HBV相关慢加亚急性肝衰竭患者外周血辅助性T淋巴细胞及相关炎症因子的表达和临床意义的初步探讨

发布时间：2018-05-09 20:25

本文选题：乙型肝炎病毒 + 慢加亚急性肝衰竭　；参考：《苏州大学》2013年硕士论文

【摘要】：背景在我国，慢性乙型病毒性肝炎(CHB)是引起慢加亚急性肝衰竭(SACLF)的主要病因，有报道称CHB占SACLF所有病因的80%。目前认为，HBV感染患者病情进展和转归受诸多因素影响，其中宿主的免疫功能发挥了重要作用，细胞因子在调节免疫应答中扮演了关键角色。辅助性T细胞（Th细胞）根据分泌的细胞因子不同分为：Th0、Th1、Th2、Th3，目前研究最多的是Th1和Th2两个亚型。研究认为在病毒性肝炎感染中，Th1细胞有利于病毒的清除，而Th2细胞主要与组织损伤及慢性化有关。最近，CD4+Th细胞家族新型成员-Th17细胞亚群的出现改写了经典的Th1/Th2细胞反应模式。该细胞以高分泌IL-17为特征，具有很强的促炎症作用。2005年Kim等人发现了IL-32，，该因子可由T细胞、NK细胞、单核细胞、上皮细胞产生，进一步研究表明其有促炎症因子的特性。IL-10是机体一种强有力的免疫和炎症抑制因子，由Th2细胞、巨噬细胞、树突状细胞（Dendritic cells，DC）和B细胞等产生，它可以减低T细胞活性，从而降低免疫反应的效应，导致机体清除病毒能力减弱。上述细胞及细胞因子在CHB中的变化已有报道，但其是否参与了HBV相关慢加亚急性肝衰竭（HBV-SACLF）的发病过程以及在肝衰竭病程中的动态变化趋势、与ALT的相关性研究未见报道。目的探讨HBV-SACLF患者外周血Th1、Th2、Th17及血清中IL-32、IL-10的动态变化趋势及其临床意义。方法采用流式细胞技术检测38例HBV-SACLF患者、46例肝衰竭前期患者、20例CHB患者、20例健康体检者（对照组），外周全血标本中Th1、Th2、Th17的表达水平，比较四组之间上述指标的差异；动态观察HBV-SACLF患者早、中、晚期Th1、Th2、Th17的变化趋势；比较感染组与非感染组、存活组与非存活组Th1、Th2、Th17表达水平的变化；分析Th1、Th2、Th17与ALT的相关性。同时采用ELISA检测上述四组血清中IL-32、IL-10的表达水平，并观察IL-32、IL-10在HBV-SACLF患者早、中、晚期的动态变化。结果1.HBV-SACLF组、肝衰竭前期组、CHB组、对照组Th1、Th2、Th17的表达水平分别为：①T h1：（22.58±3.86）%、（20.80±3.32）%、（5.23±1.51）%、（6.45±1.27）%；Th1比例在HBV-SACLF组与CHB组及对照组比较，均明显升高，差异具有显著性（P_2=0.04，P3=0.032）。②Th2：（1.73±0.64）%、（1.03±0.34）%、（6.15±0.97）%、（1.98±0.46）%；Th2比例在HBV-SACLF组与CHB组及对照组比较，均明显下降，差异具有显著性（P_2=0.007，P3=0.041）。③T h17：（3.48±0.80）%、（3.61±1.20）%、（1.09±0.4）%、（0.72±0.11）%；Th17比例在HBV-SACLF组与CHB组及对照组比较，均明显升高，差异具有显著性（P_2=0.003，P30.001）。 2.HBV-SACLF早期、中期、晚期Th1、Th2、Th17的表达水平分别为：①Th1：（26.11±5.19）%、（22.45±4.06）%、（18.33±3.21）%；Th1比例在HBV-SACLF早期与中期，中期与晚期相比逐渐降低，两组之间比较差异均具有显著性(P1=0.001，P_2=0.004)。②T h2：（0.95±0.20）%、（1.66±0.41）%、（2.54±1.03）%；Th2比例在HBV-SACLF早期与中期，中期与晚期相比逐渐升高，两组之间比较差异均具有显著性(P1=0.003，P_2=0.048)。③Th17：（2.71±0.34）%、（3.39±0.73）%、（4.45±1.22）%；Th17比例在HBV-SACLF早期与中期，中期与晚期相比逐渐升高，两组之间比较差异均具有显著性(P1=0.002，P_2=0.007)。 3.HBV-SACLF感染组与非感染组Th1、Th2、Th17的表达水平分别为：①Th1：（23.59±3.91）%、（21.44±3.51）%；两组比较差异无显著意义。②T h2：（1.92±0.70）%、（1.76±0.55）%；两组比较差异无显著意义。③T h17：（4.65±1.33）%、（3.92±1.27）%，两组比较差异具有显著性（P=0.032）。 4.HBV-SACLF存活组与非存活组Th1、Th2、Th17的表达水平分别为：①Th1：（20.78±3.37）%、（25.61±4.05）%；两组比较差异无显著意义。②T h2：（1.65±0.59）%、（1.88±0.63）%；两组比较差异无显著意义。③T h17：（3.21±1.02）%、（4.53±1.29）%，两组比较差异具有显著性（P=0.029）。 5.对HBV-SACLF患者Th17、Th1、Th2与ALT进行Pearson相关性分析，结果发现Th17与ALT呈正相关（r=0.616，P=0.044），差异具有统计学意义；Th1、Th2与ALT无明显相关性。 6. HBV-SACLF组、肝衰竭前期组、CHB组、对照组IL-32、IL-10的表达水平分别为：①IL-32：（500.98±152.33）pg/ml、（486.45±129.06）pg/ml、（281.72±99.28）pg/ml、（178.16±50.54）pg/ml；IL-32在HBV-SACLF组与CHB组及对照组比较，均显著升高，差异具有显著性（P_2=0.021，P3=0.033）。②I L-10：（2.82±1.03）pg/ml、（3.05±1.83）pg/ml、（13.15±3.37）pg/ml、（7.62±2.17）pg/ml；IL-10在HBV-SACLF组与CHB组及对照组比较，均显著下降，差异具有显著性（P_2=0.024，P3=0.019）。 7. HBV-SACLF早期、中期、晚期IL-32、IL-10的表达水平分别为：①IL-32：（540.69±155.71）pg/ml、（498.43±135.56）pg/ml、（450.77±102.33）pg/ml；IL-32在HBV-SACLF早期与中期，中期与晚期相比逐渐降低，两组之间比较差异均具有显著性(P1=0.046，P_2=0.001)。②I L-10：（1.94±0.44）pg/ml、（2.83±0.97）pg/ml、（3.69±1.23）pg/ml；IL-10在HBV-SACLF早期与中期，中期与晚期相比逐渐升高，两组之间比较差异均具有显著性(P1=0.004，P_2=0.032)。结论Th1细胞和IL-32在HBV-SACLF患者中明显升高，随着病情发展呈逐渐下降趋势；Th2细胞和IL-10在HBV-SACLF患者中明显降低，其在HBV-SACLF早、中、晚期逐渐升高；Th17细胞在HBV-SACLF患者中明显升高，随着病情发展呈逐渐升高趋势；继发感染组Th17细胞较非感染组明显升高；存活组Th17细胞低于非存活组；且Th17细胞与肝细胞损害呈正相关。本文研究辅助性T细胞1、2、17及细胞因子IL-32、IL-10的紊乱是导致HBV-SACLF发生、发展的关键因素，通过检测其在肝衰竭早、中、晚期的动态变化可以判断机体的免疫状态，为临床的免疫调控治疗提供理论依据。
[Abstract]:Background chronic hepatitis B (CHB) is the main cause of chronic acute hepatic failure (SACLF) in China. It is reported that 80%., which accounts for all the causes of SACLF, is currently considered to be affected by many factors in the progression and prognosis of HBV infected patients, in which the immune function of the host plays an important role and the cytokine is regulating the immune response. It plays a key role. Auxiliary T cells (Th cells) are divided into Th0, Th1, Th2, and Th3 according to the secreted cytokines. At present, the most research is the two subtypes of Th1 and Th2. It is believed that in viral hepatitis infection, Th1 cells are beneficial to the clearance of viruses, while Th2 cells are mainly related to tissue damage and chronicity. The emergence of a new member of the cell family, -Th17 cell subgroup, rewrites the classic Th1/Th2 cell response pattern. This cell is characterized by hypersecreting IL-17 and has a strong pro-inflammatory role in.2005, Kim and others found IL-32, which can be produced by T cells, NK cells, monocytes, epithelial cells, and further studies show that they have a pro-inflammatory factor. Characteristic.IL-10 is a powerful immune and inflammatory inhibitor in the body, which is produced by Th2 cells, macrophages, dendritic cells (Dendritic cells, DC) and B cells, which can reduce the activity of T cells and reduce the effects of the immune response, resulting in the weakening of the body's ability to clear the virus. The changes of these cells and cytokines in CHB have already existed. It is reported whether it is involved in the process of HBV related chronic acute hepatic failure (HBV-SACLF) and the trend of dynamic changes in the course of liver failure, and the correlation with ALT has not been reported.
Objective to investigate the dynamic changes of Th1, Th2, Th17 and serum IL-32 and IL-10 in patients with HBV-SACLF and their clinical significance.
Methods the flow cytometry was used to detect 38 HBV-SACLF patients, 46 patients with pre liver failure, 20 CHB patients and 20 healthy persons (control group), the expression levels of Th1, Th2 and Th17 in the peripheral blood samples were compared and the differences between the above indexes were compared between the four groups; the trends of the early, middle and late Th1, Th2, Th17 in the HBV-SACLF patients were observed and compared. The changes in the expression level of Th1, Th2 and Th17 in the infected and non infected groups, the survival group and the non survival group, and the correlation between the Th1, Th2, Th17 and ALT were analyzed. At the same time, ELISA was used to detect the expression level of IL-32 and IL-10 in the four groups of serum, and IL-32, IL-10 was in the early, middle, and late stages of dynamic changes.
Results the expression levels of Th1, Th2, Th17 in the 1.HBV-SACLF group, the pre liver failure group, the CHB group and the control group were as follows: T h1: (22.58 + 3.86)%, (20.80 + 3.32)%, (5.23 + 1.51)%, (6.45 + 1.27)%, and Th1 ratio in both HBV-SACLF group and CHB group and control group, with significant difference (P_2=0.04, P3=0.032). (1.73 + 0.64)%, 1 (1.73 + 0.64)%), (1.73 + 0.64)%, (1)%, (1.73 + 0.64)%, (1.73 + 0.64)%), (1.73 + 0.64)%, (1)%, (1.73 + 0.64)%, (1.73 + 0.64)%, (1.73 + 0.64)%) .03 + 0.34)%, (6.15 + 0.97)%, (1.98 + 0.46)%, the proportion of Th2 in HBV-SACLF group and CHB group and control group decreased obviously, the difference was significant (P_2=0.007, P3=0.041), T h17: (3.48 + 0.80)%, (3.61 + 1.20)%, (1.09 + 0.4)%, (0.72 + 0.11)%, Th17 ratio was significantly higher in HBV-SACLF group than CHB group and control group, the difference has obviously increased, the difference has obvious It is significant (P_2=0.003, P30.001).
The expression level of 2.HBV-SACLF early, middle, late Th1, Th2, Th17 were respectively Th1: (26.11 + 5.19)%, (22.45 + 4.06)%, (18.33 + 3.21)%, Th1 ratio decreased gradually in the early and middle stages of HBV-SACLF, and the difference between the two groups was significant (P1=0.001, P_2=0.004). (2) T h2: (0.95 + 0.20)%, (1.66 + 0.41)%, 2.54 + 1.03)%; the ratio of Th2 was higher in the early and middle stages of HBV-SACLF than in the middle and late stages. The difference between the two groups was significant (P1=0.003, P_2=0.048). (2.71 + 0.34)%, (3.39 + 0.73)%, (4.45 + 1.22)%, and the proportion of Th17 was higher in the early and middle period of HBV-SACLF than in the middle and late stage, and the difference was compared between the two groups. They were both significant (P1=0.002, P_2=0.007).
The expression level of Th1, Th2, Th17 in 3.HBV-SACLF infection group and non infection group were as follows: (23.59 + 3.91)%, (21.44 + 3.51)%, and there was no significant difference in the two groups. (2) T h2: (1.92 + 0.70)%, (1.76 + 0.55)%, and there was no significant difference in two groups. (P=0) h17: (4.65 + 1.33)%, (3.92 +%)%, and there was significant difference (P=0, P=0). .032).
The expression levels of Th1, Th2, and Th17 in the 4.HBV-SACLF and non survival groups were as follows: (20.78 + 3.37)% and (25.61 + 4.05)%, and there was no significant difference in the two groups. (2) T h2: (1.65 + 0.59)%, (1.88 + 0.63)%, and there was no significant difference in two groups. (P=0) h17: (3.21 + 1.02)%, (4.53 +%)%, and there was significant difference (P=0, P=0). .029).
5. HBV-SACLF patients Th17, Th1, Th2 and ALT were analyzed with Pearson correlation. The results showed that Th17 was positively correlated with ALT (r=0.616, P=0.044), and the difference was statistically significant. Th1, there was no significant correlation between Th2 and ALT.
6. HBV-SACLF group, the expression level of IL-32, IL-10 in the pre liver failure group, CHB group and the control group were respectively IL-32: (500.98 + 152.33) pg/ml, (486.45 + 129.06) pg/ml, (281.72 + 99.28) pg/ml and (178.16 + 50.54) pg/ml, and IL-32 was significantly higher in HBV-SACLF group than in CHB and control groups. L-10: (2.82 + 1.03) pg/ml, (3.05 + 1.83) pg/ml, (13.15 + 3.37) pg/ml, (7.62 + 2.17) pg/ml, and IL-10 in HBV-SACLF group were significantly lower than those of CHB and control groups, and the difference was significant (P_2=0.024, P3=0.019).
7. HBV-SACLF early, middle and late IL-32, IL-10 expression levels are: IL-32: (540.69 + 155.71) pg/ml, (498.43 + 135.56) pg/ml, (450.77 + 102.33) pg/ml; IL-32 in the early and middle stages of HBV-SACLF, compared to the middle and late gradually decreased, between the two groups has a significant difference (P1=0.046, P_2=0.001). (1.94 + 0 I). .44) pg/ml, (2.83 + 0.97) pg/ml, (3.69 + 1.23) pg/ml; IL-10 increased gradually in the early and middle stages of HBV-SACLF, compared to the middle and late stages, and the difference was significant between the two groups (P1=0.004, P_2=0.032).
Conclusion Th1 cells and IL-32 were significantly increased in HBV-SACLF patients, and gradually decreased with the development of the disease. Th2 cells and IL-10 were obviously decreased in HBV-SACLF patients. It was gradually increased in the early, middle and late stages of HBV-SACLF, and Th17 cells increased significantly in HBV-SACLF patients, and gradually increased with the development of the disease; secondary infection group Th17. The cells in the survival group were significantly lower than those in the non infected group; the Th17 cells in the survival group were lower than those in the non surviving group; and the Th17 cells were positively related to the liver cell damage. This paper studied the auxiliary T cell 1,2,17 and the cytokine IL-32, and the disorder of IL-10 was the key factor leading to the occurrence of HBV-SACLF, and the dynamic changes in the early, middle and late stages of the liver failure could be detected. To determine the immune state of the body, and to provide a theoretical basis for clinical immune regulation and treatment.

【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R575.3;R512.62

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