHD-containing serum at different concentrations were evaluated through the lactate dehydrogenase (LDH) assay in RAW264.7 cells. RAW264.7 cells were divided into six groups: 5% blank-control serum group (C1

n=3)

5% blank-control serum plus LPS group (L1

n=4)

5% JHD-containing serum plus LPS group (J1

n=4)

10% blank-control serum group (C2

n=3)

10% blank-control serum plus LPS group (L2

n=4)

and 10% JHD-containing serum plus LPS group (J2

n=4). After cultured with the corresponding serum for 1 h

cells in L1

J1 and J2 were treated with LPS (0.1 μ g/mL) for 12 h without rinse. The supernate

cells

protein and RNA were collected for assay. TNF-α in the culture supernate was assayed by the enzyme linked immunosorbent assay (ELISA). Protein expression of TNF-α in RAW cells was detected by Western-blot. TNF-α

Toll-like receptor 2 (TLR2)

TLR4 and CD14 mRNA expression in RAW cells were detected by real-time RT-PCR. The LDH assay supported that cultured for 24 h or less with the JHD-containing serum at the concentration of 10% or lower

RAW264.7 cells showed no cytotoxicity. After stimulation with LPS for 2 h

TNF-α in the culture supernate of the 5% blank-control serum plus LPS group (L1

P=0.03)

10% blank-control serum plus LPS group (L2

P=0.002) and in the cell layer (P=0.01) of these groups increased remarkably. After stimulation with LPS for 1 h

the mRNA expression of TNF-α (P=0.004)

TLR (P=0.03)

CD14 (P=0.004) was up-regulated obviously. In the 10% JHD-containing serum plus LPS group (J2)

the protein expression of TNF-α in both supernate (P=0.04) and cell layer (P=0.04)

gene expression of TNF-α (P=0.03)

TLR4 (P=0.001)

CD14 (P=0.001) were all inhibited. On the other hand

the TLR2 mRNA expression was not up-regulated after LPS stimulation in the 10% blank-control serum plus LPS group (L2). JHD-containing serum inhibited the LPS-induced cytokines expression in RAW264.7 which was probably associated with its inhibitory effect on the mRNA expression of LPS receptors TLR and CD14.

Abstract

To evaluate the effect of Jianpi Huoxue decoction (健脾活血方

n=3)

5% blank-control serum plus LPS group (L1

n=4)

5% JHD-containing serum plus LPS group (J1

n=4)

10% blank-control serum group (C2

n=3)

10% blank-control serum plus LPS group (L2

n=4)

and 10% JHD-containing serum plus LPS group (J2

n=4). After cultured with the corresponding serum for 1 h

cells in L1

J1 and J2 were treated with LPS (0.1 μ g/mL) for 12 h without rinse. The supernate

cells

Toll-like receptor 2 (TLR2)

TLR4 and CD14 mRNA expression in RAW cells were detected by real-time RT-PCR. The LDH assay supported that cultured for 24 h or less with the JHD-containing serum at the concentration of 10% or lower

RAW264.7 cells showed no cytotoxicity. After stimulation with LPS for 2 h

TNF-α in the culture supernate of the 5% blank-control serum plus LPS group (L1

P=0.03)

10% blank-control serum plus LPS group (L2

P=0.002) and in the cell layer (P=0.01) of these groups increased remarkably. After stimulation with LPS for 1 h

the mRNA expression of TNF-α (P=0.004)

TLR (P=0.03)

CD14 (P=0.004) was up-regulated obviously. In the 10% JHD-containing serum plus LPS group (J2)

the protein expression of TNF-α in both supernate (P=0.04) and cell layer (P=0.04)

gene expression of TNF-α (P=0.03)

TLR4 (P=0.001)

CD14 (P=0.001) were all inhibited. On the other hand

关键词

Jianpi Huoxue decoction-containing serumlipopolysaccharidecytokineendotoxin receptor

Keywords

Jianpi Huoxue decoction-containing serumlipopolysaccharidecytokineendotoxin receptor

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