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重金属铅污染对兔膝关节软骨中TIMP-1和MMP-13及滑膜中TNF-α和IL-1β含量的影响

Expression of TIMP-1 and MMP-13 in Rabbit Articular Cartilage of Knee Joint or Expression of TNF-α and IL-1β in Synovial to Heavy Metal Lead Exposure

【作者】 刘云忠

【导师】 毕树雄;

【作者基本信息】 山西医科大学 , 外科学, 2011, 硕士

【摘要】 目的:1.观察饮水型重金属铅污染时,其在新西兰兔的血液、骨骼、膝关节软骨及滑膜中含量的分布情况;2.分析饮水型重金属铅污染对新西兰兔膝关节软骨TIMP-1和MMP-13及滑膜TNF-α和IL-1βmRNA表达的影响。方法:取34只2.5月龄的新西兰兔,雌雄不限,体重1.3-1.5Kg。按饲养期间饮水质量不同,随机分为两组进行饲养,每组17只。对照组:饮用正常水;实验组:饮用1.0 g/L醋酸铅水。连续饲养3个月后,采用耳缘静脉空气栓塞处死,分别取其血液、骨骼、关节软骨及滑膜,检测其中的重金属铅含量。同时剥取膝关节面软骨约200mg置冻凝管,液氮冷冻保存,提取软骨和滑膜中的mRNA,并反转录和扩增,用实时荧光定量反转录聚合酶链式反应(Real time RT-PCR)法测定兔膝关节软骨中TIMP-1、MMP-13及滑膜中TNF-α、IL-1β的mRNA表达。结果:1.对照组和实验组的血液、骨骼、关节软骨及滑膜中都可检测到重金属铅,两组中均以骨骼中含量最高,依次为滑膜、软骨和血液。与对照组相比,实验组中血液、骨骼、关节软骨及滑膜中铅的含量,均明显增加(P<0.05)。2.实验组膝关节软骨中TIMP-1和MMP-13的mRNA表达量,较对照组,其TIMP-1明显减少,而MMP-13明显增多;实验组滑膜中TNF-α和IL-1β的mRNA表达量,均较对照组明显增多(P<0.05)。结论:通过连续3个月的饮用含铅水染毒,在新西兰兔血液、骨骼、膝关节软骨及滑膜中均可检测到重金属铅,并且其对关节软骨和滑膜均有明显损害作用。

【Abstract】 Object:1. To study the blood, bone and articular cartilage and synovial of lead content in the rabbits which was drinked lead pollution of water2. To study the mRNA of TIMP-1 and MMP-13 in the knee joint articular cartilage and the mRNA of TNF-αand IL-1βin the synovial in response to lead pollution of water.Method:At the age of 2.5 months,34 New Zealand rabbits(1.3-1.5Kg) which were divided into 2 parts started drinking lead pollution of water and pure water.The water of concentration which was polluted by lead is 1.0 g/L.Thereafter,the rabbits were raised for 3 months.Then,they will be executed by ear margin veins air embolism, take its blood,bone, cartilage and synovial respectively,and test lead content by machine.At the same time, about 200mg knee cartilage will be restore in tube and frozen in the liquid nitrogen cryopreservation.we extracte the Mrna and measue the mRNA expressions in the knee cartilage and synovial with RT-PCR..Result:1. The control group and lead exposure group’s blood, bone, cartilage and synovial all can be detected , but two groups of heavy metal lead with bones in both the highest levels, which in turn for synovial, cartilage and blood. Compared with control group, the amount of lead in the group of ead exposure were significantly increased in the blood, bone, cartilage and synovial(P < 0.05).2. Compared with control group,the mRNA of TIMP-1 is significantly reduced in the lead exposure group,but the mRNA of MMP-13 is significantly increased.and the mRNA of TNF-αand IL-1βin the synovial is significantly increased(P < 0.05).Conclusion:When the rabbits was drinked for 3 months,we can measure the lead in the blood, bone, knee cartilage and synovial,and it can damage the cartilage and synovial significantly.

【关键词】 铅暴露关节软骨滑膜TIMP-1MMP-13TNF-αIL-1βRT-RCR
【Key words】 Lead exposurArticular cartilageSynovialTIMP-1MMP-13TNF-αIL-1βRT-RCR
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