Liao Wan-Qin, Qi Ya-Lei, Wang Lin, Dong Xiao-Ming, Xu Tao, Ding Chao-Dong, Liu Rui, Liang Wei-Cheng, Lu Li-Ting, Li He, Li Wen-Feng, Luo Guang-Bin, Lu Xin-Cheng
Wan-Qin Liao, Ya-Lei Qi, Lin Wang, Xiao-Ming Dong, Tao Xu, Chao-Dong Ding, Rui Liu, Wei-Cheng Liang, Li-Ting Lu, Xin-Cheng Lu, School of Basic Medical Science, Wenzhou Medical University, Chashan Campus, Chashan University Town, Wenzhou 325027, Zhejiang Province, China.
World J Gastroenterol. 2015 Sep 28;21(36):10375-84. doi: 10.3748/wjg.v21.i36.10375.
To investigate the effects of Recql5 deficiency on liver injury induced by lipopolysaccharide/D-galactosamine (LPS/D-Gal).
Liver injury was induced in wild type (WT) or Recql5-deficient mice using LPS/D-Gal, and assessed by histological, serum transaminases, and mortality analyses. Hepatocellular apoptosis was quantified by transferase dUTP nick end labeling assay and Western blot analysis of cleaved caspase-3. Liver inflammatory chemokine and cytochrome P450 expression was analyzed by quantitative reverse transcription-PCR. Neutrophil infiltration was evaluated by myeloperoxidase activity. Expression and phosphorylation of ERK, JNK, p65, and H2A.X was determined by Western blot. Oxidative stress was evaluated by measuring malondialdehyde production and nitric oxide synthase, superoxide dismutase, glutathione peroxidase, catalase, and glutathione reductase activity.
Following LPS/D-Gal exposure, Recql5-deficient mice exhibited enhanced liver injury, as evidenced by more severe hepatic hemorrhage, higher serum aspartate transaminase and alanine transaminase levels, and lower survival rate. As compared to WT mice, Recql5-deficient mice showed an increased number of apoptotic hepatocytes and higher cleaved caspase-3 levels. Recql5-deficient mice exhibited increased DNA damage, as evidenced by increased γ-H2A.X levels. Inflammatory cytokine levels, neutrophil infiltration, and ERK phosphorylation were also significantly increased in the knockout mice. Additionally, Recql5-deficient mice exhibited increased malondialdehyde production and elevated inducible nitric oxide synthase, superoxide dismutase, glutathione peroxidase, catalase, and glutathione reductase activity, indicative of enhanced oxidative stress. Moreover, CYP450 expression was significantly downregulated in Recql5-deficient mice after LPS/D-Gal treatment.
Recql5 protects the liver against LPS/D-Gal-induced injury through suppression of hepatocyte apoptosis and oxidative stress and modulation of CYP450 expression.
研究Recql5缺陷对脂多糖/ D-半乳糖胺（LPS / D-Gal）诱导的肝损伤的影响。
使用LPS / D-Gal诱导野生型（WT）或Recql5缺陷型小鼠发生肝损伤，并通过组织学、血清转氨酶和死亡率分析进行评估。通过转移酶dUTP缺口末端标记法和裂解的caspase-3的蛋白质印迹分析对肝细胞凋亡进行定量。通过定量逆转录PCR分析肝脏炎性趋化因子和细胞色素P450的表达。通过髓过氧化物酶活性评估中性粒细胞浸润。通过蛋白质印迹法测定ERK、JNK、p65和H2A.X的表达及磷酸化。通过测量丙二醛生成量以及一氧化氮合酶、超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶和谷胱甘肽还原酶活性来评估氧化应激。
在LPS / D-Gal暴露后，Recql5缺陷型小鼠表现出肝损伤加重，表现为更严重的肝出血、更高的血清天冬氨酸转氨酶和丙氨酸转氨酶水平以及更低的存活率。与WT小鼠相比，Recql5缺陷型小鼠的凋亡肝细胞数量增加，裂解的caspase-3水平更高。Recql5缺陷型小鼠表现出DNA损伤增加，γ-H2A.X水平升高证明了这一点。敲除小鼠的炎性细胞因子水平、中性粒细胞浸润和ERK磷酸化也显著增加。此外，Recql5缺陷型小鼠的丙二醛生成量增加，诱导型一氧化氮合酶、超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶和谷胱甘肽还原酶活性升高，表明氧化应激增强。此外，LPS / D-Gal处理后，Recql5缺陷型小鼠的CYP450表达显著下调。
Recql5通过抑制肝细胞凋亡和氧化应激以及调节CYP450表达来保护肝脏免受LPS / D-Gal诱导的损伤。
