HIF-1α、Treg和Th17在非酒精性脂肪性肝病患者外周血中的表达及其临床意义

目的 研究非酒精性脂肪性肝病(nonalcoholic fatty liver disease，NAFLD)患者外周血中缺氧诱导因子-1α(HIF-1α)的表达以及T淋巴细胞中亚群Treg和Th17细胞的比例，探讨三者与非酒精性脂肪性肝病发病机制的关系。 方法 收集了自2015年10月—2016年4月在湖州市中心医院消化内科确诊的非酒精性脂肪性肝病患者39名，以及健康志愿者20名。采用流式细胞仪采集分析受试者CD4+ T淋巴细胞内CD4+/CD25+/Foxp3+的Treg细胞和CD4+/IL1-17+的Th17细胞的比例。使用酶联免疫吸附法测定受试者血清HIF-1α的表达情况。比较患者与健康者血清HIF-1α的表达水平以及Th17和Treg的细胞比例。同时亦比较肥胖(BMI≥28 kg/m2)与非肥胖患者(BMI<28 kg/m2)上述三项指标的差异。采用独立样本t检验和χ2检验进行统计学分析。 结果 非酒精性脂肪性肝病患者血清HIF-1α的表达明显高于对照组(t=11.228，P<0.001)，Th17和Treg细胞在T淋巴细胞中的比例也明显升高(t=11.313,P<0.001和t=8.737,P<0.001)并且Th17/Treg的比例也有显著提高(t=4.055，P<0.001)。非肥胖NAFLD患者的血清HIF-1α表达水平明显高于肥胖患者(t=2.035，P<0.05)，Th17和Treg占比也有所升高(t=2.066,P<0.05和t=2.089，P<0.05)，但Th17/Treg比值差异无统计学意义(t=0.048，P>0.05)。 结论 非酒精性脂肪性肝病患者外周血中的HIF-1α表达较高，Th17/Treg平衡也出现失衡。HIF-1α和Th17/Treg平衡都可能参与了非酒精性脂肪性肝病的发生和发展，且与胰岛素抵抗存在一定的相互作用，为该病的治疗提供了新的潜在靶点。 

Expression of HIF-1α,Treg and Th17 in the peripheral blood of patients with nonalcoholic fatty liver disease

Objective To study the expression of HIF-1αin the peripheral blood and the ratio of Treg and Th17 cells in T cells in patients with nonalcoholic fatty liver disease(NAFLD),and to explore the relationship between those and pathology of NAFLD. Methods A total of 39 patients who were diagnosed as nonalcoholic fatty liver disease in our hospital from October,2015 to April,2016 and 20 healthy volunteers were collected in this study.Flow cytometry were used to measure and analyze the portion of CD4+/CD25+/Foxp3+ Treg cells and CD4+/IL17+ Th17 cells in total CD4+T lymphocytes.Enzyme linked immunosorbent assay(ELISA) were employed to test the expression level of HIF-1α.Expression of HIF-1α and the portion of Treg cells and Th17 cells were compared between two groups.They were also compared between NAFLD patients in the fat group (BMI ≥ 28 kg/m2) and those in the non-fat group(BMI<28 kg/m2).Student's t test and chi-square test were employed in the statistics. Results NAFLD patients had a much higher expression level of HIF-1α (t=11.284,P<0.001).The portion of Treg and Th17 cells in total CD4+T lymphocytes were significantly higher in NAFLD group than in control group (t=11.313,P<0.001 and t=8.737,P<0.001) and the ratio of Th17/Treg was also higher (t=4.055,P<0.001).Non-fat NAFLD patients expressed obviously higher level of HIF-1α than fat NAFLD patients(t=2.035,P<0.05).The portion of Treg and Th17 also increased in non-fat NAFLD patients (t=2.066,P<0.05 and t=2.089,P<0.05) but no significant difference of the ratio of Th17/Treg was observed (t=0.048,P>0.05). Conclusion Patients with NAFLD has higher expression level of HIF-1α and imbalance of Th17/Treg.HIF-1α and balance of Th17/Treg could be involved in the onset and development of NAFLD,and may have relationship with insulin resistance to some extent.