Zhang Lihua, He Huiqing, Wang Jing, Sheng Deqiao
Department of Pediatrics of The Second People's Hospital of Yichang Yichang, Hubei Province, China.
Department of Public Health of The Second People's Hospital of Yichang Yichang, Hubei Province, China.
Int J Clin Exp Med. 2015 Feb 15;8(2):2459-64. eCollection 2015.
To investigate Cartilage glycoprotein 39 (Cgp-39) expression in peripheral blood monocytes of septic rats, and analyze the relationship between Toll-like receptor 4 (TLR4)-NF-κB signalling pathway and Cgp-39 expression.
The ligation puncture was performed to establish rat sepsis model, and ELISA was used to measure serum Cgp-39 concentration. Peripheral blood mononuclear cells was isolated and cultured for 72 h. RNA interference technology was used to inhibit TLR4 and NF-κB gene expression, and real-time PCR and Western blot were performed to detect mRNA and protein expression of TLR4 and NF-κB.
At 1 h, there was no significant differences in serum Cgp-39 concentration between sepsis group and the control group (P > 0.05), however, at 6 h, 12 h, 24 h and 48 h, serum Cgp-39 concentrations in sepsis group were significantly higher than those in the control group at the corresponding time points (P < 0.05). Compared with the control group, TLR4 mRNA and protein expression were increased significantly in sepsis group and sepsis NF-κB interference group; NF-κB mRNA and protein expression were increased significantly in sepsis group and sepsis TLR4 interference group. However, compared with sepsis group, Cgp-39 concentrations decreased significantly in either sepsis TLR4 interference group or NF-κB interference group (P < 0.05 for both).
Cgp-39 is highly expressed in peripheral blood monocytes of septic rat and TLR4-NF-κB signalling pathways may be involved in the regulation of Cgp-39 expression.
探讨软骨糖蛋白39（Cgp-39）在脓毒症大鼠外周血单核细胞中的表达情况，并分析Toll样受体4（TLR4）-核因子κB（NF-κB）信号通路与Cgp-39表达之间的关系。
采用结扎穿刺法建立大鼠脓毒症模型，酶联免疫吸附测定（ELISA）法检测血清Cgp-39浓度。分离外周血单个核细胞并培养72小时。运用RNA干扰技术抑制TLR4和NF-κB基因表达，采用实时荧光定量聚合酶链反应（real-time PCR）和蛋白质免疫印迹法（Western blot）检测TLR4和NF-κB的mRNA及蛋白表达。
1小时时，脓毒症组与对照组血清Cgp-39浓度无显著差异（P>0.05）；然而，在6小时、12小时、24小时和48小时时，脓毒症组血清Cgp-39浓度在相应时间点均显著高于对照组（P<0.05）。与对照组相比，脓毒症组和脓毒症NF-κB干扰组中TLR4的mRNA和蛋白表达显著增加；脓毒症组和脓毒症TLR4干扰组中NF-κB的mRNA和蛋白表达显著增加。但是，与脓毒症组相比，脓毒症TLR4干扰组或NF-κB干扰组中Cgp-39浓度均显著降低（两者均P<0.05）。
Cgp-39在脓毒症大鼠外周血单核细胞中高表达，TLR4-NF-κB信号通路可能参与Cgp-39表达的调控。
