Observation on number, distribution and ultrastructure of interstitial cells of Cajal in guinea pig with DCP after transurethral perfusion of SCL recombinant lentiviral vector

WANG Zixiong; WANG Qinzhang; QIAN Biao

doi:10.3969/j.issn.1002-266X.2017.15.007

Abstract：Objective To observe morphological changes in the number, distribution and ultrastructure of interstitial cells of Cajal (ICC) in guinea pig with diabetic cystopathy (DCP) after transurethral perfusion of stem cell leukemia (SCL) gene recombinant lentiviral vector. Methods We constructed the recombinant lentiviral vector of SCL gene and demarcated its titer. Seventy guinea pigs were selected and a single intraperitoneal injection of streptozotocin (STZ) was used to construct the DCP models of guinea pigs at the end of the 12th week. Twenty-seven successful DCP models of guinea pigs were randomly divided into the 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 injected with SCL gene recombinant lentivirus, empty lentivirus and the same amount of PBS solution, respectively. On day 7, 14 and 28 after transfection, 3 guinea pigs were sacrificed from each group to obtain the bladder tissue to make the section. The expression of GFP, bladder ICC number and distribution was observed under laser confocal microscope, and the ultrastructural changes of bladder ICC were observed by transmission electron microscope. Results In the experimental group, the number of ICC increased with the increase of transfection time (P<0.01), and the distribution was more and more intensive, but the green fluorescence decreased gradually. The blank group and the control group had no green fluorescence, and the number of ICC decreased with the increase of transfection time (P<0.05) and the distribution was sparse; on day 14 and 28, the number of ICC in the experimental group was higher than that in the blank group and the control group (all P<0.01). In the experimental group, with the transfection time prolonged, the number of ICC cells increased, morphology tended to normal, intracellular vacuole decreased, and the ultrastructure of ICC continuously recovered; however, in the positive control group and blank control group, ICC pathological ultrastructure changes gradually aggravated. Conclusions After the SCL gene is transfected into the bladder of DCP guinea pig, the number of ICC increases, the distribution becomes more intensive, and the ultrastructure of the cells is repaired. It suggests that SCL gene might have a certain therapeutic effect on DCP.