磨损颗粒诱导细胞外基质金属蛋白酶诱导因子表达及机制研究

A study of extracellular matrix metalloproteinaseinducer expression induced by wear particles

[目的]观察磨损颗粒诱导EMMPRIN产生及PDTC对EMMPRIN表达影响,探讨EMMPRIN在炎症反应中的作用及其调节机制。[方法]构建小鼠air-pouch动物模型,实验组囊腔内注射颗粒悬液3 ml(B、C组),对照组注射磷酸盐缓冲盐水3 ml(A组),即日起A组、C组每天腹腔注射0.1 ml PDTC溶液,B组注射等量生理盐水,给药后第3、7、14 d后各处死动物8只取出air-pouch囊壁组织,行大体、组织学观察,半定量RT-PCR、免疫组织化学、western-blot检测。[结果]在B组、C组小鼠背部可见一白色、质脆椭圆形囊壁组织,光镜下可见大量炎性细胞浸润,A组背部皮下可见一透明壁薄囊腔,磨损颗粒均能诱导囊壁组织EMMPRINmRNA及蛋白的表达,且呈时间依赖性。7、14 d C组EMMPRIN mRNA、蛋白及NF-κBP65的表达少于B组(P<0.05),A组明显少于C组(P<0.01)。[结论]磨损颗粒可以刺激EMMPRIN的产生,EMMPRIN的生成可能受NF-κB的调节。

[Objective] To observe the expression of extracellular matrix metalloproteinase inducer （EMMPRIN） and its regulatory mechanism under the influence of pyrrolidine dithiocarbamate （ PDTC）, when induced by wear debris. [ Methods ] Using the murine air pouch model of inflammation, 24 Kunming mice were divided into 3 groups （8 per group）, A, B, and C, for this study. The air pouches of the mice in groups B and C were injected with a 3 ml suspension containing 1×10^8/ml poly- ethylene particles and the pouchesof the mice in group A were injected with 3 ml saline. The mice in groups A and C were injec- ted with 0. 1 ml PDTC solution in the stomach and the mice in group B were injected with saline. The animals were sacrificed 3d, 7d, and 14 d after the injection. The histological changes were assessed usinga light microscope. The gene and the protein ex- pression of EMMPRIN in the pouches was detected by using the reverse - transcription polymerase chain reaction and immunohis- tochemistry methods. The NF - KBP65expression in the pouches was detected by using western blotting. [ Results] A white and crisp subcutaneous pouch was noted on the back. Extensive cell infiltration was detected by using a light microscope. However, only a piece of limpid subcutaneous pouch was noted on the back of the mice in group A. The EMMPRIN gene and protein ex- pression reached its peak on day 14, with the expression of EMMPRIN and NF - KBP65in group C being lesser than in group B （P 〈 0. 05） and greater than in group A （P 〈 0. 01 ） . [ Conclusion ] EMMPRIN expression is induced by particulates and changes with time. EMMPRIN generation may be regulated by NF - KBP 65.