Experimental study for gene induction therapy to temporomandibular arthritis

我们从U937细胞中构建了人白介素1受体拮抗剂(hIL-1ra)基因的表达载体，并在体外和体内实验中证实了其表达。利用体内基因电穿孔技术将构建的基因导入正常膝关节和颞下颌关节。Western blotting证实这些关节中有hIL-1ra的表达，但表达太弱，不能减轻关节炎的病情。因此，我们试图将该基因导入包括关节周围肌肉组织在内的关节周围组织中，以增强大鼠佐剂性关节炎的抗炎蛋白的产生。肌肉电穿孔基因诱导和hIL-1ra的表达：将不同浓度的DNA溶液(包括构建的载体)注入膝关节周围的股骨肌肉。HIL-1ra的表达以50微克注射组最高。基因表达持续时间：电穿孔基因诱导后1~28天(50微克组)可观察到hIL-1ra的表达。结论：从我们的研究来看，将IL-1ra基因诱导到关节周围组织将是治疗关节炎的一种可能的选择，因为关节周围肌肉具有产生抗炎蛋白的生物反应器效应。

We constructed a plasmid vector of human interleukin 1 receptor antagonist (hIL-1ra) gene from U937 cell and confirmed the expression in in-vitro and in-vivo studies. The constructed gene was used for gene induction into the normal knee and the temporomandibular joints using with in-vivo gene electroporation technique. The production of hIL-1ra from those joints was confirmed with Western blotting but the production was too weak to reduce the arthritic condition.Therefore we tried to introduce the gene into peri-joint tissues including muscle tissue around the joint to enhance the anti-inflammatory protein production in adjuvant arthritis of the rat.1. Electroporatic gene induction and expression of hIL-1ra to a muscle : Various injected concentrations of DNA solution including the constructed vector were tested into the femoral muscle around the knee joint. The highest expression of the hIL-1ra was observed in the 50 micrograms' plasmid injection group.2. Duration of the gene expression : The expression of hIL-1ra was observed from one day to 28 days after electroporatic gene induction (50 micrograms' group).Conclusion : From our studies, electroporatic IL-1ra gene induction to the peri-joint tissues will be a possible option of the management of arthritic conditions because peri-joint muscle has a bioreactor effect to produce anti-inflammatory protein.