Department of Nephrology, The Affiliated Wuxi No.2 People's Hospital of Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
Department of Nephrology, The Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, Jiangsu 214023, P.R. China.
Mol Med Rep. 2021 Aug;24(2). doi: 10.3892/mmr.2021.12245. Epub 2021 Jun 29.
A humanized anti‑Toll‑like receptor 4 (TLR4) monoclonal antibody (mAb) was previously produced using phage antibody library technology, and it was found that the mAb could effectively ameliorate lipopolysaccharide (LPS)‑induced damage in macrophages. The present study investigated the protective effects exerted by the humanized anti‑TLR4 mAb against LPS‑induced acute kidney injury (AKI), as well as the underlying mechanisms. Female C57BL/6 mice were randomly divided into four groups (n=8 per group): i) Control; ii) LPS; iii) LPS + humanized anti‑TLR4 mAb (1 µg/g); and iv) LPS + humanized anti‑TLR4 mAb (10 µg/g). Serum creatinine, blood urea nitrogen, IL‑6, TNFα and IL‑1β levels were then examined, followed by renal pathology assessment, immunohistochemical staining, reverse transcription‑quantitative PCR and western blotting to assess apoptosis/survival/inflammation‑related molecules and kidney injury molecule (KIM)‑1. The humanized anti‑TLR4 mAb successfully ameliorated LPS‑induced AKI and renal pathological damage. The humanized anti‑TLR4 mAb also dose‑dependently suppressed LPS‑induced elevations in serum IL‑6, TNFα and IL‑1β, and decreased the renal expression levels of myeloid differentiation primary response 88 (MyD88), IKKα/β, IκB, p65 and KIM‑1. Compared with the LPS group, renal Bax and KIM‑1 expression levels were significantly downregulated, and Bcl‑2 expression was notably upregulated by the humanized anti‑TLR4 mAb. Moreover, the humanized anti‑TLR4 mAb also significantly decreased the protein expression levels of MyD88, phosphorylated (p)‑IKKα/β, p‑IκB and p‑p65 in the renal tissues compared with the LPS group. Therefore, the present study indicated that the anti‑inflammatory effects of the humanized anti‑TLR4 mAb against LPS‑related AKI in mice were mediated via inhibition of the TLR4/NF‑κB signaling pathway.
一种人源化抗 Toll 样受体 4（TLR4）单克隆抗体（mAb）先前已使用噬菌体抗体库技术生产，研究发现该 mAb 可有效改善脂多糖（LPS）诱导的巨噬细胞损伤。本研究探讨了人源化抗 TLR4 mAb 对 LPS 诱导的急性肾损伤（AKI）的保护作用及其潜在机制。雌性 C57BL/6 小鼠随机分为四组（每组 8 只）：i）对照组；ii）LPS 组；iii）LPS+人源化抗 TLR4 mAb（1μg/g）组；iv）LPS+人源化抗 TLR4 mAb（10μg/g）组。检测血清肌酐、血尿素氮、IL-6、TNFα 和 IL-1β 水平，然后进行肾脏病理评估、免疫组化染色、逆转录-定量 PCR 和 Western blot 检测凋亡/存活/炎症相关分子和肾损伤分子（KIM）-1。人源化抗 TLR4 mAb 成功改善了 LPS 诱导的 AKI 和肾脏病理损伤。人源化抗 TLR4 mAb 还呈剂量依赖性抑制 LPS 诱导的血清 IL-6、TNFα 和 IL-1β 升高，并降低髓样分化初级反应 88（MyD88）、IKKα/β、IκB、p65 和 KIM-1 的肾表达水平。与 LPS 组相比，人源化抗 TLR4 mAb 下调肾 Bax 和 KIM-1 表达水平，上调 Bcl-2 表达水平。此外，人源化抗 TLR4 mAb 还显著降低了 LPS 组肾组织中 MyD88、磷酸化 IKKα/β（p-IKKα/β）、p-IκB 和 p-p65 的蛋白表达水平。因此，本研究表明，人源化抗 TLR4 mAb 对 LPS 相关 AKI 的抗炎作用是通过抑制 TLR4/NF-κB 信号通路介导的。
