细胞因子信号转导抑制蛋白-1对高糖环境下肾小球系膜细胞生长的影响

Effect of Cytokine Signal Transduction Inhibitor-1 on the Growth of Mesangial Cells in High Glucose Environment

目的探讨细胞因子信号转导抑制蛋白-1(SOCS-1)及高糖培养条件对肾小球系膜细胞(HMC)生长的影响。方法体外培养人肾小球细胞,应用脂质体2000转染SOCS-1 siRNA表达质粒及SOCS-1无意义si RNA表达质粒。分为:对照组为甘露醇组;高糖模型组;高糖模型组+SOCS-1 siRNA干扰;高糖模型组+无意义si RNA,高糖模型组用DMEM培养液中加入葡萄糖,终浓度为30 mmol/L;流式检测细胞凋亡。Western印迹和荧光定量PCR检测系膜细胞信号转导和转录活化因子3(STAT3)和核转录因子Kappa B(NF-κB)的表达。结果高糖+SOCS-1干扰组细胞凋亡率1.01%,对照组1.52%,高糖模型组1.95%,SOCS-1 siRNA干扰组细胞凋亡率下调,差异有统计学意义(P<0.05)。高糖模型组相对于对照组,Western Blot检测显示NF-kB水平表达上调[(1.21±0.08) vs (0.47±0.04),t=9.49,P<0.01];STAT3表达上调[(1.37±0.12) vs (0.54±0.06),t=10.64,P<0.01];SOCS-1 siRNA干扰组相对于无意义si RNA组,NF-kB水平表达上调[(0.80±0.08) vs (0.32±0.02),t=10,P<0.01]、STAT3表达上调[(0.87±0.10) vs (0.32±0.02),t=9.17,P<0.01]。(RT-PCR检测提示高糖模型组相对于对照组,NF-kB[(1.15±0.15)vs (0.80±0.16),t=2.9,P<0.05]、STAT3[(0.75±0.02)vs (0.46±0.09),t=5.686,P<0.01]表达上调;SOCS-1 siRNA干扰组相对于无意义si RNA组,NF-kB[(1.15±0.17)vs (0.80±0.09),t=3.18,P<0.05]、STAT3[(0.88±0.06)vs (0.40±0.01),t=13.7,P<0.01]表达上调。结论高糖环境会上调HMC的JAK/STAT信号通路,促进HMC的生长;干扰SOCS-1会上调JAK/STAT信号通路,并促进NF-κB的表达,降低细胞凋亡率,促进高糖环境下HMC的生长。

Objective To investigate the effects of cytokine signal transduction inhibitor-1 (SOCS-1) and high glucose culture conditions on the growth of mesangial cells (HMC). Methods Human glomerular cells were cultured in vitro, and SOCS-1 siRNA expression plasmid and SOCS-1 nonsense siRNA expression plasmid were transfected with liposome 2000, and were divided into: control group as the mannitol group;high glucose model group;high glucose model group + SOCS-1 siRNA interference;high glucose model group + meaningless siRNA, high glucose model group with DMEM culture medium added glucose, the final concentration was 30 mmol/L;flow detection of apoptosis;Western blot and real-time PCR were used to detect the expression of mesangial cell signal transduction and transcriptional activator 3 (STAT3) and nuclear transcription factor KappaB (NF-κB). Results The apoptotic rate was 1.01% in the high glucose+SOCS-1 interference group, 1.52% in the control group, and 1.95% in the high glucose model group. The apoptosis rate of the SOCS-1 siRNA interference group was down-regulated, the difference was statistically significant (P<0.05). Compared with the control group, Western Blot showed that the expression of NF-kB was up-regulated [(1.21±0.08) vs (0.47±0.04), t=9.49, P<0.01];STAT3 expression was up-regulated[(1.37±0.12) vs (0.54±0.06), t=10.64, P<0.01];SOC-kB level was up-regulated in SOCS-1 siRNA interference group compared with non-meaningful siRNA group [(0.80±0.08) vs (0.32±0.02), t=10, P<0.01], STAT3 expression was up-regulated [(0.87±0.10) vs (0.32±0.02), t=9.17, P<0.01]. RT-PCR detection indicated that the high glucose model group was compared with the control group, NF-kB [(1.15±0.15) vs (0.80±0.16), t=2.9, P<0.05], STAT3[(0.75±0.02) vs (0.46±0.09), t=5.686, P<0.01] expression up-regulation;SOCS-1 siRNA interference group vs. nonsense siRNA group, NF-kB[(1.15±0.17) vs (0.80±0.09), t=3.18, P<0.05], STAT3 [(0.88±0.06) vs (0.40±0.01), t=13.7, P<0.01] expression was up-regulated. Conclusion The high glucose environment up-regulates the JAK/STAT signaling pathway of HMC and promotes the growth of HMC. Interfering with SOCS-1 up-regulates JAK/STAT signaling pathway, promotes the expression of NF-κB, decreases the apoptosis rate, and promotes the growth of HMC in high glucose environment.