强力霉素对退变腰椎间盘中MMP-3表达水平的调控

Doxycycline to degeneration of lumbar MMP-3 expression level of control

目的研究强力霉素对退变腰椎间盘中基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)表达水平的调控作用。方法 (1)按随机数字表随机将40只大鼠分为对照组和实验组各20只,制备动物模型,8周后实验组动物每日皮下注射强力霉素25 mg,分早晚两次注射,对照组动物每日皮下注射等体积的生理盐水,分早晚两次注射。2周后取腰椎间盘组织,一部分放置10%中性福尔马林固定,另一部分存放在液氮中。(2)免疫组织化学检测:取福尔马林固定好的样本应用免疫组织化学SP法检测腰椎间盘组织中MMP-3的表达,阳性表达为为棕黄色,主要存在于细胞浆内。(3)Western-bloting检测:用液氮中保存的样品采用抽提椎间盘髓核的蛋白,转膜后X线显影,测平均相对含量。结果实验组大鼠椎间盘组织MMP～3表达低于对照组(P<0.05)。结论新型MMPs抑制剂强力霉素能降低退变腰椎间盘MMP-3表达水平。

Objective To study the regulative function of Doxycycline on the expression level of MMP-3 in degenerated lumbar intervertebral disc.Methods （1）To divide 40 rats into an experimental group and a contrastive group stochastically by random number table, each group 20 rats.Then complete the animal model. From the 8th week after operation,the animals of the experimental group were injected with Doxycycline 25mg,two times in the daytime and in the night per day.Those of the control group with the equal volume of saline per day.Two weeks late,the tissue of degenerated lumbar intervertebral disc was taken,A part was put into 10% neutral formalin to be fixed,another part was deposited in the liquid nitrogen.（2）Immumo-histochemistry Test.To Take the sample which was fixed by formalin,then to test the expression of MMP-3 in degenerated lumbar intervertebral disc by the way of SP of immumo-histochemistry.The positive part expression are yellowish brown, which mainly exists in the cytoplasma.（3）Western-bloting Test. The sample,which was in liquid nitrogen, was extracted the vertebral pulp protein in lumbar intervertebral disc,then use X development to measure the average relative content of protein.Results The expression of MMP-3 in the experimental group decreased and significantly different from the control group （P〈0.05） .Conclusion The new MMFs inhibitor-Doxycycline can decrease expression level of MMP-3 lumbar intervertebral disc.