Search JIM Advanced Search

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


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).


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.

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.

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△▲□
[1] Firestein GS . Invasive fibroblast-like synoviocytes in rheumatoid arthritis. Passive responders or transformed aggressors[J]. Arthritis Rheum, 1996,39(11):1781-1790
doi: 10.1002/(ISSN)1529-0131
[2] Feldmann M, Brennan FM, Maini RN . Role of cytokines in rheumatoid arthritis[J]. Annu Rev Immunol, 1996,14(1):397-443
doi: 10.1146/annurev.immunol.14.1.397
[3] Wang SY, Li ZC, Bu YJ . Research advances on biological function of soybean isoflavone[J]. Xinxiang Yi Xue Yuan Xue Bao, 2005,22(7):397-400
王守英, 李志春, 卜勇军 . 大豆异黄酮的生物学作用研究进展[J]. 新乡医学院学报, 2005,22(7):397-400
[4] Barrett JR . The science of soy: what do we really know?[J]. Environ Health Perspect, 2006,114(6):A352-A358
doi: 10.1289/ehp.114-a352 pmid: 1480510
[5] Seki N, Sudo Y, Yoshioka T, Sugihara S, Fujitsu T, Sakuma S, Ogawa T, Hamaoka T, Senoh H, Fujiwara H . Type Ⅱ collagen-induced murine arthritis. Ⅰ. Induction and perpetuation of arthritis require synergy between humoral and cell-mediated immunity[J]. J Immunol, 1988,140(5):1477-1484
[6] Yin G, Liu W, An P, Li P, Ding I, Planelles V, Schwarz EM, Min W . Endostatin gene transfer inhibits joint angiogenesis and pannus formation in inflammatory arthritis[J]. Mol Ther, 2002,5(5 pt 1):547-554
doi: 10.1006/mthe.2002.0590
[7] Sundarrajan M, Boyle DL, Chabaud-Riou M, Hammaker D, Firestein GS . Expression of the MAPK kinases MKK-4 and MKK-7 in rheumatoid arthritis and their role as key regulators of JNK[J]. Arthritis Rheum, 2003,48(9):2450-2460
doi: 10.1002/(ISSN)1529-0131
[8] Jiao K, Zhang SS. The enzyme-linked immunoassay and its application[M]. Beijing: Chemical Industry Press, 2004: 2
焦奎, 张书圣 . 酶联免疫分析技术及应用[M]. 北京: 化学工业出版社, 2004: 2
[9] Guiducci S, Del Rosso A, Cinelli M, Margheri F , D'Alessio S, Fibbi G, Matucci Cerinic M, Del Rosso M .Rheumatoid synovial fibroblasts constitutively express the fibrinolytic pattern of invasive tumor-like cells[J]. Clin Exp Rheumatol, 2005,23(3):364-372
[10] Morales-Ducret J, Wayner E, Elices MJ, Alvaro-Gracia JM, Zvaifler NJ, Firestein GS . Alpha 4/beta 1 integrin(VLA-4) ligands in arthritis. Vascular cell adhesion molecule-1 expression in synovium and on fibroblast-like synoviocytes[J]. J Immunol, 1992,149(4):1424-1431
[11] Fox DA . Cytokine blockade as a new strategy to treat rheumatoid arthritis: inhibition of tumor necrosis factor[J]. Arch Intern Med, 2000,160(4):437-444
doi: 10.1001/archinte.160.4.437
[12] Youn J, Kim HY, Park JH, Hwang SH, Lee SY, Cho CS, Lee SK . Regulation of TNF-alpha-mediated hyperplasia through TNF receptors, TRAFs, and NF-kappaB in synoviocytes obtained from patients with rheumatoid arthritis[J]. Immunol Lett, 2002,83(2):85-93
doi: 10.1016/S0165-2478(02)00079-2
[13] Horai R, Saijo S, Tanioka H, Nakae S, Sudo K, Okahara A, Ikuse T, Asano M, Iwakura Y . Development of chronic inflammatory arthropathy resembling rheumatoid arthritis in interleukin 1 receptor antagonist-deficient mice[J]. J Exp Med, 2000,191(2):313-320
doi: 10.1084/jem.191.2.313
[14] Bresnihan B, Alvaro-Gracia JM, Cobby M, Doherty M, Domljan Z, Emery P, Nuki G, Pavelka K, Rau R, Rozman B, Watt I, Williams B, Aitchison R , McCabe D, Musikic P .Treatment of rheumatoid arthritis with recombinant human interleukin-1 receptor antagonist[J]. Arthritis Rheum, 1998,41(12):2196-2204
doi: 10.1002/(ISSN)1529-0131
[15] Zhu GC, Ding Z, Chen ZS, Dong C, Guo H, Chen BC . Experimental study on genistein prevention and treatment of transplant arteriosclerosis in aortic transplants of rat[J]. Transplant Proc, 2006,38(10):3307-3308
doi: 10.1016/j.transproceed.2006.10.114
[16] Zhao JH, Arao Y, Sun SJ, Kikuchi A, Kayama F . Oral administration of soy-derived genistin suppresses lipopolysaccharide-induced acute liver inflammation but does not induce thymic atrophy in the rat[J]. Life Sci, 2006,78(8):812-819
doi: 10.1016/j.lfs.2005.05.104
[17] Davis JN, Kucuk O, Sarkar FH . Genistein inhibits NF-kappa B activation in prostate cancer cells[J]. Nutr Cancer, 1999,35(2):167-174
doi: 10.1207/S15327914NC352_11
[1] Chiung-Hui Huang, Mei-Ling Yeh, Fang-Pey Chen, Daphne Wu. Low-level laser acupuncture reduces postoperative pain and morphine consumption in older patients with total knee arthroplasty: A randomized placebo-controlled trial. Journal of Integrative Medicine, 2022, 20(4): 321-328.
[2] Ye-juan Jia, Tian-yu Li, Peng Han, Yu Chen, Li-jia Pan, Chun-sheng Jia. Effects of different courses of moxibustion treatment on intestinal flora and inflammation of a rat model of knee osteoarthritis. Journal of Integrative Medicine, 2022, 20(2): 173-181.
[3] Ju-Yeon Park, Young-Won Kwon, Sun-Ah Kim, Sun-Dong Park, Chang-Hyun Kim, Jin-Hee Kim, Ju-Hee Lee. Polyherbal formula SC-E3 inhibits rheumatoid arthritis activity in a mouse model of type-II collagen-induced arthritis. Journal of Integrative Medicine, 2021, 19(3): 265-273.
[4] Bing-rong Li, Shi-yun Shao, Long Yuan, Ru Jia, Jian Sun, Qing Ji, Hua Sui, Li-hong Zhou, Yi Zhang, Hui Liu, Qi Li, Yan Wang, Bi-meng Zhang. Effects of mild moxibustion on intestinal microbiome and NLRP3 inflammasome in rats with 5-fluorouracil-induced intestinal mucositis. Journal of Integrative Medicine, 2021, 19(2): 144-157.
[5] Johannes Loeser, Brigitte Layer, Christopher Plata, Klaus-Maria Perrar, Tim Hucho, Rebecca Kulbida. Hirudotherapy attenuates arthritic pain in patients with various chronic pain syndromes: A retrospective analysis. Journal of Integrative Medicine, 2020, 18(5): 425-433.
[6] Sitthichai Iamsaard, Supatcharee Arun, Jaturon Burawat, Supataechasit Yannasithinon, Saranya Tongpan, Sudtida Bunsueb, Natthapol Lapyuneyong, Pannawat Choowong-in, Nareelak Tangsrisakda, Chadaporn Chaimontri, Wannisa Sukhorum. Evaluation of antioxidant capacity and reproductive toxicity of aqueous extract of Thai Mucuna pruriens seeds. Journal of Integrative Medicine, 2020, 18(3): 265-273.
[7] Sandip B. Bharate, Vikas Kumar, Sonali S. Bharate, Bikarma Singh, Gurdarshan Singh, Amarinder Singh, Mehak Gupta, DeepikaSingh, AjayKumar, Surjeet Singh, Ram A. Vishwakarma. Discovery and preclinical development of IIIM-160, a Bergenia ciliata-based anti-inflammatory and anti-arthritic botanical drug candidate. Journal of Integrative Medicine, 2019, 17(3): 192-204.
[8] Morufu Eyitayo Balogun, Elizabeth Enohnyaket Besong, Jacinta Nkechi Obimma, Ogochukwu Sophia Mbamalu, Fankou Serges Athanase Djobissie. Protective roles of Vigna subterranea (Bambara nut) in rats with aspirin-induced gastric mucosal injury. Journal of Integrative Medicine, 2018, 16(5): 342-349.
[9] Lucky Legbosi Nwidu, Raphael Ellis Teme. Hot aqueous leaf extract of Lasianthera africana (Icacinaceae) attenuates rifampicin-isoniazid-induced hepatotoxicity. Journal of Integrative Medicine, 2018, 16(4): 263-272.
[10] Katy Andrade Monteiro Zacaron, Nélia da Silva Mendes, Yuri Cotta e Silva, Giancarlo Lucchetti. Effects of laying on of hands with and without a spiritual context on pain and functionality of older women with knee osteoarthritis: Study protocol for a randomized controlled trial. Journal of Integrative Medicine, 2018, 16(2): 106-112.
[11] Kyeong Mee Park, Tae Hwan Cho. Therapeutic effect of acupuncture point injection with placental extract in knee osteoarthritis. Journal of Integrative Medicine, 2017, 15(2): 135-141.
[12] Susana Seca, Sebastian Kirch, António S.Cabrita, Henry J.Greten. Evaluation of the effect of acupuncture on hand pain, functional deficits and health-related quality of life in patients with rheumatoid arthritis—A study protocol for a multicenter, double-blind, randomized clinical trial. Journal of Integrative Medicine, 2016, 14(3): 219-227.
[13] MesfinYimam, Young-Chul Lee, Breanna Moore, Ping Jiao, MeiHong, Jeong-Bum Nam, Mi-Ran Kim, Eu-Jin Hyun, Min Chu, Lidia Brownell, Qi Jia. Analgesic and anti-inflammatory effects of UP1304, a botanical composite containing standardized extracts of Curcuma longa and Morus alba. Journal of Integrative Medicine, 2016, 14(1): 60-68.
[14] Kylie Connolly, Douglas Jackson, Candice Pullen, Andrew Fenning. Alpha-adrenoceptor antagonism by Crassostrea gigas oyster extract inhibits noradrenaline-induced vascular contraction in Wistar rats. Journal of Integrative Medicine, 2015, 13(3): 194-200.
[15] Udhaya Lavinya Baskaran, Sherry Joseph Martin, Rasool Mahaboobkhan, Sabina Evan Prince. Protective role of Triphala, an Indian traditional herbal formulation, against the nephrotoxic effects of bromobenzene in Wistar albino rats. Journal of Integrative Medicine, 2015, 13(2): 115-121.
Full text



[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278