摘要
目的探讨5-脂氧化酶(5-LO)通路在非酒精性脂肪性肝炎(NASH)的作用。方法将4周龄ApoE-/-雄性小鼠16只随机分为两组:对照组(普通饲料)、NASH模型组(高脂高胆固醇饲料)。连续喂养12周后,肝组织油红染色观察脂肪变,采用HE染色进行NAFLD活动度评分(NAFLD activity score,NAS)。荧光定量RT-PCR检测5-LO、BLT1等mRNA表达,Western Bot法检测5-LO蛋白,酶联免疫法检测肝脏白三烯B4(leukotriene B4,LTB4)。结果高脂高胆固醇饮食12周能诱导ApoE-/-小鼠出现NASH表现,建模成功。NASH模型组较对照组肝组织5-LO mRNA及蛋白表达明显升高(P<0.01),肝匀浆LTB4水平[(71.09±18.23)pg/mL vs(46.71±11.71)pg/mL]升高(P<0.01),其受体BLT1 mRNA水平显著升高(P<0.01),中性粒细胞趋化因子和相应受体MIP2、CXCR2以及黏附分子ICAM1、VCAM1 mRNA的表达均较对照组明显升高(P<0.01)。结论 5-LO炎症通路通过上调中性粒细胞粘附浸润相关因子,诱导中性粒细胞浸润,促进非酒精性脂肪性肝炎进展。
Objective To investigate the effects of 5-lipoxygenase pathway in nonalcoholic steatohepatitis （NASH）. Methods The 16 ApoE-/- mice with the age of 4 weeks old were randomly assigned into two groups, control group （normal feed） and NASH model group（high fat and high cholesterol feed）. After 12 weeks of continuous feeding, fatty degeneration was observed by oil red staining and NAFLD activity score（NAS） was measured by HE staining. The mRNA expression of 5-LO and BLT1 was detected by fluorescent quantitative RT-PCR. 5-LO protein was detected by West- ern Blot. Leukotriene B4 （LTB4） was detected by enzyme-linked immunosorbent assay. Results High-fat and high- cholesterol diet for 12 weeks were able to induce the occurrence of NASH in ApoE-/- mice, with successful modeling. The expression of liver tissue 5-LO mRNA and the expression of protein in NASH model group was significantly higher than those in the control group （P〈0.01）. The level of LTB4 in liver homogenate was increased [（71.09_＋18.23） pg/mL vs （46.71_＋11.71） pg/mL, P〈0.01], the level of BLT1 mRNA was significantly increased in its receptor （P〈0.01）, and the expression of neutrophil chemotaxis and corresponding receptors of MIP2, CXCR2 as well as adhesion molecules of ICAM1 and VCAM1 mRNA were significantly higher than those in the control group（P〈0.01）. Conclusion 5-LO in- flammatory pathways can induce neutrophil infiltration and promote the progression of nonalcoholic steatohepatitis by up-regulating neutrophil adhesion and infiltration-related factors.
出处
《中国现代医生》
2016年第28期35-38,共4页
China Modern Doctor
基金
浙江省自然科学基金(LY14H070004)
浙江省杭州市科技局项目(20140633B09)
