中国全科医学2016，Vol.19Issue(33)：4079-4082,4.DOI:10.3969/j.issn.1007-9572.2016.33.011
经尿道灌注干细胞白血病基因慢病毒转染豚鼠糖尿病膀胱病变的效果研究
Effect of SCL Lentivirus Transfection Through Perfusing Diabetic Cystopathy of Bladder in Guinea Pig Models
摘要
Abstract
Objective To prepare guinea pig model of diabetic cystopathy( DCP)of bladder and transurethrally perfuse stem cell leukemia(SCL)gene lentivirus in order to evaluate the transfection efficiency and stability. Methods From November 2014 to June 2015,a total of 60 guinea pigs were selected from 75 Holland male guinea pigs by random number table method and given streptozotocin(STZ)(200 mg/ kg)by a single intraperitoneal injection. The models of diabetic were screened according to the blood glucose level > 11. 1 mmol/ L after 6 - week feeding. Then the models of DCP were screened by urodynamic testing after 4 - week feeding. The left 15 guinea pigs were injected with the same dose of buffer STZ and fed in the same way(the control group). Fifteen guinea pigs were selected from the successful DCP models and divided into Ⅰ group( n = 3)and experimental group(n = 12)by random number table method. The Ⅰgroup was not treated,and the experimental group was transurethrally perfused with SCL lentivirus(1. 6 × 107 TU)twice. At the 2th,7th,14th,28th day after the second perfuse,3 guinea pigs were killed by cervical dislocation and marked as Ⅱ,Ⅲ,Ⅳ and Ⅴ group,respectively. The Ⅰ group was killed at the same time with Ⅱ group. The bladder tissue was frozen in liquid nitrogen. The distribution of green fluorescence in the tissue was observed under laser confocal microscopy after glycerol sealing. The expression level of SCL mRNA was detected using qRT - PCR method. Results During the preparing the DCP model,18 guinea pigs were discarded in the diabetic model screening and 19 guinea pigs were discarded in the urodynamic testing,and finally 23 DCP models were obtained(DCP model group). The maximal detrusor pressure in DCP model group of guinea pigs was lower than control group,the maximum bladder capacity was larger than control group(P < 0. 01). There was no green fluorescence in the bladder tissue layer of Ⅰ group. The green fluorescence was not obvious in the bladder tissue layer of Ⅱ group. A super strong green fluorescence was observed throughout the whole bladder tissue layer in Ⅲgroup. There was strong green fluorescence in the whole bladder tissue layer of Ⅳgroup. There was still visible green fluorescence in the bladder tissue layer of Ⅴgroup. The expression of SCL mRNA in Ⅰ group was(1. 00 ± 0. 00). The expression of SCL mRNA in Ⅱgroup was(1. 22 ± 0. 06)times higher than Ⅰ group. The expression of SCL mRNA in Ⅲ group was(50. 45 ± 3. 17)times higher than Ⅰ group. The expression of SCL mRNA in Ⅳgroup was(24. 75 ± 6. 49)times higher than Ⅰ group. The expression of SCL mRNA in Ⅴ group was(3. 27 ± 1. 00)times higher than Ⅰgroup. Conclusion The lentiviral vector of SCL gene could successfully transfect into the DCP bladder of guinea pig and stably express,which might bring a new way for the clinical treatment of DCP.关键词
糖尿病并发症/膀胱疾病/慢病毒感染/干细胞白血病Key words
Diabetes complications/Urinary bladder diseases/Lentivirus infections/Stem cell leukemia分类
医药卫生引用本文复制引用
魏艳青,马路平,王江平,钱彪,王勤章..经尿道灌注干细胞白血病基因慢病毒转染豚鼠糖尿病膀胱病变的效果研究[J].中国全科医学,2016,19(33):4079-4082,4.基金项目
国家自然科学基金资助项目 （）
