肥胖降低乙型肝炎患者干扰素治疗敏感性的机制研究

The mechanism of obesity reducing the sensitivity of interferon therapy in hepatitis B patients

目的探讨乙型肝炎(以下简称乙肝)患者干扰素(IFN)治疗效果与机体肥胖程度的关系,以及肥胖致IFN治疗乙肝不敏感的分子机制与炎症细胞因子的关系。方法以肥胖作为唯一影响因素,通过文献检索,筛选并提取文献数据,经过RevMan软件作图分析,分析肥胖和IFN治疗乙肝敏感性的关系。选取2017-2021年于该院行IFN治疗的乙肝患者91例为研究对象,根据体质量指数(BMI)将其分为对照组(BMI≤25 kg/m^(2),61例)和肥胖组(BMI≥28 kg/m^(2),30例)。跟踪IFN治疗48周,比较两组谷氨酸氨基转移酶(ALT)和乙肝表面抗原(HBsAg)水平。收集不同肥胖程度的健康查体人群血浆,分为A(BMI≤25 kg/m^(2))、B(BMI≥28 kg/m^(2))两组,借助流式细胞术检测炎症细胞因子[白细胞介素(IL)-2、IL-4、IL-6、IL-10、IL-12p70、IL-17、肿瘤坏死因子(TNF)-α、IFN-γ]水平,寻找A、B两组中差异有统计学意义的炎症细胞因子。体外培养稳定转染人HBV基因组的HepG 2.2.15细胞,IFNα2b与筛选出的差异细胞因子单独或联合作用。收集细胞培养上清液,检测HBsAg、乙型肝炎e抗原(HBeAg)、HBV-DNA水平,并检测IFN信号转导通路JAK-STAT和抗病毒蛋白IFITM1、MX1、OAS1、PKR水平。结果与对照组比较,肥胖组ALT、HBsAg水平升高(P<0.05)。B组IL-2、IL-6、IL-10、IL-17水平明显高于A组,差异有统计学意义(P<0.05)。IFNα2b单独或者与IL联合作用下,JAK1、STAT1、STAT2发生不同程度的磷酸化,JAK-STAT信号通路被激活,其中IL-6、IL-10降低JAK1-STAT磷酸化的程度尤其明显。IFN单独作用下,抗病毒蛋白IFITM1、MX1、OAS1、PKR水平升高,与IL-6、IL-10联合作用下,上述指标水平升高更加明显(P<0.05)。结论肥胖降低乙肝患者IFN治疗的敏感性,但是其机制并不与IL-2、IL-6、IL-10、IL-17水平升高直接有关。IL-2、IL-6、IL-10、IL-17可以上调抗病毒蛋白IFITM1、MX1、OAS1、PKR水平。

Objective To investigate the relationship between the effect of interferon(IFN)treatment and the degree of obesity in patients with chronic hepatitis B,as well as the relationship between the molecular mechanism of obesity-induced insensitivity to IFN therapy and inflammatory cytokines.Methods Taking obesity as the only influencing factor,through literature search,screened and extracted literature data,and analyzed the relationship between obesity and IFN treatment susceptibility to hepatitis B through RevMan software.A total of 91 hepatitis B patients who received IFN therapy in this hospital from 2017 to 2021 were selected as the research objects,and they were divided into control group(BMI≤25 kg/m^(2),61 cases)and obesity group(BMI≥28 kg/m^(2),30 cases)according to body mass index(BMI).The IFN treatment was followed up for 48 weeks,and the levels of glutamate aminotransferase(ALT)and hepatitis B surface antigen(HBsAg)were compared between the two groups.The plasma of healthy people with different degrees of obesity was collected and divided into two groups:A group(BMI≤25 kg/m^(2),61 cases)and B group(BMI≥28 kg/m^(2)).Flow cytometry was used to detect inflammatory cytokines[interleukin(IL)-2,IL-4,IL-6,IL-10,IL-12p70,IL-17,tumor necrosis factor(TNF)-α,IFN-γ]levels,the inflammatory cytokines with statistically significant differences between groups A and B were searched.HepG 2.2.15 cells stably transfected with human HBV genome were cultured in vitro,and IFNα2b interacted with the selected differential cytokines alone or in combination.Cell culture supernatants were collected,the levels of HBsAg,hepatitis Be antigen(HBeAg),HBV-DNA,IFN signal transduction pathway JAK-STAT and antiviral proteins IFITM1,MX1,OAS1,and PKR were detected.Results Compared with the control group,the levels of ALT and HBsAg in the obese group were increased(P<0.05).The levels of IL-2,IL-6,IL-10 and IL-17 in group B were significantly higher than those in group A,and the differences were statistically significant(P<0.05).Under the action of IFNα2b alone or in combination with IL,JAK1,STAT1,and STAT2 were phosphorylated to different degrees,and the JAK-STAT signaling pathway was activated.Among them,IL-6 and IL-10 significantly reduced the phosphorylation of JAK1-STAT.The levels of antiviral proteins IFITM1,MX1,OAS1,and PKR increased under the action of IFN alone,and the levels of the above indexes increased more significantly under the combined action of IL-6,IL-10(P<0.05).Conclusion Obesity reduces the sensitivity of IFN therapy in hepatitis B patients,but the mechanism is not directly related to the elevated levels of IL-2,IL-6,IL-10,and IL-17.IL-2,IL-6,IL-10 and IL-17 can up-regulate the levels of antiviral proteins IFITM1,MX1,OAS1 and PKR.