Yan Xiao-wen, Li Wei-qin, Li Qiu-rong, Li Ning, Li Jie-shou
Research Institute of General Surgery, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, China.
Zhonghua Wai Ke Za Zhi. 2009 Aug 15;47(16):1257-60.
To investigate the effects of nuclear factor kappa B (NF-kappaB) on insulin signaling in skeletal muscle cells of rat with sepsis.
SD rats were randomly divided into two groups: control group and sepsis group.Sepsis model was reproduced by cecal ligation and puncture in sepsis group. At 8, 16, 24, 48 and 72 h after operation, the gastrocnemius was harvested. Conventional HE staining was used to observe the morphology of skeletal muscle cells. IRS-1 protein and tyrosine phosphorylation of IRS-1 and Ser(307) phosphorylation of IRS-1 were detected by Western Blotting and immuno-precipitation. Activities of NF-kappaB in skeletal muscle cells were detected by electrophoretic mobility shift assay.
Tyrosine phosphorylation of IRS-1 in sepsis group was significantly lower than in control group (P < 0.01), while Ser(307) phosphorylation of IRS-1 in sepsis group was significantly higher than in control group (P < 0.01). In sepsis group, NF-kappaB activity in skeletal muscle cells was significantly higher than in control group (P < 0.01). There was significant negative correlation between activity of NF-kappaB and tyrosine phosphorylation of IRS-1 (r = 0.972, P < 0.01). There was significant positive correlation between activities of NF-kappaB and Ser(307) phosphorylation of IRS-1 (r = 0.969, P < 0.01).
There is no inflammatory cell infiltrate in skeletal muscle cells with sepsis. But the activity of NF-kappaB in skeletal muscle cells is obviously enhanced, and it is closely related with disorder of insulin signaling in skeletal muscle cells of rat with sepsis.
探讨核因子κB（NF-κB）对脓毒症大鼠骨骼肌细胞胰岛素信号的影响。
将SD大鼠随机分为两组：对照组和脓毒症组。脓毒症组采用盲肠结扎穿孔法复制脓毒症模型。术后8、16、24、48和72小时，取腓肠肌。采用常规HE染色观察骨骼肌细胞形态。通过蛋白质免疫印迹法和免疫沉淀法检测胰岛素受体底物-1（IRS-1）蛋白、IRS-1酪氨酸磷酸化及IRS-1丝氨酸（Ser）307磷酸化。采用电泳迁移率变动分析检测骨骼肌细胞中NF-κB的活性。
脓毒症组IRS-1酪氨酸磷酸化水平显著低于对照组（P<0.01），而脓毒症组IRS-1丝氨酸307磷酸化水平显著高于对照组（P<0.01）。脓毒症组骨骼肌细胞中NF-κB活性显著高于对照组（P<0.01）。NF-κB活性与IRS-1酪氨酸磷酸化呈显著负相关（r = 0.972，P<0.01）。NF-κB活性与IRS-1丝氨酸307磷酸化呈显著正相关（r = 0.969，P<0.01）。
脓毒症大鼠骨骼肌细胞无炎性细胞浸润。但骨骼肌细胞中NF-κB活性明显增强，且与脓毒症大鼠骨骼肌细胞胰岛素信号紊乱密切相关。
