| Objective: To investigate the effect on the chondrocytes which cultured in vitro withsoy isoflavone. To reveal furtherly the role of estrogen in the process of the articularcartilage cataplasis in osteoarthritis.Methods: 1. 3 female,3-week-old rabbits, articular cartilage were isolated thearticular from the distal and proximal femoral cartilage in asepsis, then digested intochondrocytes. The first generation chondrocytes were randomly divided into 8groups: control group, E2, 10-5mol/L-10-10mol/L SI. Every hole of the 96-hole-plateculture 2×103 chondrocytes,every group include 4 parallel holes; Every hole of the24-hole-plate culture 104 chondrocytes,every group include 3 parallel holes, putted acoverslip which is 5mm×5mm in the bottom of 24-hole-plate.After 48 hourslater,interfered with difference density drugs,changed the culture medium every 3days. MTT method were used to determine the proliferation of cultured cartilagechondrocytes in the presence of different drugs every day, measured continously 8days. After 7 days,dyeing the chondrocytes adhered on the coverslip using toluidineblue staining solution, immunocytochemistry (for ERβ), Wright's staining solution.2. The first generation chondrocytes were randomly divided into 2 groups: Agroup weren't interfered with IL-β1, and B group were interfered with withIL-β1.Every group were randomly divided into 8 subgroups: control group, E2,10-5mol/L SI, 10-6mol/L-10-10mol/L SI. After 48 hours later,A group were interferedwith difference density drugs for 5 days,B group were interfered with 10ng/ml IL-1βfor 24 hours first, then were treated with blank, E2, 10-5mol/L-10-10mol/L SI for 5days, changed the culture medium every 3 days. Total RNA was extracted from thechondrocytes and the expression of typeâ… ,â…¡collagen,Biglycan,TGF-β1,IGF-1genes were investgated by the reverse transcription- polymerase chain reaction(RT-PCR) method.Results: 1.The results of MTT show us: every group are similar in first 24h(p＞0.05);10-5mol/L SI group is low significantly than the control group from the secondday to the eight day(p＜0.05); 10-6mol/L and 10-10mol/L SI group are similar to thecontrol group anytime (p＞0.05); 10-7mmol/L,10-8mol/L and 10-9mol/L SI group are height significantly than the control group from the third day to the eightday(p＜0.05);The immunocytochemistry (for ERβ) show us a positive result,anachromasis brown drops are founded in the endochylema and/or nucleus ofeverygroup; Toluidine blue staining show us amaranth drops in some endochylema ofevery group.2.The result of RT-PCR indicated that every group can express typeâ…¡collagen,bilgcan,TGF-β1 mRNA.The mRNA of collagen,bilgcan,TGF-β1 were inhibited by10-5mol/L SI and IL-1β,and the deprssant effect could be aggravated by 10-5mol/LSI,and be lessened by 10-7mol/L, 10-8mol/L SI. The mRNA of typeâ…¡collagen,bilgcan,TGF-β1 in normal chondrocytes can be increased by10-7mol/L, 10-8mol/L SI. The mRNA of typeâ… collagen could be inducted andexpressed by 10-5mol/L,10-6mol/L,10-7mol/L SI in the normal chondrocytes.Thedegenerative chondrocytes can express the mRNA of typeâ… collagen. The mRNA oftypeâ… collagen could be decreased by 10-8mol/L, 10-9mol/L SI. The mRNA of IGF-1could be inducted and expressed by 10-7mol/L, 10-8mol/L, 10-9mol/L SI in the normalchondrocytes,and be inducted and expressed by 10-7mol/L,10-8mol/L,in thedegenerative chondrocytes.Conclusion: 1. Different density of soy isoflavone have double-effect on the cartilagechondrocytes which cultured in vitro. 10-5mol/L SI could inhibitive to theproliferation, 10-7mol/L, 10-8mol/L,10-9mol/L SI could promote the proliferation, andthe effect could be enhanced with the density of SI be advanced from 10-9mol/L to10-7mol/L; 2. Different density of soy isoflavone have different effect on thechondrocyte to the express of mRNA of collagen, biglycan,IGF-1, TGF-β1. 3.Thecartilage chondrocytes which cultured in vitro can express ERβ, the effect of soyisoflavone on the chondrocyte is like with the estrogen, may be through estrogenacceptor pathway;The time of promote the proliferation of the chondrocytes by10-7mol/L SI, 10-8mol/L SI, 10-9mmol/L SI in vitro is later than the 17β-estrodiol,may be accosiated the drug action of SI be weaker than the 17β-estrodiol. 4.Theproliferation of cartilage chondrocytes can be promoted by soy isoflavone. The effectof soy isoflavone on the collagen and biglycan is better than the estrodiol. These supply a pharmaco- and pharmacodynamics evidence for soy isoflavone whichprevent and cure to osteoarthritis in clinical. 5. The difference of effect between soyisoflavone and estodiol shows: 1) The rates of soy isoflavone combined to estrogenreceptor alpha and beta may be different to estrodiol; 2) The effect pathway tochondrocytes of soy isoflavone is different to estrodiol.
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