PDF(Sci-hub)
Abstract：
Diabetic cystopathy (DCP) is a chronic complication of diabetes mainly within the submucosal and muscular layers of the bladder due to the hyperglycemia-induced ischemia. As no effective therapies are currently available, the administration of optimized mesenchymal stem cells (MSCs) provides a potential treatment of DCP. Thus far, new strategy, such as genetic modification of MSCs, has been developed and has shown promising outcomes of various disorders.
This study was conducted using integrin-linked kinase (ILK) gene-modified bone marrow-derived stem cells (BMSCs) for streptozotocin (STZ)-induced diabetic cystopathy in a rat model. In total, 68 male Sprague-Dawley rats were randomized into five groups: sham control (control group, n = 10); DCP model alone (DM group, n = 10); DCP rats intravenously treated with BMSCs (BMSC group, n = 16); DCP rats accepted adenoviral vector-infected BMSCs (Ad-null-BMSC group, n = 16) and DCP rats accepted ILK adenoviral vector-infected BMSCs (Ad-ILK-BMSC group, n = 16). Diabetic rats accepted cell transplantation in the experimental group (2 rats per group) were sacrificed for the bladder tissue on the third day, 7th day, and 14th day of treatment respectively ahead of schedule. At 4 weeks after treatment, all rats in five groups accepted urodynamic studies to evaluate bladder function and were sacrificed for bladder tissue.
Our data showed that the underactive bladder function was significantly improved in DCP rats intravenously treated with ILK gene-modified BMSCs compared to those in the DM, BMSCs, and Ad-null-BMSC group. Meanwhile, we found that gene-modified BMSC treatment significantly promoted the activation of the AKT/GSK-3β pathway by increasing phosphorylation and led to the enhancement of survival. In addition, the expression levels of angiogenesis-related protein vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and stromal cell-derived factor-1 (SDF-1) were significantly higher in the Ad-ILK-BMSC group than that in the DM, BMSCs, and Ad-null-BMSC group as assessed by enzyme-linked immunosorbent assay and western blot. As two indicators of vascular endothelial cell markers, the expression of von Willebrand factor (vWF) and CD31 by western blot and immunofluorescent staining revealed that the percentage of the vascular area of the bladder tissue significantly increased in Ad-ILK-BMSC group compared with the BMSCs and Ad-null-BMSC group on the 14th day of treatment. Histological and immunohistochemical staining (hematoxylin and eosin (HE), vWF, Ki67, and TUNNEL) on the bladder tissue revealed statistically different results between groups.
ILK gene-modified BMSCs restored the bladder function and histological construction via promoting the process of angiogenesis and protecting cells from high glucose-associated apoptosis in STZ-induced DCP rat model, which provides a potential for the treatment of patients with DCP.
This study was conducted using integrin-linked kinase (ILK) gene-modified bone marrow-derived stem cells (BMSCs) for streptozotocin (STZ)-induced diabetic cystopathy in a rat model. In total, 68 male Sprague-Dawley rats were randomized into five groups: sham control (control group, n = 10); DCP model alone (DM group, n = 10); DCP rats intravenously treated with BMSCs (BMSC group, n = 16); DCP rats accepted adenoviral vector-infected BMSCs (Ad-null-BMSC group, n = 16) and DCP rats accepted ILK adenoviral vector-infected BMSCs (Ad-ILK-BMSC group, n = 16). Diabetic rats accepted cell transplantation in the experimental group (2 rats per group) were sacrificed for the bladder tissue on the third day, 7th day, and 14th day of treatment respectively ahead of schedule. At 4 weeks after treatment, all rats in five groups accepted urodynamic studies to evaluate bladder function and were sacrificed for bladder tissue.
Our data showed that the underactive bladder function was significantly improved in DCP rats intravenously treated with ILK gene-modified BMSCs compared to those in the DM, BMSCs, and Ad-null-BMSC group. Meanwhile, we found that gene-modified BMSC treatment significantly promoted the activation of the AKT/GSK-3β pathway by increasing phosphorylation and led to the enhancement of survival. In addition, the expression levels of angiogenesis-related protein vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and stromal cell-derived factor-1 (SDF-1) were significantly higher in the Ad-ILK-BMSC group than that in the DM, BMSCs, and Ad-null-BMSC group as assessed by enzyme-linked immunosorbent assay and western blot. As two indicators of vascular endothelial cell markers, the expression of von Willebrand factor (vWF) and CD31 by western blot and immunofluorescent staining revealed that the percentage of the vascular area of the bladder tissue significantly increased in Ad-ILK-BMSC group compared with the BMSCs and Ad-null-BMSC group on the 14th day of treatment. Histological and immunohistochemical staining (hematoxylin and eosin (HE), vWF, Ki67, and TUNNEL) on the bladder tissue revealed statistically different results between groups.
ILK gene-modified BMSCs restored the bladder function and histological construction via promoting the process of angiogenesis and protecting cells from high glucose-associated apoptosis in STZ-induced DCP rat model, which provides a potential for the treatment of patients with DCP.
摘要：
糖尿病性膀胱病（DCP）是糖尿病的慢性并发症，主要是由于高血糖引起的缺血而在膀胱的粘膜下层和肌肉层内。由于目前没有有效的治疗方法，优化的间充质干细胞（MSC）的给药提供了DCP的潜在治疗。到目前为止,新战略,如MSCs的遗传修饰，已被开发，并已显示出各种疾病的有希望的结果。
本研究是使用整合素连接激酶（ILK）基因修饰的骨髓来源干细胞（BMSCs）在大鼠模型中链脲佐菌素（STZ）诱导的糖尿病性膀胱病变进行的。总的来说,68只雄性SD大鼠随机分为5组：假对照组（对照组，n=10)；单独DCP模型(DM组，n=10）；DCP大鼠静脉注射BMSCs（BMSC组，n=16）；DCP大鼠接受腺病毒载体感染的BMSCs（Ad-null-BMSC组，n=16）和DCP大鼠接受ILK腺病毒载体感染的BMSCs（Ad-ILK-BMSC组，n=16)。实验组接受细胞移植的糖尿病大鼠(每组2只)于第3天处死膀胱组织,第七天,和第14天的治疗分别提前。治疗后4周,五组大鼠均接受尿流动力学检查以评估膀胱功能,并处死膀胱组织.
我们的数据表明，与DM相比，用ILK基因修饰的BMSCs静脉内治疗的DCP大鼠的膀胱功能不足明显改善，BMSCs,和Ad-null-BMSC组。同时,我们发现基因修饰的BMSC处理通过增加磷酸化显著促进AKT/GSK-3β通路的激活,并导致存活率的提高.此外,血管生成相关蛋白血管内皮生长因子(VEGF)的表达水平,碱性成纤维细胞生长因子（bFGF），Ad-ILK-BMSC组的基质细胞衍生因子-1（SDF-1）明显高于DM组，BMSCs,和Ad-null-BMSC组，通过酶联免疫吸附测定和蛋白质印迹评估。作为血管内皮细胞标志物的两项指标,Westernblot和免疫荧光染色显示，与BMSCs和Ad-null-BMSC组相比，Ad-ILK-BMSC组膀胱组织血管面积百分比显着增加。治疗第14天。组织学和免疫组织化学染色（苏木精和伊红（HE），vWF,膀胱组织上的Ki67和TUNNEL）显示两组之间具有统计学差异的结果。
ILK基因修饰的BMSCs通过促进血管生成过程和保护细胞免受高糖诱导的DCP大鼠模型的凋亡，从而恢复膀胱功能和组织学结构。这为DCP患者的治疗提供了潜力。
本研究是使用整合素连接激酶（ILK）基因修饰的骨髓来源干细胞（BMSCs）在大鼠模型中链脲佐菌素（STZ）诱导的糖尿病性膀胱病变进行的。总的来说,68只雄性SD大鼠随机分为5组：假对照组（对照组，n=10)；单独DCP模型(DM组，n=10）；DCP大鼠静脉注射BMSCs（BMSC组，n=16）；DCP大鼠接受腺病毒载体感染的BMSCs（Ad-null-BMSC组，n=16）和DCP大鼠接受ILK腺病毒载体感染的BMSCs（Ad-ILK-BMSC组，n=16)。实验组接受细胞移植的糖尿病大鼠(每组2只)于第3天处死膀胱组织,第七天,和第14天的治疗分别提前。治疗后4周,五组大鼠均接受尿流动力学检查以评估膀胱功能,并处死膀胱组织.
我们的数据表明，与DM相比，用ILK基因修饰的BMSCs静脉内治疗的DCP大鼠的膀胱功能不足明显改善，BMSCs,和Ad-null-BMSC组。同时,我们发现基因修饰的BMSC处理通过增加磷酸化显著促进AKT/GSK-3β通路的激活,并导致存活率的提高.此外,血管生成相关蛋白血管内皮生长因子(VEGF)的表达水平,碱性成纤维细胞生长因子（bFGF），Ad-ILK-BMSC组的基质细胞衍生因子-1（SDF-1）明显高于DM组，BMSCs,和Ad-null-BMSC组，通过酶联免疫吸附测定和蛋白质印迹评估。作为血管内皮细胞标志物的两项指标,Westernblot和免疫荧光染色显示，与BMSCs和Ad-null-BMSC组相比，Ad-ILK-BMSC组膀胱组织血管面积百分比显着增加。治疗第14天。组织学和免疫组织化学染色（苏木精和伊红（HE），vWF,膀胱组织上的Ki67和TUNNEL）显示两组之间具有统计学差异的结果。
ILK基因修饰的BMSCs通过促进血管生成过程和保护细胞免受高糖诱导的DCP大鼠模型的凋亡，从而恢复膀胱功能和组织学结构。这为DCP患者的治疗提供了潜力。
