经尿道灌注SCL基因重组慢病毒的DCP豚鼠膀胱Cajal样间质细胞数量、分布及超微结构观察

发布时间：2018-04-19 03:00

本文选题：干细胞白血病基因 + Cajal样间质细胞　；参考：《山东医药》2017年15期

【摘要】：目的观察经尿道灌注干细胞白血病(SCL)基因重组慢病毒的糖尿病膀胱病(DCP)豚鼠膀胱Cajal样间质细胞(ICC)的数量、分布及超微结构变化。方法构建SCL基因重组慢病毒且标定滴度。选择雄性豚鼠70只,单次腹腔注射链尿佐菌素12周后构建DCP豚鼠模型,将成功造模的27只豚鼠随机分为实验组、阳性对照组、空白对照组,每组9只。实验组、阳性对照组、空白对照组豚鼠分别经尿道灌注(转染)SCL基因重组慢病毒、空慢病毒和等量PBS液。分别于转染后7、14、28 d每组各处死3只豚鼠,取膀胱组织,制作切片,激光共聚焦显微镜下观察GFP表达、膀胱ICC数量及分布变化,透射电镜下观察膀胱ICC的超微结构变化。结果实验组随转染时间延长,ICC数量逐渐增多(P均0.01),分布越发密集,但绿色荧光逐渐减退;空白组和对照组未见绿色荧光,随转染时间延长,ICC数量逐渐减少(P均0.05),分布稀疏;转染14、28 d时,实验组ICC数量较空白组和对照组增加(P均0.01)。实验组随转染时间延长,ICC细胞器数量增多,形态趋于正常,细胞内空泡减少,胞周突起延长变多,ICC超微结构处于不断恢复之中;而阳性对照组和空白对照组ICC超微结构病理变化逐渐加重。结论经尿道灌注SCL基因重组慢病毒转染DCP豚鼠膀胱后,膀胱ICC数量增多,分布更加密集,细胞超微结构得到修复,提示SCL基因对DCP可能有一定治疗效果。
[Abstract]:Objective to observe the number, distribution and ultrastructure of Cajal like interstitial cells (ICCs) in bladder of guinea pigs perfused with STL gene recombinant lentivirus by transurethral instillation of stem cell leukemia (SCL) gene in guinea pig bladder.Methods Recombinant lentivirus of SCL gene was constructed and its titer was calibrated.DCP guinea pig model was established by single intraperitoneal injection of streptozotocin for 12 weeks in 70 male guinea pigs. 27 guinea pigs were randomly divided into three groups: experimental group, positive control group and blank control group (n = 9 in each group).The experimental group, the positive control group and the blank control group were perfused by transurethral perfusion (transfection) of recombinant lentivirus, empty lentivirus and the same amount of PBS solution.Three guinea pigs were killed in each group on the 28th day after transfection. The bladder tissue was taken and sections were made. The expression of GFP, the number and distribution of ICC in bladder were observed under confocal laser microscope, and the ultrastructural changes of ICC in bladder were observed under transmission electron microscope.Results with the extension of transfection time, the number of ICC in the experimental group increased gradually (P < 0.01) and the distribution became denser, but the green fluorescence decreased gradually, but no green fluorescence was found in the blank group and the control group, and the number of ICC decreased gradually with the extension of the transfection time, and the distribution was sparse.On the 28th day after transfection, the number of ICC in the experimental group was higher than that in the blank group and the control group (P < 0.01).In the experimental group, the number of ICC organelles increased with the extension of transfection time, the morphology tended to be normal, the vacuoles in the cells decreased, and the ultrastructure of ICC was in the process of recovery.The ultrastructural pathological changes of ICC in the positive control group and the blank control group were gradually aggravated.Conclusion after transurethral infusion of SCL gene recombinant lentivirus into the bladder of DCP guinea pigs, the number and distribution of ICC in the bladder increased, and the ultrastructure of the cells was repaired, suggesting that SCL gene may have a therapeutic effect on DCP.
【作者单位】：石河子大学医学院附属第一医院;
【基金】：国家自然科学基金资助项目(81360120)
【分类号】：R587.2;R694

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