侧脑室注射瘦素对OEP大鼠子宫腔液MMP-7蛋白的调节作用

REGULATING EFFECTS OF INTRAVENTRICULAR INJECTION OF LEPTIN ON MMP-7 PROTEIN IN UTERINE FLUID OF OEPRATS

目的:观察侧脑室微量注射瘦素(leptin)对雌性OEP大鼠子宫腔液基质金属蛋白酶-7(MMP-7)的影响。方法:建立卵巢摘除给予雌激素大鼠(OEP)模型,随机分为瘦素注射组(大鼠侧脑室注射瘦素4μl)及生理盐水注射组(大鼠侧脑室注射生理盐水4μl),每组8只大鼠。侧脑室微量注射6h后,提取两组大鼠子宫腔液;采用双向凝胶电泳(2-DE)法分离子宫腔液蛋白,并对其中差异表达蛋白进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术鉴定;最后采用蛋白印迹法比较两组大鼠子宫腔液MMP-7蛋白的表达情况,验证MALDI-TOF-MS的结果。结果:质谱法鉴定经双向凝胶电泳分离的差异目标蛋白为MMP-7;蛋白印迹法显示,瘦素注射组OEP大鼠子宫腔液MMP-7蛋白的表达明显高于生理盐水注射组(P<0.05)。结论:侧脑室微量注射瘦素可在一定时间内对OEP大鼠子宫腔液MMP-7蛋白的表达起到正向调节的作用。

Objective:To observe the effects of intraventricular microinjection of leptin on matrix metalloproteinases-7(MMP-7)in uterine fluid of OEP rats.Methods:OEP rats model was divided into leptin injection group and normal saline injection group with 8 rats in each group.4μl leptin and 4μl normal saline were injected into the lateral ventricle of rats in the 2 groups respectively;and 6 hours after injection,the uterine fluid of rats were obtained.Two-dimensional gel electrophoresis and MALDI-TOF-MS were used to separate and identify the differential proteins in uterine fluid;At last,Western blot was used to compare the MMP-7 expression in uterine fluid of rats in leptin injection group and normal saline injection group to verify the results of MALDI-TOF-MS.Results:The differential target protein separating by two-dimensional gel electrophoresis was MMP-7 identified by MALDI-TOF-MS.Western blot showed that the MMP-7 expression in uterine fluid of rats in leptin injection group was obviously higher than normal saline injection group(P<0.05).Conclusions:Microinjection of leptin into lateral ventricle can positively regulate the MMP-7 protein expression in uterine fiuid of OEP rats for a certain amount of time.