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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (7): 636-641.doi: 10.3736/jcim20090707

• Original Experimental Research • Previous Articles     Next Articles

Effects of genistein on interleukin-1β and tumor necrosis factor-α secreted by fibroblast-like synoviocytes isolated from type Ⅱ collagen-induced arthritis rats

Bo Gao, Yu Zhang),Li Ma, Xue-zeng Zhang   

  1. Department of Hematopathy and Rheumatism, Clinical Medicine College,Yangzhou University, Yangzhou 225001, Jiangsu Province, China
  • Received:2009-02-25 Accepted:2009-04-22 Online:2009-07-20 Published:2009-07-15
  • Contact: Yu Zhang E-mail:yzzy10182001@yahoo.com.cn

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Objective

To investigate the effects of genistein (Gen) on interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) secreted by fibroblast-like synoviocytes (FLSs) of rats with type Ⅱ collagen-induced arthritis (CIA).

Methods

Type Ⅱ collagen was injected to induce arthritis in rats, and arthritis index was applied to evaluate whether the arthritis was induced successfully. The primary FLSs were separated from synovial membranes by using type Ⅱ collagenase digestion. Then the expression of vascular cell adhesion molecule-1 (VCAM-1) was estimated by flow cytometry (FCM). After addition of different concentrations of Gen into the FLSs, IL-1β and TNF-α contents in the supernatants were measured by enzyme-linked immunosorbent assay.
Results

Three days after collagen injection, the rats started to develop arthrocele and the arthritis index increased gradually. The arthritis index of CIA group was significantly higher than that of the control group. The expression of VCAM-1 was up to 85.5% in the 4th generation FLSs, indicating that most part of the cultured cells were type-B synoviocytes. After administration of different concentrations of Gen (100, 200, 400 μmol/L), the contents of IL-1β and TNF-α in supernatants of FLSs were decreased dose-dependently.
Conclusion

Genistein can suppress the secretion of TNF-α and IL-1β in FLSs dose-dependently, which may be one of the mechanisms for genistein in inhibiting the arthromeningitis of CIA rats.

Key words: Arthritis, Genistein, Synoviocyte, Interleukin-ip, Tumor necrosis factor-a, Rats

Table 1

Scores of arthritis index of CIA rats ($\bar{x}$±s)"

Group n Score of arthritis index
1st week 2nd week 3rd week 4th week 5th week 6th week
Control 5 0 0 0 0 0 0
CIA 5 5.20±1.48* 7.40±2.30* 8.40±2.40* 7.80±1.30* 7.60±1.67* 7.40±1.34*

Figure 1

Photos of synoviocytes cultured for different time A: After 24-hour culture, the primary synovial cells had the figures of fibroblasts (Light mioroscopy, ×400); B: Colony growth of the fibroblast-like synoviocytes was observed at passage culture (Light mioroscopy, ×100)."

Figure 2

Expression of VCAM-1 in fibroblast-like synoviocytes The green curve represents fibroblast-like synoviocytes stained with PE mouse IgG1, κ isotype control; the red curve represents fibroblast-like synoviocytes stained with PE-conjugated mouse anti-rat VCAM-1."

Table 2

Effects of Gen on IL-1β and TNF-α production from the supernatant of cultured fibroblast-like synoviocytes stimulated by LPS ($\bar{x}$±s)"

Group n Drug concentration TNF-α IL-1β
Control 6 1.005±0.017▲ 1.001±0.038▲
CIA 6 1.780±0.053*▲ 1.405±0.054*
MTX 6 1 mg/L 1.505±0.032△ 1.231±0.044△
Gen1 6 100 μmol/L 1.742±0.039▲ 1.366±0.055▲
Gen2 6 200 μmol/L 1.484±0.041△□ 1.250±0.071△
Gen3 6 400 μmol/L 1.383±0.028△▲□ 1.140±0.048△▲□
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