Local delivery of matrix metalloproteinase gene prevents the onset of renal sclerosis in streptozotocin-induced diabetic mice
Local delivery of matrix metalloproteinase gene prevents the onset of renal sclerosis in streptozotocin-induced diabetic mice
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DOI：
10.1089/10763270360728206
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发表时间：
2003-12-01
影响因子：
--
通讯作者：
Tabata, Y
中科院分区：
文献类型：
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作者：
Aoyama, T;Yamamoto, S;Tabata, Y
本研究旨在调查基质金属蛋白酶（MMP）是否具有预防进行性肾病中破坏性纤维化发生的功能。作为质粒DNA的缓释载体，通过胺化制备的阳离子明胶配制可生物降解的水凝胶和微球。由于水凝胶降解，质粒 DNA 从阳离子明胶水凝胶中释放出来。构建了包含巨细胞病毒启动子和人重组MMP-1基因(pCMV-MMP)的质粒DNA。将掺有pCMV-MMP的明胶微球以及含或不含pCMV-MMP的磷酸盐缓冲盐水（PBS）注射到C57BL/6小鼠的肾下囊中，术后7天腹腔注射链脲佐菌素（STZ）诱导糖尿病。注射 STZ 后 4 周处死小鼠，采集血液和肾脏样本进行生化和组织学检查。免疫荧光研究证实，在注射了掺有 pCMV-MMP 的明胶微球的肾组织周围表达了 MMP 蛋白。当使用掺有pCMV-MMP的阳离子明胶微球时，小鼠的血液尿素氮水平显着低于其他组。组织学观察到给予掺有 pCMV-MMP 的明胶微球的小鼠肾脏中胶原蛋白含量降低。此外，羟脯氨酸测定显示肾脏中羟脯氨酸的含量显着降低。我们得出的结论是，持续释放 MMP-1 基因对于 STZ 诱导的糖尿病小鼠模型中的肾纤维化和功能障碍是一项有前景的预防性试验。
The present study was undertaken to investigate whether matrix metalloproteinase (MMP) functions to prevent the occurrence of destructive fibrosis in progressive renal disease. As a sustained release carrier of plasmid DNA, biodegradable hydrogels and microspheres were formulated from cationized gelatin prepared through aminization. Plasmid DNA was released from the cationized gelatin hydrogels as a result of hydrogel degradation. A plasmid DNA including a cytomegalovirus promoter and human recombinant MMP-1 gene (pCMV-MMP) was constructed. Gelatin microspheres incorporating pCMV-MMP as well as phosphate-buffered saline (PBS) with or without pCMV-MMP were injected into the renal subcapsule of C57BL/6 mice, which were intraperitoneally injected with streptozotocin (STZ) to induce diabetes 7 days after operation. The mice were killed 4 weeks after STZ injection to sample their blood and kidneys for biochemical and histological examinations. An immunofluorescence study confirmed that MMP protein was expressed around the renal tissue injected with gelatin microspheres incorporating pCMV-MMP. When applied with cationized gelatin microspheres incorporating pCMV-MMP, the mice showed a level of blood urea nitrogen significantly lower than that of other groups. A reduced content of collagen in the kidneys of mice administered gelatin microspheres incorporating pCMV-MMP was histologically observed. Further, the hydroxyproline assay revealed a significantly decreased content of hydroxyproline in kidney. We conclude that sustained release of MMP-1 gene is a promising prophylactic trial for kidney fibrolysis and dysfunction in the STZ-induced diabetic mouse model.