目的 研究细胞因子信号抑制物1(suppressor of cytokine signaling 1，SOCS1)基因在高糖诱导的小鼠腹膜间皮细胞上皮间质转化(epithelial mesenchymal transformation，EMT)中的作用及机制。 方法 培养小鼠腹膜间皮细胞并分组，对照组用普通培养基处理，1.5%组、2.5%组、4.25%组分别加入相应浓度的D-葡萄糖；2.5%+阴性对照(negative control，NC)质粒组2.5%+SOCS1质粒组在含有2.5% D-葡萄糖的培养基中分别转染NC质粒和SOCS1质粒。处理24小时后检测细胞增殖，迁移和侵袭数目，SOCS1、E-钙粘蛋白(cadherin)、N-钙粘蛋白(N-cadherin)、α-平滑肌肌动蛋白(α-smooth muscle actin，α-SMA)的mRNA表达水平，Janus激酶(Janus kinase，JAK)1/磷酸化信号转导和转录激活因子(signal transduction and activator of transcription，STAT)1、JAK2/STAT3通路。 结果 4.25%组小鼠腹膜间皮细胞的增殖水平低于对照组(t=5.051、P=0.002)；1.5%组、2.5%组、4.25%组小鼠腹膜间皮细胞中E-cadherin、SOCS1的mRNA表达水平低于对照组(t=2.774、7.310、9.7813，P=0.032、＜0.001； t=2.544、6.996、10.733，P=0.044、＜0.001)，N-cadherin(t=2.825、5.121、7.207，P=0.030、＜0.001)、 α-SMA(t=2.527、4.807、6.950，P=0.045、0.003、＜0.001)的表达水平高于对照组；2.5%+SOCS1质粒组小鼠腹膜间皮细胞中JAK1/STAT1和JAK2/STAT3信号通路受抑制，SOCS1的mRNA表达水平和蛋白表达水平、E-cadherin的mRNA表达水平高于2.5%组(t=9.435、9.761、7.690，均P＜0.001)、2.5%+NC质粒组 (t=9.737、9.138、7.132，均P＜0.001)，细胞迁移和侵袭数目、N-cadherin、α-SMA的mRNA表达水平水平低于2.5%组、2.5%+NC质粒组(t=7.350、8.456，P＜0.001；t=8.562、7.697，P＜0.001；t=4.574、4.865， P=0.004、0.003；t=3.467、3.036，P=0.013、0.023)。 结论 过表达SOCS1基因抑制高糖诱导小鼠腹膜间皮细胞EMT，这一作用与抑制JAK1/STAT1和JAK2/STAT3信号通路相关。
Objective To investigate the role and mechanism of the suppressor of cytokine signaling 1 (SOCS1) gene in high glucose-induced epithelial-mesenchymal transformation (EMT) of peritoneal mesothelial cells in mice. Methods Mouse peritoneal mesothelial cells were cultured and divided into 6 groups: the control group was treated with routine medium; cells cultured in the medium containing 1.5% (83 mmol/L), 2.5% (139 mmol/L) and 4.25% (236 mmol/L) D-glucose formed the 1.5% group, 2.5% group and 4.25% group; cells transfected with negative control plasmid and cultured in the medium containing 2.5% glucose formed the 2.5%+NC plasmid group; cells transfected with SOCS1 plasmid and cultured in the medium containing 2.5% glucose formed the 2.5%+SOCS1 plasmid group. After the treatment for 24h, proliferation, migration and invasion number of the cells, mRNA expression levels of SOCS1, E-cadherin, N-cadherin and α-SMA, and JAK1/STAT1 and JAK2/STAT3 signaling pathways were detected. Results In 4.25% group, the proliferation level of peritoneal mesothelial cells was lower than that in control group (t=5.051, P=0.002). In 1.5%, 2.5% and 4.25% groups, the mRNA expression levels of E-cadherin and SOCS1 in peritoneal mesothelial cells were lower than those in control group (E-cadherin mRNA: t=2.774, 7.310 and 9.7813; P=0.032, ＜0.001 and ＜0.001. SOCS1 mRNA: t=2.544, 6.996 and 10.733; P=0.044, ＜0.001 and ＜0.001), while mRNA expression levels of N-cadherin and α-SMA in peritoneal mesothelial cells were higher than those in control group (N-cadherin mRNA: t=2.825, 5.121 and 7.207; P=0.030, ＜0.001 and ＜0.001. α-SMA mRNA: t=2.527, 4.807 and 6.950; P=0.045, 0.003 and ＜0.001). In 2.5%+SOCS1 plasmid group, JAK1/STAT1 and JAK2/STAT3 signaling pathways in peritoneal mesothelium cells were inhibited, the mRNA and protein expression levels of SOCS1 and the mRNA expression level of E-cadherin were higher than those in 2.5% group and 2.5%+NC plasmid group (SOCS1 mRNA: t=9.435 and 9.737, P＜0.001; SOCS1 protein: t=9.761 and 9.138, P＜0.001; E-cadherin mRNA: t=7.690 and 7.132, P＜0.001), and the number of cell migration and invasion and the mRNA expression levels of N-cadherin and α-SMA were lower than those in 2.5% group and 2.5%+NC plasmid group (t=7.350 and 8.456, P＜0.001; t=8.562 and 7.697, P＜0.001; t=4.574 and 4.865, P=0.004 and 0.003; t=3.467 and 3.036, P=0.013, 0.023). Conclusion Overexpression of SOCS1 gene inhibits high glucose-induced EMT of peritoneal mesothelial cells in mice, which is related to the inhibition of JAK1/STAT1 and JAK2/STAT3 signaling pathways.
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