SCL基因重组慢病毒上调高糖环境下Cajal间质细胞中c-kit蛋白表达

目的：探讨含人干细胞白血病(stem cell leukemia，SCL)基因重组慢病毒对高糖环境下受损Cajal间质细胞(interstitial cells of Cajal，ICC)中酪氨酸激酶受体(c-kit)蛋白表达的影响。方法：分离ICC，培养24 h贴壁，经倒置显微镜、免疫荧光鉴定后，将ICC细胞分为对照组、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，培养48 h后；对照组不变，高糖组分成空白组、空慢病毒组和实验组3个亚组，3组分别加入PBS、空慢病毒和含SCL基因重组慢病毒5 mmol/L葡萄糖的培养基，继续培养24和48 h，采用Western印迹法检测两个时段各组中ICC中c-kit 蛋白表达情况。结果：24或48 h后，空白组、空慢病毒组中ICC的c-kit蛋白表达水平均明显低于对照组，实验组中ICC中c-kit蛋白的表达水平明显均高于空白组和空慢病毒组，但是仍低于对照组(均P<0.05)。结论：SCL基因重组慢病毒可以上调高糖环境下功能受损ICC中c-kit蛋白表达量。

Effect of recombinant lentivirus of SCL gene on expression of c-kit protein in interstitial cells of Cajal under high glucose condition

Objective: To determine the effect of a recombinant lentivirus containing human stem cellleukemia (SCL) gene on the expression of c-kit protein in damaged interstitial cells of Cajal (ICC)under high glucose condition.Methods: Aft er isolation of ICC, the cells were cultured for 24 hours until the cells were adherent.Aft er identifi cation by inverted microscope and immunofl uorescence, ICC cells were divided intotwo groups: A control group and a high glucose group. The control group was added with a mediumcontaining 5 mmol/L of glucose. The high glucose group was added with a medium containing20 mmol/L of glucose. After 48 h of continuous cultivation, the high glucose group was dividedinto 3 subgroups: A blank group, an empty lentivirus group, and an experimental group. The blankgroup, the empty lentivirus group, and the experimental group were added a medium containingPBS solution, empty lentivirus, and a recombinant lentivirus containing the SCL gene with aglucose concentration of 5 mmol/L, respectively. The cultures were incubated for 24 and 48 h. Theexpression 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 blankgroup and the lentivirus group than that in the control group, and the expression of c-kit protein inICC was significantly higher in the experimental group than that in the blank group and the emptylentivirus 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-kitprotein in functionally impaired ICC under high glucose condition.