格林—巴利综合征患者血清IL-22及IL-37水平的变化及其临床意义

发布时间：2018-05-08 23:00

本文选题：格林-巴利综合征 + 白细胞介素-22　；参考：《郑州大学》2014年硕士论文

【摘要】：研究目的与背景格林—巴利综合征(Guillain—Barre Syndrome,GBS)是一种常见的多发性脊神经和周围神经的脱髓鞘疾病，由体液免疫及细胞免疫共同介导，又称为对称性多神经根炎或急性特发性多神经炎。临床上表现为进行性上升性对称性麻痹、四肢软瘫，以及不同程度的感觉障碍。细胞因子在自身免疫损伤中起到启动、维持和调节的作用，细胞因子在GBS及其动物模型ENA中的作用受到越来越多的学者关注，经大量实验研究表明，免疫性疾病的发展与转归是由促炎性细胞因子及抑炎性细胞因子介导的Th1/Th2型细胞间平衡的改变造成的。白细胞介素-22(Interleukin-22,IL-22)是最近发现的IL-10细胞因子家族中的成员之一，NK细胞、γδT细胞、LTi细胞、Th17细胞、Th22细胞均可产生IL-22，其中Th17细胞是一种CD4+T效应细胞，可分泌多种细胞因子，如IL-2、IL-17A、IL-17F、IL-22，在实验性自身免疫性脑炎的小鼠模型（EAE）中，IL-17mRNA基因水平和蛋白质水平均升高，IL-22及IL-17在多种体外试验及动物模型中被证实可相互协同，共同参与促炎性免疫反应，同时，在肿瘤细胞中，IL-22可诱导结合珠蛋白、血清淀粉样蛋白、α1抗胰凝乳蛋白酶等急性反应期蛋白的产生。白介素-37（Interleukin-37,IL-37）是IL-1细胞因子家族中的成员之一，IL-37在巨噬细胞或上皮细胞内表达，可以抑制TLR诱导的促炎因子IL-1α、IL-1β、IL-18、IFN-γ和TNF-α的产生、释放以及促进树突状细胞的活性，也可以促进CD4+CD25+调节性T细胞的表达，从而有效的抑制免疫应答。此次研究通过检测GBS患者急性期及恢复期血清中IL-22及IL-37的浓度，探讨这两种细胞因子在GBS发病及转归的过程中可能起到的作用，为新的GBS治疗方案提供进一步的理论基础。资料和方法收集2012年9月至2014年2月间在郑州大学第一附属医院神经内科住院的GBS患者共51例，其中男性患者30例，女性患者21例，年龄8~74岁，平均(42.00±18.23)岁。所有入选的GBS患者均经过我院神经内科两名以上医师详细询问病史、神经系统专科体检、肌电图检查及脑脊液检测，检查结果符合Asbury等于1990年制定的诊断标准，然后对所有入选患者进行Hughes评分来评测疾病的严重程度。选取同一时期在我院门诊体检的健康志愿(Healthy Control，HC)者31例为对照组，其中男性19例，女性12例，年龄8~66岁，平均(40.61±18.23)岁。抽血前无手术史、免疫治疗史，无感染性疾病、肿瘤、心脑血管疾病等。实验组和对照组年龄及性别差别无统计学意义。在采血前，每位健康人及患者需禁食、禁水10小时以上。在第二日清晨抽取肘静脉血3ml，随后置于真空采血管中（无抗凝剂），，放入-80℃冰箱备用。GBS患者组均于急性期（起病l～14天）和恢复期（起病30～60天）分别采集肘静脉血标本。健康对照组仅采集一次静脉血标本。采用双抗体夹心酶联免疫吸附方法(Enzyme Linked Immunosorbent Assay,ELISA)测定血清中IL-22和IL-37的水平。采用SPSSl7．0软件包处理所有数据，应用均数±标准差(x±s)表示定量资料。运用两独立样本t检验进行两组定量资料的比较，运用Spearman相关系数分析两组定量资料的相关性，运用单因素方差分析进行多组定量资料的比较，随后采用组间比较(LSD检验)法，若P0.05，表示差异有统计学意义。结果 1.GBS患者组急性期、恢复期与对照组血清IL-22水平的比较：GBS患者组急性期血清IL-22水平(110.70±24.37)ng/L较恢复期(78.32±11.68)ng/L显著升高，P=0.001；较对照组(75.62±15.00)ng/L显著升高，P=0.000；GBS患者组恢复期血清IL-22水平虽有所下降，但仍较对照组升高，P=0.030。 2.GBS患者组恢复期、急性期与对照组血清IL-37水平的比较：GBS患者组恢复期血清IL-37水平(84.38±6.67)ng/L较急性期(62.80±9.29)ng/L显著升高，P=0.004；较对照组(57.82±16.11)ng/L显著升高，P=0.000；GBS患者组急性期血清IL-37水平虽较恢复期降低，但仍较对照组升高，P=0.039。 3.GBS患者急性期血清IL-22及IL-37水平与疾病严重程度的关系：GBS患者急性期血清中IL-22水平与Hughes评分显著相关，r=0.803，P=0.001；GBS患者急性期血清中IL-37水平与Hughes评分无明显相关性，r=-0.246，P=0.082。 4.GBS患者恢复期血清IL-22及IL-37水平与疾病严重程度的关系：GBS患者恢复期血清中IL-22水平与Hughes评分无明显相关性，r=0.029，P=0.839；GBS患者恢复期血清中IL-37水平与Hughes评分显著相关，r=-0.766，P=0.000。 5.各类型GBS患者急性期血清IL-22及IL-37水平的比较：脱髓鞘+轴索型GBS患者急性期血清IL-22水平(141.13±23.45)ng/L较轴索型(108.41±9.25)ng/L升高，P=0.002，较脱髓鞘型(90.45±3.62)ng/L显著升高，P=0.000,轴索型GBS患者急性期血清IL-22水平较脱髓鞘型升高,P=0.004；脱髓鞘+轴索型、轴索型、脱髓鞘型患者急性期血清中IL-37水平的差异无统计学意义，F=3.098P=0.054。 6.各类型GBS患者恢复期血清IL-22及IL-37水平的比较：脱髓鞘+轴索型、轴索型、脱髓鞘型患者恢复期血清中IL-22水平的差异无统计学意义，F=0.473P=0.626；脱髓鞘型GBS患者恢复期血清IL-37水平(90.10±3.72)ng/L，较轴索型(84.44±3.06)ng/L升高，P=0.010，较脱髓鞘+轴索型(75.52±3.30)ng/L显著升高，P=0.000轴索型GBS患者恢复期血清IL-37水平，较脱髓鞘+轴索型升高，P=0.005。结论 1.IL-22在GBS患者急性期与恢复期血清中的表达水平均较对照组显著升高，推测IL-22可能参与GBS免疫应答。GBS患者急性期IL-22水平较恢复期升高更显著，考虑IL-22可能主要在免疫应答起始阶段起促炎作用。 2.IL-37在GBS患者急性期与恢复期血清中的表达水平均较对照组显著升高，考虑IL-37可能参与GBS免疫应答。GBS患者恢复期IL-37水平较急性期升高更显著，考虑IL-37可能主要在疾病恢复期起抑炎性作用。 3.IL-22在GBS患者急性期血清中的表达水平与疾病严重程度呈正相关，IL-22水平越高，病情越重；IL-37在GBS患者恢复期血清中的表达水平与疾病严重程度呈负相关，IL-37水平越高，病情越轻。 4.GBS患者急性期IL-22升高可能参与髓鞘脱失及轴索的损害，其显著升高可能提示预后不良，GBS患者恢复期IL-37升高可能参与髓鞘及轴索的修复，其显著升高可能提示预后较好。
[Abstract]:Research purpose and background
Green Barre syndrome (Guillain Barre Syndrome, GBS) is a common multiple spinal and peripheral nerve demyelinating disease, which is mediated by humoral immunity and cell immunity. It is also known as symmetric multiple neuropathy or acute idiopathic multiple neuritis. It is clinically manifested as progressive symmetrical paralysis and soft paralysis of the limbs. As well as different degree of sensory disturbance. Cytokine plays the role of initiation, maintenance and regulation of autoimmune injury. The role of cytokines in GBS and its animal model ENA is concerned by more and more scholars. A large number of experimental studies have shown that the development and outcome of immune diseases are derived from proinflammatory cytokines and anti inflammatory cells. Factor mediated changes in the balance between Th1/Th2 type cells. Interleukin-22 (IL-22) is one of the members of the recently discovered IL-10 cell factor family, NK cells, NK cells, LTi cells, Th17 cells, Th22 cells can produce IL-22, and Th17 cells are a kind of effector cells, which can secrete a variety of cytokines. Such as IL-2, IL-17A, IL-17F, IL-22, in the mouse model of experimental autoimmune encephalitis (EAE), the IL-17mRNA gene level and protein level are increased. IL-22 and IL-17 have been confirmed in a variety of in vitro experiments and animal models to cooperate with each other to participate in the inflammatory response, and in the tumor cells, IL-22 can induce the combination of Pearl eggs. White, serum amyloid, alpha 1 anti - pancreatic chymotrypsin and other acute reaction proteins. -37 (Interleukin-37, IL-37) is one of the members of the IL-1 cell factor family. IL-37 is expressed in macrophages or epithelial cells, which inhibits the production of TLR induced proinflammatory cytokines, IL-1 a, IL-1 beta, IL-18, IFN- gamma, and TNF- alpha. And promoting the activity of dendritic cells can also promote the expression of CD4+CD25+ regulatory T cells, and thus effectively inhibit the immune response. This study explored the possible role of these two cytokines in the pathogenesis and return of GBS by detecting the concentrations of IL-22 and IL-37 in the serum of GBS patients at acute and convalescent stages, as a new GBS. The treatment scheme provides a further theoretical basis.
Information and methods
A total of 51 GBS patients were hospitalized in the Department of Neurology of the First Affiliated Hospital of Zhengzhou University from September 2012 to February 2014. There were 30 male patients, 21 female patients, age 8~74 years, average (42 + 18.23) years old. All the selected GBS patients were examined by more than two physicians in the Department of Neurology in our hospital. The results of electromyography and cerebrospinal fluid examination were in accordance with the diagnostic criteria of Asbury equal to 1990, and then Hughes scores were used to evaluate the severity of the disease. 31 cases of Healthy Control (HC) in the same period of our hospital were selected as the control group, including 19 males and 12 females. The average age of 8~66 years was (40.61 + 18.23) years old. There was no history of operation, history of immunotherapy, no infectious disease, tumor, cardiovascular and cerebrovascular diseases before blood extraction. There was no statistical difference between the age and sex of the experimental group and the control group.
Before the blood collection, every healthy person and patient need to fasting and prohibit water for more than 10 hours. Extraction of 3ml in the elbow vein blood on the early morning of second, then placed in the vacuum blood vessel (no anticoagulant), and put into the acute phase (L to 14 days) and the recovery period (30~60 days of the onset of disease) to collect the blood specimens of the elbow vein, and the healthy control group. Only one venous blood specimen was collected. The level of IL-22 and IL-37 in serum was measured by Enzyme Linked Immunosorbent Assay (ELISA). All data were processed with SPSSl7.0 software package, and the quantitative data were expressed by mean standard deviation (x + s). Two groups of quantitative data were carried out by two independent sample t test. In comparison, the correlation of two groups of quantitative data was analyzed with Spearman correlation coefficient, and the comparison of multiple quantitative data was carried out by single factor analysis of variance. Then group comparison (LSD test) method was used, if P0.05, the difference was statistically significant.
Result
In the acute phase of 1.GBS patients, the comparison of serum IL-22 levels in the acute phase and the control group: the level of serum IL-22 in the acute phase of the GBS patients (110.70 + 24.37) ng/L was significantly higher than that in the recovery period (78.32 + 11.68) ng/L, P=0.001; the ng/L was significantly higher than the control group (75.62 + 15) ng/L, P=0.000, although the serum IL-22 level in the recovery period of the GBS patients decreased, but still remained. Higher than the control group, P=0.030.
In the convalescent period of 2.GBS patients, the comparison of serum IL-37 levels between the acute phase and the control group: the level of serum IL-37 in the convalescent period of the GBS patients (84.38 + 6.67) was significantly higher than that in the acute phase (62.80 + 9.29) ng/L, P=0.004; the ng/L was significantly higher than the control group (57.82 + 16.11) ng/L, and the serum IL-37 level in the acute phase of the GBS patients was lower than the recovery period, but the level of serum IL-37 was lower than that of the control group, but the serum IL-37 level was lower than the recovery period. It is still higher than the control group, P=0.039.
The relationship between serum IL-22 and IL-37 levels in the acute phase of 3.GBS and the severity of the disease: the level of IL-22 in serum of patients with GBS was significantly correlated with the Hughes score, r=0.803, P=0.001, and there was no significant correlation between the IL-37 level and Hughes score in the acute phase of GBS patients.
The relationship between the level of serum IL-22 and IL-37 and the severity of the disease in the convalescent period of 4.GBS patients: there was no significant correlation between the serum IL-22 level and the Hughes score in the recovery period of GBS patients, r=0.029, P=0.839, and the IL-37 level in the serum of GBS patients was significantly related to the Hughes score. R=-0.766
5. comparison of serum IL-22 and IL-37 levels in acute phase of GBS patients: the level of serum IL-22 in patients with demyelinating + axonal GBS (141.13 + 23.45) ng/L was higher than that of axonal type (108.41 + 9.25) ng/L, P=0.002, and demyelinating type (90.45 + 3.62) ng/L increased significantly. P=0.000, the serum IL-22 level of axonal GBS patients was higher than demyelinating type. There was no significant difference in serum IL-37 level between the high and P=0.004, demyelination + axonal type, axonal type and demyelinating patients, F=3.098P=0.054.
6. the comparison of serum IL-22 and IL-37 levels in the recovery period of all types of GBS patients: the difference in serum IL-22 levels in demyelinating + axonal type, axonal type and demyelinating patients was not statistically significant, F=0.473P=0.626; the serum IL-37 level (90.10 + 3.72) ng/L in demyelinating GBS patients was higher than that of axonal type (84.44 + 3.06) ng/L, P=0.010, Compared with demyelination + axonal type (75.52 + 3.30) ng/L, the serum IL-37 level of P=0.000 axonal GBS patients was higher than that of demyelination + axonal type, P=0.005.
conclusion
The expression level of 1.IL-22 in the acute and convalescent serum of GBS patients was significantly higher than that in the control group. It is suggested that IL-22 may participate in the acute phase of GBS immune response to.GBS patients with higher level of IL-22 than that of the recovery period, and IL-22 may be used primarily in the initial stage of the immune response.
The expression level of 2.IL-37 in the acute and convalescent serum of GBS patients was significantly higher than that in the control group. Considering that IL-37 may participate in the GBS immune response of.GBS patients, the level of IL-37 was more significant in the convalescence period, and IL-37 might be mainly in the recovery period of the disease.
The expression level of 3.IL-22 in the acute phase of GBS patients was positively correlated with the severity of the disease, the higher the level of IL-22, the heavier the disease, and the negative correlation between the serum level of IL-37 in the recovery period of GBS patients and the severity of the disease, the higher the level of IL-37, the lighter the disease was.
The elevation of IL-22 in patients with 4.GBS may be involved in myelin loss and axonal damage. The significant increase may indicate poor prognosis. The increase of IL-37 in the recovery period of GBS may be involved in the repair of myelin and axon, which may suggest a better prognosis.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R744

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