Department of Neurosurgery, Dongguan People's Hospital, Affiliated Dongguan People's Hospital of Southern Medical University, Dongguan, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(8):3198-3205. doi: 10.26355/eurrev_201904_17678.
To explore the inhibitory effect of transforming growth factor-beta (TGF-β) gene modified human amniotic mesenchymal stem cells on rejection after xenotransplantation of peripheral nerves.
In this study, 6 placentas collected in our hospital were selected as the source of human amniotic mesenchymal stem cells. A total of 60 C57BL/6 experimental mice (mouse sciatic nerves were removed before the experiment) were taken as research objects. Mice were randomly divided into experimental group 1, experimental group 2 and experimental group 3 (xenogenous peripheral nerves were introduced to all experimental groups), and a control group (autologous peripheral nerves were introduced). Among them, TGF-β gene modified (overexpression) human amniotic mesenchymal stem cells were introduced to experimental group 1; TGF-β gene modified (inhibition) human amniotic mesenchymal stem cells were introduced to experimental group 2; normal human amniotic mesenchymal stem cells were introduced to experimental group 3; and autologous sciatic nerves were introduced to control group. The messenger ribonucleic acid (mRNA) and protein expressions of the TGF-β in different human amniotic mesenchymal stem cells were detected by quantitative polymerase chain reaction (qPCR) and Western blotting, respectively. Mouse sciatic nerve function in each group after 2 weeks of procedures was detected via the CatWalk system. Expression level of interleukin-22 (IL-22) in the peripheral tissues of transplanted nerves and blood was detected using immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). Its mRNA level was examined via fluorescence quantitative PCR.
TGF-β1 was highly expressed in mice of experimental group 1, but lowly expressed in experimental group 2 relative to that of experimental group 3 (p<0.05). CatWalk test results revealed that the main indexes in experimental group 1 were superior to those in other groups, while the main indexes in experimental group 2 were inferior to those in other groups. According to immunohistochemistry and ELISA results, there were significant differences in the expression level of IL-22 in mice of different treatment groups (p<0.05). IL-22 level was the lowest in control group [(5.05±0.15) pg/mL], followed by that in experimental group 1 [(6.52±0.24) pg/mL], and it was the highest in experimental group 2 [(9.47±0.31) pg/mL].
Human amniotic mesenchymal stem cells overexpressing TGF-β can inhibit rejection after xenotransplantation of peripheral nerves.
探讨转化生长因子-β（TGF-β）基因修饰的人羊膜间充质干细胞对异种周围神经移植排斥反应的抑制作用。
本研究选择我院采集的 6 例胎盘作为人羊膜间充质干细胞的来源，取 60 只 C57BL/6 实验小鼠（实验前切除小鼠坐骨神经）作为研究对象。将小鼠随机分为实验组 1、实验组 2 和实验组 3（所有实验组均引入异种周围神经）和对照组（引入自体周围神经）。其中实验组 1 导入 TGF-β 基因修饰（过表达）的人羊膜间充质干细胞；实验组 2 导入 TGF-β 基因修饰（抑制）的人羊膜间充质干细胞；实验组 3 导入正常的人羊膜间充质干细胞；对照组导入自体坐骨神经。通过实时定量聚合酶链反应（qPCR）和 Western blot 分别检测不同人羊膜间充质干细胞中 TGF-β 的信使核糖核酸（mRNA）和蛋白表达。术后 2 周，采用 CatWalk 系统检测各组小鼠的坐骨神经功能。采用免疫组织化学法和酶联免疫吸附试验（ELISA）检测移植神经和血液周围组织中白细胞介素-22（IL-22）的表达水平，荧光定量 PCR 检测其 mRNA 水平。
实验组 1 中小鼠 TGF-β1 表达较高，实验组 2 中 TGF-β1 表达较低，实验组 3 表达较高（p<0.05）。CatWalk 测试结果表明，实验组 1 的主要指标优于其他组，而实验组 2 的主要指标劣于其他组。根据免疫组织化学和 ELISA 结果，不同处理组小鼠 IL-22 的表达水平存在显著差异（p<0.05）。对照组[（5.05±0.15）pg/ml]中 IL-22 水平最低，实验组 1 中[（6.52±0.24）pg/ml]次之，实验组 2 中[（9.47±0.31）pg/ml]最高。
过表达 TGF-β 的人羊膜间充质干细胞可抑制异种周围神经移植后的排斥反应。
