阻断粒细胞集落刺激因子受体减轻对乙酰氨基酚引起的小鼠肝损伤的研究

Blockade of the G-CSF receptor attenuates APAP-induced liver injury in mice

目的 探讨阻断粒细胞集落刺激因子受体(G-CSFR)减轻对乙酰氨基酚(APAP)引起的小鼠肝损伤的作用。方法 28只雄性C57BL/6小鼠随机分为空白对照组(Sham组)、APAP模型组、同型抗体(Isotype)对照组及抗G-CSFR组,每组各7只。通过腹腔注射APAP建立急性肝损伤模型。Isotype对照组、抗GCSFR组小鼠在注射APAP前24 h分别向腹腔注射BM4抗体及VR81抗体。模型建立24 h后获得样本,检测血清丙氨酸氨基转移酶(ALT)及天门冬氨酸氨基转移酶(AST)含量;HE染色观察肝组织病理学变化;酶联免疫吸附试验检测血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平;实时荧光定量PCR检测肝组织IL-6、单核细胞趋化蛋白1(MCP-1)和巨噬细胞炎症蛋2(MIP-2),补体成分5a受体(C5aR)和C-X-C趋化因子受体(CXCR2)表达情况;Western blot法测定Caspase-3蛋白的表达水平。结果 与空白对照组比较,APAP模型组小鼠血清中ALT及AST的水平均升高(均P <0.05)。与APAP模型组相比,抗G-CSFR组肝组织病变明显减轻。与APAP模型组相比,抗G-CSFR组炎症因子TNF-α、IL-6水平较低,肝组织中IL-6、MCP-1/CCL2、MIP2/CXCL2、C5aR和CXCR2表达下降差异均有统计学意义(均P<0.05)。Western blot结果显示阻断G-CSFR减少了Caspase-3蛋白的表达。结论 阻断G-CSFR可降低APAP诱导急性肝损伤的程度,可能成为治疗肝损伤的靶点。

Objective To investigate the effect of blocking granulocyte colony stimulating factor(G-CSF)receptor(G-CSFR)on liver injury in vivo,and to explore the potential of G-CSFR as a therapeutic target for alleviating liver injury.Methods Twenty-eight male C57 BL/6 mice were randomly divided into blank control group,APAP model group,iso type antibody control group and anti-G-CSFR group,with seven mice in each group.Acute liver injury model was established by intraperitoneal injection of APAP.The mice in the isotype antibody control group and antiG-CSFR group were injected with BM4 antibody and VR81 antibody respectively 24 hours before the injection of APAP.Samples were obtained 24 hours after the model was established.Serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected.HE staining was used to observe the pathological changes of liver tissue,and serum tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by enzyme-linked immunosorbent assay.Real-time fluorescence quantitative PCR was used to detect the expression of IL-6,monocyte chemoattractant protein 1(MCP-1),macrophage inflammatory protein 2(MIP2),complement component 5 a receptor(C5 aR)and C-X-C chemokine receptor 2(CXCR2)in liver tissue.Western blot was used to determine the expression level of Caspase-3 protein.Results The serum levels of ALT and AST in the APAP model group were higher those of the blank control group(all P<0.05).Compared with the APAP model group,the liver tissue lesions in the antiG-CSFR group were reduced;the anti-G-CSFR group had lower levels of TNF-αand IL-6,and lower expressions of IL-6,MCP-1/CCL2 and MIP2/CXCL2,C5 aR and CXCR2 in liver tissue,with significant differences(all P<0.05).Western blot results showed that blocking G-CSFR reduced the expression of Caspase-3 protein.Conclusions Blocking the G-CSFR can alleviate APAP-induced liver injury,and may become a target for the treatment of acute liver inj ury.