用重组表达体系pCKM-mPTH质粒治疗甲状旁腺功能低下症的实验研究

pCKM-mPTH recombinant plasmid therapy for hypoparathyroidism: an in vivo experimental research with rats

目的研究重组pCKMmPTH质粒在SD大鼠甲状旁腺功能低下症模型中的治疗作用。方法将30只SD大鼠随机分为3组,A组为正常组,肌肉注射生理盐水,B组、C组制作甲状旁腺功能低下症模型后,分别肌肉注射pcDNA31(+)空质粒和pCKMmPTH质粒。用放免法测定不同时间的血清甲状旁腺激素(PTH)浓度。结果动物模型在注射pCKMmPTH质粒后,其血清PTH浓度在近1个月内显著高于注射前及空质粒对照组,未观察到有甲状旁腺功能低下症的表现。结论应用pCKMmPTH质粒治疗SD大鼠的甲状旁腺功能低下症是有效的,对未来的临床应用研究有一定的指导意义。

Objective To investigate the effect of gene therapy with pCKM mPTH recombinant plasmid on hypoparathyroidism Methods Thirty male Sprague Dawley rats were randomly divided into three groups: group A, undergoing fake operation and then injection of normal saline into the quadriceps muscle of the left leg; group B, undergoing parathyroidectomy and injection of blank pcDNA3 1(+) plasmid into the quadriceps muscle of the left leg; and group C, undergoing parathyroidectomy injection of pCKM mPTH plasmid containing gene of mutated proparat hormone into the quadriceps muscle of the left leg Peripheral blood was collected from each rat before operation and 3, 5, 6, 7, 9, 12, 15, 20 and 30 days after operation Radioimmunoassay was used to detect the level of parathyroid hormone Results The level of paratheproid hormone (PTH) of the group B was 10 61 ng/dl±0 86 ng/dl before injection of blank plasmid and gradually decreased after the injection, for example, 10 39 ng/dl±0 36 ng/dl 1 day after the injection and 9 45 ng/dl±2 64 ng/dl 60 days after the injection The level of PTH of the group C was 10 75 ng/dl± 0 45 ng/dl before the injection of pCKM mPTH plasmid and gradually increased after the injection, for example, 28 39±0 22 ng/dL 1 day after the injection, significantly higher than that before the injection, peaked on the 6th day (43 28 ng/dl±2 45 ng/dl), then gradually decreased, and reached 15 32 ng/dl±0 26 ng/dl on the 60th day However, the levels of PTH in the group C at any time point were all significantly higher than those of the group B (all P <0 01) Symptoms of hypoparathyroidism were not observed in the rats of the group C Conclusion pCKM mPTH recombinant plasmid therapy is effective in treatment of hypoparathyroidism