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Journal of Chinese Integrative Medicine ›› 2006, Vol. 4 ›› Issue (5): 509-513.doi: 10.3736/jcim20060514

• Original Experimental Research • Previous Articles     Next Articles

Effects of Qinggan Huoxue Recipe and its decomposed formulas on CD14, Toll like receptor 4 and nuclear factor-κB expressed by Kupffer cells

Guang Ji, Miao Wang, Pei-yong Zheng, Lian-jun Xing, Tao Liu   

  1. Laboratory of Fatty Liver Diseases, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Online:2006-09-30 Published:2006-09-15

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Objective

To investigate the effects of Qinggan Huoxue Recipe (QGHXR), a compound Chinese herbal medicine, and its decomposed formulas Qinggan Recipe (QGR) and Huoxue Recipe (HXR) on expressions of CD14, Toll like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) in Kupffer cells.

Methods

The isolated primary rat Kupffer cells were treated with lipopolysaccharide (LPS) for a certain period of time, a series of concentrations of drug-containing serums of QGHXR and its decomposed formulas were added, the expressions of NF-κB, tumor necrosis factor-α (TNF-α), CD14, and TLR4 of the Kupffer cells were detected in different culture conditions by using Western blot, real-time reverse transcription polymerase chain reaction (RT-PCR) and ELISA methods respectively.

Results

QGR could down-regulate the expression of membrane receptor CD14, but the expression of NF-κB and TNF-α were not significantly decreased after QGR treatment. HXR could down-regulate the expression of membrane receptor TLR4 and inhibit the expressions of NF-κB and TNF-α. QGHXR could down regulate the expressions of membrane receptors CD14 and TLR4 and inhibit the expressions of NF-κB and TNF-α.

Conclusion

QGHXR can protect liver cells by down regulating the expressions of CD14, TLR4 and NF-κB and inhibiting TNF-α expression.

Key words: Kupffer cells, Signal transduction, Qinggan Huoxue Recipe, Toll like receptor 4

Figure 1

Separated KC for 24 hours (Trypan blue staining,×100)"

Figure 2

Latex beads swallowed by KC (Trypan blue staining,×100)"

Table 1

Effects of different doses of LPS on TNF-α expression of KC（ヌ±S , pg/g）"

Group n Emendated TNF-α
Normal control 6 114.99±14.59
10 μg/L 6 134.12±22.86△△
50 μg/L 6 220.77±26.74△△
100 μg/L 6 325.83±60.46**
1 mg/L 6 333.79±60.66**
5 mg/L 6 273.77±46.47**

Table 2

Effects of different LPS cultivated time on TNF-α expression of KC（ヌ±S , pg/g）"

Group n Emendated TNF-α
Normal control 6 261.21±1.44
15 min 6 279.73±1.48△△
30 min 6 303.97±10.68△△
1 h 6 287.07±1.89△△
2 h 6 656.62±10.24**
3 h 6 759.80±13.93**
6 h 6 948.19±6.75**
12 h 6 591.56±9.42**

Figure 3

CD14 expression of KC in 5 groups detected by Western blotting, half-quantity*P<0.05, **P<0.01, vs untreated group, n=3."

Figure 4

TLR4 expression of KC in 5 groups detected by Western blotting, half-quantity*P<0.05, **P<0.01, vs untreated group, n=3."

Figure 5

NF-κB expression of KC in 5 groups detected by Western blotting, half-quantity*P<0.05, **P<0.01, vs untreated group, n=3."

Figure 6

Effects of drug-containing serums on TNF-α mRNA expression of KC in 5 groups detected by RT-PCR, half-quantity*P<0.05, **P<0.01, vs untreated group, n=3."

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