Shen Maolei, Qian Biao, Xu Hao, Fu Jiahao, Wang Yuchen, Wang Qinzhang
Department of Urology, First Affiliated Hospital, Medical College, Shihezi University, Shihezi Xinjiang 832000, China.
Health Center, PLA 69235, Kuitun Xinjiang 833200, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2019 Feb 28;44(2):117-121. doi: 10.11817/j.issn.1672-7347.2019.02.001.
To determine the effect of a recombinant lentivirus containing human stem cell leukemia (SCL) gene on the expression of c-kit protein in damaged interstitial cells of Cajal (ICC) under high glucose condition. Methods: After isolation of ICC, the cells were cultured for 24 hours until the cells were adherent. After identification by inverted microscope and immunofluorescence, ICC cells were divided into two groups: A control group and a high glucose group. The control group was added with a medium containing 5 mmol/L of glucose. The high glucose group was added with a medium containing 20 mmol/L of glucose. After 48 h of continuous cultivation, the high glucose group was divided into 3 subgroups: A blank group, an empty lentivirus group, and an experimental group. The blank group, the empty lentivirus group, and the experimental group were added a medium containing PBS solution, empty lentivirus, and a recombinant lentivirus containing the SCL gene with a glucose concentration of 5 mmol/L, respectively. The cultures were incubated for 24 and 48 h. The expression of c-kit protein in ICC in each group was detected by Western blot. Results: After 24 or 48 h, the expression of c-kit protein in ICC was significantly lower in the blank group and the lentivirus group than that in the control group, and the expression of c-kit protein in ICC was significantly higher in the experimental group than that in the blank group and the empty lentivirus group, but it was still lower than that in the control group (all P<0.05). Conclusion: The recombinant lentivirus of SCL gene can up-regulate the expression of c-kit protein in functionally impaired ICC under high glucose condition.
探讨含人干细胞白血病（SCL）基因的重组慢病毒对高糖条件下受损的 Cajal 间质细胞（ICC）中 c-kit 蛋白表达的影响。方法：分离 ICC 细胞后，培养 24 小时至细胞贴壁。经倒置显微镜和免疫荧光鉴定后，将 ICC 细胞分为两组：对照组和高糖组。对照组加入含 5 mmol/L 葡萄糖的培养基，高糖组加入含 20 mmol/L 葡萄糖的培养基。连续培养 48 小时后，将高糖组分为 3 个亚组：空白组、空慢病毒组和实验组。空白组、空慢病毒组和实验组分别加入含 PBS 溶液、空慢病毒和含 SCL 基因的重组慢病毒且葡萄糖浓度为 5 mmol/L 的培养基。培养 24 和 48 小时。采用 Western blot 检测各组 ICC 中 c-kit 蛋白的表达。结果：培养 24 或 48 小时后，空白组和慢病毒组 ICC 中 c-kit 蛋白的表达显著低于对照组，实验组 ICC 中 c-kit 蛋白的表达显著高于空白组和空慢病毒组，但仍低于对照组（均 P<0.05）。结论：SCL 基因重组慢病毒可上调高糖条件下功能受损的 ICC 中 c-kit 蛋白的表达。
