不同浓度雌激素对软骨细胞金属蛋白酶mRNA表达的影响

Effect of estrogen on MMPs mRNA expression of chondrocytes

目的 观察不同浓度雌激素对体外培养兔关节软骨细胞金属蛋白酶mRNA表达的影响。方法 体外培养雌兔关节软骨细胞 ,随机分为A、B两组 ,A组中加入 17β 雌二醇 0、10 -6、10 -7、10 -8、10 -9、10 -10 、10 -11、10 -12 mol/L干预 72h ;B组先用 10mg/mlIL 1β干预 2 4h ,随后分别加入 0、10 -6～ 10 -12 mol/L 17β 雌二醇作用 72h。采用RT PCR方法观察软骨细胞金属蛋白酶mRNA表达状况。结果 10 -6、10 -8、10 -9、10 -12 mol/L雌二醇组 ,IL 1β +雌二醇 0mol/L组及IL 1β+雌二醇 10 -11mol/L表达MMP 1mRNA ;正常软骨细胞不表达MMP 8mRNA ,10 -8、10 -9、10 -12mol/L雌二醇组及IL 1β +雌二醇 10 -6、10 -7、10 -10 mol/L组表达MMP 8mRNA ;所有软骨细胞均未表达MMP 13mRNA ;正常软骨细胞不表达TIMP 1mRNA ,10 -6、10 -8、10 -10 、10 -11mol/L雌二醇组及IL 1β +雌二醇 10 -12 mol/L组表达TIMP 1mRNA。结论 雌激素对关节软骨细胞金属蛋白酶表达的调控作用随其浓度变化而不同。低于生理浓度的雌激素 (10 -12 mol/L)对延缓软骨退变较适宜。

Objective To observe the effect of estrogen on MMPs mRNA expression in articular cartilage chondrocytes.Method Chondrocytes isolated from the distal femoral and proximal tibial cartilage of 4 month old female rabbits were divided into 2 groups.One group treated with 0,10 -6 ,10 -7 ～10 -12 mol/L estradiol for 72 hours.The other group incubated with 10 mg/ml IL 1β for 24 hours first,then treated with 0,10 -6 ,10 -7 ～10 -12 mol/L estradiol for 72 hours.Total RNA was extracted from the chondrocytes and the cxpression of typeⅠ,Ⅱ and Ⅲ collagen genes was investigated by the reverse transcription polymerase chain reaction (RT PCR) method.Results There was no expression of MMP 1 in normal chondrocytes,but the expression of MMP 1 was found in 10 -6 ,10 -8 ,10 -9 and 10 -12 mol/L estradiol and IL 1β+10 -11 mol/L estradiol treated chondrocytes;expression of MMP 8 was found in 10 -8 ,10 -9 and 10 -12 mol/L estradiol and IL 1β+10 -6 ,10 -7 and 10 -10 mol/L estradiol treated chondrocytes,but not in normal control.Expression of MMP 13 was not found in all chondrocytes;There was no expression of TIMP 1 in normal chondrocytes,but the expression of TIMP 1 was found in 10 -6 ,10 -8 ,10 -10 and 10 -11 mol/L estradiol and IL 1β +10 -12 mol/L estradiol treated chondrocytes.Conclusion Estrogen can modulate MMPs mRNA expression in articular cartilage chondrocytes.Estrogen concentration lower than physiological level (10 -12 mol/L) can delay the degradation of articular cartilage.