Han Mei, Li Ying, Wen Di, Liu Maodong, Ma Yuteng, Cong Bin
Department of Emergency, The Second Hospital of Hebei Medical University, Shijiazhuang, China.
Department of Nephropathy, The Third Hospital of Hebei Medical University, Shijiazhuang, 050051, China.
BMC Nephrol. 2018 Jul 6;19(1):168. doi: 10.1186/s12882-018-0977-3.
We sought to confirm that neutrophil gelatinase-associated lipocalin (NGAL) protects against apoptosis during endotoxemia.
Endotoxemia was induced in rats with lipopolysaccharide (LPS; 3.5 mg/kg) and serum creatinine (SCr), urinary NGAL (uNGAL), renal histopathology confirmed acute kidney injury (AKI). Renal caspase 3 and NGAL were assayed with immunohistochemistry 6 h later. A HK-2 cell model was used in which NGAL and caspase 3 mRNA were evaluated by qRT-PCR within 6 h after LPS (50 μM) treatment, and correlations were studied. NGAL and caspase 3 mRNA expression were measured after delivering NGAL siRNA in HK-2 cells and apoptosis was measured with TUNEL and flow cytometry.
SCr and uNGAL were significantly increased after LPS treatment and renal morphology data indicated AKI and renal tubular epithelial cell apoptosis. Caspase 3 and NGAL were predominantly expressed in the tubular epithelial cells and there was a correlation between caspase 3 and NGAL protein (r = 0.663, p = 0.01). In vitro, there was a strong correlation between caspase 3 and NGAL mRNA in LPS-injured HK-2 cells within 24 h (r = 0.448, p < 0.05). Suppressing the NGAL gene in HK-2 cells increased caspase 3 mRNA 4.5-fold and apoptosis increased 1.5-fold after LPS treatment.
NGAL is associated with caspase 3 in renal tubular cells with endotoxin-induced kidney injury, and may regulate its expression and inhibit apoptosis.
我们试图证实中性粒细胞明胶酶相关脂质运载蛋白（NGAL）在内毒素血症期间可防止细胞凋亡。
用脂多糖（LPS；3.5毫克/千克）诱导大鼠发生内毒素血症，检测血清肌酐（SCr）、尿NGAL（uNGAL），肾组织病理学检查证实为急性肾损伤（AKI）。6小时后用免疫组织化学法检测肾组织中的半胱天冬酶3和NGAL。使用HK-2细胞模型，在LPS（50微摩尔）处理后6小时内通过qRT-PCR评估NGAL和半胱天冬酶3 mRNA，并研究其相关性。在HK-2细胞中导入NGAL siRNA后检测NGAL和半胱天冬酶3 mRNA表达，并用TUNEL和流式细胞术检测细胞凋亡。
LPS处理后SCr和uNGAL显著升高，肾脏形态学数据表明存在AKI和肾小管上皮细胞凋亡。半胱天冬酶3和NGAL主要在肾小管上皮细胞中表达，半胱天冬酶3和NGAL蛋白之间存在相关性（r = 0.663，p = 0.01）。在体外，LPS损伤的HK-2细胞在24小时内半胱天冬酶3和NGAL mRNA之间存在强相关性（r = 0.448，p < 0.05）。在HK-2细胞中抑制NGAL基因可使LPS处理后半胱天冬酶3 mRNA增加4.5倍，细胞凋亡增加1.5倍。
在内毒素诱导的肾损伤中，NGAL与肾小管细胞中的半胱天冬酶3相关，可能调节其表达并抑制细胞凋亡。
