颞下颌关节腔内基因转染的生物安全性实验研究

A Study on Biological Security of Gene Transfection in the Temporomandibular Joint Cavity

目的研究阳离子脂质体介导人白介素-1受体拮抗剂(hIL-1Ra)基因在颞下颌关节腔内基因转染后对全身其它脏器的影响。方法20只新西兰白兔随机分为4组,每组5只。第一、二、三组颞下颌关节腔内连续3次间隔24h注射生理盐水,第一组的生理盐水内含50μL阳离子脂质体介导的100μgpcDNA3.1-hIL-1Ra基因的质粒,第二组的生理盐水内含100μL阳离子脂质体介导的200μgpcDNA3.1-hIL-1Ra基因的质粒,第三组的生理盐水内含50μL阳离子脂质体介导的100μgpcDNA3.1的质粒,第四组不进行任何处理,注射完4周后处死所有动物。各组兔获取颞下颌关节、心、肝、脾、肺、肾、胰标本。首先行大体观察有无变化,接着取部分关节和脏器组织用逆转录-聚合酶链反应法(RT-PCR)来检测是否有hIL-1Ra基因的表达,最后行组织学观察。结果各组大体观未见异常,经RT-PCR法证实第一、二组颞下颌关节组织有hIL-1Ra基因的表达,而第三、四组颞下颌关节组织及各组其它脏器均无hIL-1Ra基因的表达,各组脏器标本无明显组织病理变化。结论阳离子脂质体介导hIL-1Ra基因进行兔颞下颌关节腔内基因转染,有良好的生物安全性。

Objective To study the possible effect of cationic lipid mediated human Interleukin 1 Receptor Antagonist gene transfection on the other organs through the direct delivery in the temporomandibular joint cavity. Methods Twenty Newzerland rabbits were divided into four groups. Group 1 and 2 received three consecutive injections at 24-hour intervals of 0.9% saline containing 100 and 200 μg of hIL-1Ra plasmid and lipid. Group 3 received three consecutive intra-articular injections at 24-hour intervals of 0.9% saline containing a cationic lipid, and a DNA plasmid. Group 4 received no treatment. All rabbits were sacrificed after 4 weeks. TMJ, liver, kidney, heart, spleen, lung, and pancreas were harvested. RT-PCR was applied to examine the expression of hIL-1Ra gene in these organs. Macroscopy and histologic evalution were also performed. Results The expression of hIL-1Ra mRNA in TMJ of experimental group 1 and 2 were detected by RT-PCR but not in any other organs. Pathologic disorder was found neither by macroscopic observation nor histological examination. Conclusion Intra-articular direct injection can achieve gene delivery limited to the joint cavity, possibly eliminating unnecessary systemic effects.