Department of Nephrology, First Affiliated Hospital of ChongQing Medical University, ChongQing, 400016, China.
Inflammation. 2012 Feb;35(1):388-96. doi: 10.1007/s10753-011-9374-9.
Diabetic nephropathy (DN) is a major cause of type 2 diabetes mellitus (T2DM) mortality. Innate immunity has been shown to be closely associated with the occurrence and progression of T2DM-associated complications. In this study, we investigated the expression of Toll-like receptor 4 (TLR4) and CD14(+)CD16(+) monocytes in patients with T2DM and DN patients with uremia and TLR4 response to lipopolysaccharide (LPS), and to further explore the potential effects of inflammatory immune response in T2DM and DN uremia. Thirty DN patients with uremia, 28 T2DM patients, and 20 healthy volunteers were enrolled for the determination of CD14(+)CD16(+) fluorescence intensity and TLR4 expression on monocytes by using peripheral blood flow cytometry. Serum C-reactive protein (CRP) level was determined by using the immunoturbidimetry. Peripheral blood mononuclear cells (PBMCs) were isolated and stimulated with LPS for 24 h. monocytes were collected to detect NF-κB p65 and phosphorylated STAT5(p-STAT5) expressions by using Western blotting. Supernatants were sampled for the determination of interleukin-6 (IL-6) concentration by using ELISA. Compared to normal control, T2DM patients and DN uremic patients had a significantly higher CD14(+)CD16(+) fluorescence intensity, TLR4 expression, serum IL-6 and CRP level, whilst these biomarkers were more upregulated in DN uremic patients than in T2DM patients. Following the exposure to LPS, PBMCs showed a significant upregulation in NF-κB-p65 and p-STAT5 expression and a remarked increase in Supernatants IL-6 level, in a positive correlation with disease severity. Our results suggest that the disturbance in proinflammatory CD14(+)CD16(+) monocytes occurs in T2DM and DN uremic patients. Such immunological dysfunction may be related to the activation of TLR4/NF-κB and STAT5 signaling pathways underlying the immune abnormalities of CD14(+)CD16(+) monocytes.
糖尿病肾病 (DN) 是 2 型糖尿病 (T2DM) 患者死亡的主要原因。先天免疫与 T2DM 相关并发症的发生和发展密切相关。在这项研究中，我们研究了 Toll 样受体 4 (TLR4) 和 CD14(+)CD16(+)单核细胞在 T2DM 和 DN 尿毒症患者中的表达，以及 TLR4 对脂多糖 (LPS) 的反应，并进一步探讨了炎症免疫反应在 T2DM 和 DN 尿毒症中的潜在作用。30 例 DN 尿毒症患者、28 例 T2DM 患者和 20 例健康志愿者被纳入本研究，通过外周血流式细胞术测定单核细胞上的 CD14(+)CD16(+)荧光强度和 TLR4 表达。采用免疫比浊法测定血清 C 反应蛋白 (CRP) 水平。分离外周血单个核细胞 (PBMC)，用 LPS 刺激 24 h。收集单核细胞，用 Western blot 检测 NF-κB p65 和磷酸化 STAT5(p-STAT5)的表达。采用 ELISA 法测定上清液中白细胞介素-6 (IL-6)的浓度。与正常对照组相比，T2DM 患者和 DN 尿毒症患者的 CD14(+)CD16(+)荧光强度、TLR4 表达、血清 IL-6 和 CRP 水平明显升高，而 DN 尿毒症患者的这些标志物水平明显高于 T2DM 患者。在 LPS 刺激后，PBMC 中 NF-κB-p65 和 p-STAT5 的表达明显上调，上清液中 IL-6 水平明显升高，与疾病严重程度呈正相关。我们的结果表明，在 T2DM 和 DN 尿毒症患者中存在促炎 CD14(+)CD16(+)单核细胞紊乱。这种免疫功能障碍可能与 TLR4/NF-κB 和 STAT5 信号通路的激活有关，该通路是 CD14(+)CD16(+)单核细胞免疫异常的基础。
