IL-6对视神经脊髓炎谱系病患者Th17/Treg失衡的调控机制研究

目的 研究调控白细胞介素(IL)-6对视神经脊髓炎谱系病(NMOSD)患者辅助性17型T细胞(Th17)/调节性T细胞(Treg)比值失衡的影响。方法 收集2019年1月至2020年12月就诊的9例AQP4-IgG阳性的NMOSD患者为NMOSD组和同期8例健康志愿者为对照组,收集并培养外周血单个核细胞(PBMC)后,分别予以IL-6或IL-6受体抑制剂共培养,采用流式细胞术检测Th17和Treg比例,流式微球检测技术检测调控IL-6水平后对Th17、Treg、IL-17、IL-10水平的影响。结果 NMOSD组PBMC细胞添加IL-6共培养后,可见Th17比例及Th17/Treg比值增高(均P<0.05)。添加IL-6阻滞剂共培养后,IL-10水平下降,Treg表达增加,Th17/Treg比值下降(均P <0.05)。结论 NMOSD患者PBMC中的外源性IL-6可以上调Th17比例而导致Th17/Treg比值失衡;但外源性IL-6受体阻滞剂可以升高IL-10水平,通过增加Treg比例恢复Th17/Treg比值平衡。     

曲美他嗪对急性脑梗死患者的临床疗效及相关机制探讨

目的 探讨曲美他嗪辅助治疗急性脑梗死(acute cerebral infarction,ACI)患者的临床疗效、不良反应及对Th17/Treg细胞及其相关细胞因子水平的影响。方法 纳入南阳市中心医院84例急性脑梗死患者为研究对象,分为对照组(n=42)和观察组(n=42)。对照组采用常规治疗,观察组在常规治疗的基础辅助曲美他嗪口服(3次/d,20mg/次),2组均治疗2周。观察2组患者治疗后的临床疗效、不良反应及Th17比例、Treg比例、Th17/Treg比值及其相关细胞因子水平。结果 2周后,观察组和对照组总有效率分别为97.62%和83.33%,与对照组相比,观察组总有效率显著升高(97.62%vs 83.33%,P=0.026)。2组患者治疗前Th17比例、Treg比例、Th17/Treg比值、血清IL-17A和TGF-β水平相比,差异均无统计学意义(P0.05);2周后,与对照组相比,观察组Th17比例、Th17/Treg比值和血清IL-17A水平显著降低,而Treg比例及血清TGF-β水平显著升高,差异均有统计学意义(P0.05)。2组不良反应发生率比较差异无统计学意义(P=0.724)。结论 曲美他嗪辅助治疗ACI临床疗效显著,可能与改善ACI患者Th17/Treg细胞平衡有关,且不增加药物不良反应。     

帕金森病患者外周血Th17/Treg平衡及相关细胞因子水平测定及意义

目的探讨帕金森病(PD)患者外周血Th17/Treg平衡及相关细胞因子含量变化的意义。方法收集50例PD患者(PD组)及50例健康体检者(Control组),采集所有研究对象外周血。采用流式细胞仪检测外周血中Th17细胞及Treg细胞含量;采用ELISA检测血浆中细胞因子TGF-β、IL-6和IL-17水平。结果流式细胞计数结果显示,PD组的外周血Treg细胞和Th17细胞水平较Control组均显著增加(P均0.01),Treg细胞的增长程度明显大于大于Th17细胞,同时PD组Treg/Th17细胞比较Control组也明显增加(P均0.01)。ELISA检测结果显示,与Control组相比,PD组的血浆TGF-β1、IL-6及IL-17含量均有显著增加(P均0.01)。结论 PD患者的外周血Treg/Th17平衡存在失衡,提示PD病理过程可能与其外周血Treg/Th17失衡有关。     

急性脑梗死患者外周血Th17、Treg、Th17/Treg及炎症因子的动态变化过程研究

目的探讨急性脑梗死(AIS)患者外周血Th17、Treg、Th17/Treg及相关炎症因子的表达及其动态演变过程。方法选取32例AIS患者为实验组及32名健康人为对照组。分别检测AIS患者第1 d、4 d、7 d,外周血中Th17及Treg占CD4+T细胞比例及Th17/Treg比值,并检测白介素6(IL-6)、中性粒细胞与淋巴细胞比值(NLR)、C反应蛋白(CRP)和红细胞沉降率(ESR),分析其在AIS后的变化规律。结果AIS组于起病后第1 d、4 d,外周血Th17、Treg比例分别与对照组相比,差异有统计学意义(均P<0.05);AIS患者第7 d Th17、Treg比例逐渐恢复,与对照组比较差异无统计学意义(均P>0.05);AIS患者Th17/Treg比值在第1 d、4 d、7 d逐渐减小,但始终明显高于对照组(均P<0.05)。在AIS后的第1 d、4 d、7 d,IL-6、NLR及CRP分别均明显高于对照组(均P<0.05);AIS患者ESR第1 d与对照组比较无统计学意义(P>0.05),第4 d、7 d明显高于对照组(均P<0.05)。AIS患者外周血Th17比例及Th17/Treg、NLR、CRP、ESR分别与IL-6呈正相关(ρ=0.298,P=0.007;ρ=0.351,P=0.001;ρ=0.377,P=0.001;ρ=0.582,P=0.000;ρ=0.388,P=0.000);Treg比例与IL-6呈负相关(ρ=-0.242,P=0.029)。结论AIS患者早期存在Th17/Treg失衡,且合并有全身炎症反应。     

Th17/Treg细胞及相关细胞因子在复发缓解型多发性硬化中的作用及机制

目的探讨辅助性T细胞(Th17)、调节性T细胞(Treg)和相关细胞因子在复发缓解型多发性硬化(RRMS)患者发病机制中的作用与机制。方法收集急性期RRMS患者(RRMS组)31例,以神经系统非炎性疾病29例为对照组。RRMS患者入院后给予静脉滴注甲泼尼龙1000 mg/d冲击治疗,后续每3 d剂量减半。对RRMS组患者治疗前和甲泼尼龙治疗2周后(治疗后)分别进行残疾状况拓展性量表(EDSS)评分;采用流式细胞术(FCM)检测RRMS组患者治疗前后及对照组患者外周血Th17(CD4~+IL-17~+)细胞和Treg(CD4~+FOXP3~+)细胞百分率;采用酶联免疫吸附试验(ELISA)检测RRMS组患者治疗前后及对照组患者外周血血浆中白细胞介素(IL)-17A、IL-23、IL-6、IL-10、IL-35和转化生长因子-β(TGF-β)水平;采用Spearman相关分析对RRMS组患者治疗前外周血Th17细胞数量与IL-17A、IL-23、IL-35、IL-6、IL-10及TGF-β水平进行相关性分析。结果 (1)RRMS组治疗前EDSS评分高于治疗后[分别(6.31±1.54)分vs.(4.02±0.68)分,t=0.75,P0.05];(2)FCM分析结果显示,与对照组[(3.12±1.27)%]比较,RRMS组患者治疗前Th17细胞百分率[(15.24±2.54)%]明显升高(P0.05),而对照组Treg细胞百分率[(35.04±4.21)%]明显高于RRMS组治疗前[(11.12±3.13)%,P0.05];与对照组(0.10±0.02)相比,RRMS组治疗前Th17/Treg(1.51±0.62)也明显升高(P0.01);与RRMS组治疗前[(11.12±3.13)%]比较,甲泼尼龙治疗后Treg细胞比例[(23.14±2.86)%]明显升高(P0.01);与RRMS组治疗前Th17细胞百分率[(15.24±2.54)%]相比,甲泼尼龙治疗后Th17细胞百分率[(4.24±1.14)%]明显降低(t=0.88,P0.05);与RRMS组治疗前(1.51±0.62)相比,甲泼尼龙治疗后Th17/Treg比值(0.19±0.07)降低(t=0.95,P0.01);(3)ELISA法检测结果显示,RRMS组治疗前IL-17A[分别(17.26±1.21)pg/mL vs.(3.23±0.81)pg/mL,t=0.72,P0.05]、IL-23[(分别(64.38±7.51)pg/mL vs.(21.14±1.82)pg/mL,t=0.75,P0.05]、IL-6[分别(70.14±8.17)pg/mL vs.(7.28±0.75)pg/mL,t=0.95,P0.01]和IL-10水平[分别(21.12±2.74)pg/mL vs(2.39±0.34)pg/mL,t=0.91,P0.01]均明显高于对照组,而IL-35[分别(0.31±0.06)pg/mL vs.(1.55±0.16)pg/mL,t=-0.89,P0.01]和TGF-β水平[分别(5.13±0.34)pg vs.(18.25±0.74)pg/mL,t=-0.83,P0.05]显著低于对照组;与RRMS组治疗前比较,治疗后IL-17A[分别(17.26±1.21)pg/mL vs.(4.23±0.90)pg/mL,t=0.82,P0.05]、IL-23[分别(64.38±7.51)pg/mL vs.(29.73±2.51)pg/mL,t=0.77,P0.05]和IL-6[分别(70.14±8.17)vs.(15.23±1.86),t=0.89,P0.01]水平明显降低,IL-35(分别(0.31±0.06)pg vs.(1.41±0.18)pg,t=-0.88,P0.01)和TGF-β水平[分别(5.13±0.34)pg vs.(16.91±0.59)pg,t=-0.72,P0.05]明显升高;(4)RRMS组患者Th17细胞与IL-17A(r=0.89,P0.01)、IL-23(r=0.71,P0.05)和IL-6(r=0.83,P0.05)水平呈正相关,与IL-35和TGF-β水平呈负相关[分别r=-0.82,P0.01;r=-0.74,P0.05]。结论 RRMS的发生可能与Th17细胞上调,Treg细胞下调,Th17/Treg表达失衡,相关细胞因子IL-17A、 IL-23、IL-6水平升高,IL-35和TGF-β水平降低有着密切关系。     

Th17/Treg细胞在复发缓解型多发性硬化中的表达

目的探讨辅助T细胞17(Th17)和调节性T细胞(Treg)相关细胞因子白细胞介素17(IL-17)和白细胞介素35(IL-35)在复发缓解型多发性硬化(RRMS)发病机制中的作用及甲泼尼龙对其表达的影响。方法收集急性期RRMS患者36例,神经系统非炎性疾病患者32例作为对照组,采用酶联免疫法(ELISA)检测两组患者治疗前及治疗后2周时血清和脑脊液中IL-17、IL-35的水平。结果 RRMS组治疗前血清和脑脊液IL-17水平显著高于对照组(P0.01),而其IL-35水平均低于对照组(P0.05)。与治疗前相比,RRMS组治疗后血清和脑脊液IL-17下降(P0.01),而IL-35升高(P0.05)。RRMS组治疗前血清中IL-17水平与IL-35水平呈负相关(r=-0.551,P=0.041)。结论 Th17/Treg表达的失衡与RRMS的发病有关,甲泼尼龙治疗可使血清、脑脊液IL-17水平下降,IL-35水平升高,进而抑制炎性反应。     

Th17细胞及其效应分子IL-17在HAM/TSP患者体内的变化及Tax蛋白对IL-17的影响

目的 研究Th17细胞及其效应分子IL-17在HTLV-Ⅰ相关性脊髓病/热带痉挛性截瘫(HTLV-Ⅰassociated myelopsthy/tropical spastic paraparesis,HAM/TSP)患者中的变化及Tax蛋白对IL-17的影响.方法 分别应用ELISA法和流式细胞技术检测HAM/TSP患者CSF上清液中的IL-17水平及外周血Th17细胞的百分率.应用RNAi干扰技术抑制Tax蛋白的表达,并用ELISA法检测干扰后细胞培养悬浮液中IL-17的表达.结果 HAM/TSP患者脑脊液上清中IL-17含量(4.58±0.70) pg/mL较对照组(0.76±0.17) pg/mL显著升高(P＜0.01);HAM/TSP患者外周血中Th17细胞的百分率(2.00％±0.64％)较对照组(0.41％±0.24％)显著升高(P＜0.01).RNAi技术干扰Tax蛋白表达后,IL-17水平(5.04± 1.27) pg/mL较未干扰组细胞培养悬浮液中IL-17水平(7.69±2.11) pg/mL显著降低(P＜0.05).结论 Th17细胞及其效应分子IL-17在HAM/TSP患者体内显著提高,提示在其发病机制中发挥一定作用,而且IL-17的表达水平受Tax蛋白调控.     

颅内破裂动脉瘤手术时机的临床研究

目的 探讨颅内破裂动脉瘤不同手术时机患者的血清炎性因子水平变化及其临床意义。方法 回顾性分析2017年6月-2019年12月在本院神经外科确诊的颅内动脉瘤性蛛网膜下腔出血患者180例,根据医师建议和患者自愿分为早期治疗组(入院1～3 d进行手术治疗)和延期治疗组(入院4～14 d进行手术治疗),比较2组患者手术前后血清炎性细胞因子水平变化,并对2组患者的疗效和术后并发症发生情况进行比较。结果 手术前2组血清TNF-α,IL-6,IL-1β水平无显著差异(P>0.05); 2组术后第8 d血清TNF-α,IL-6,IL-1β水平均较术前有所降低(P<0.05); 早期治疗组术后第8 d血清TNF-α,IL-6,IL-1β水平显著低于延期治疗组(P<0.05)。早期治疗组的总有效率显著高于延期治疗组(P<0.05); 早期治疗组的恢复良好率显著高于延期治疗组(P<0.05); 早期治疗组的致残率显著低于延期治疗组(P<0.05)。2组重残率、植物生存率及病死率无显著差异(P>0.05)。早期治疗组的颅内感染和脑血管痉挛的发生率明显低于延期治疗组(P<0.05),脑积水和再出血的发生率没有显著差异(P>0.05)。结论 早期治疗的颅内破裂动脉瘤患者的术后血清炎性指标水平变化较显著,疗效及预后均较好,并发症发生率低; 血清炎性指标水平可能与患者的疗效和预后有关,可作为aSAH发生发展的预测指标。     

颅内血肿微创清除术联合吡拉西坦治疗脑出血的疗效及对血清HMGB-1、IGF-1水平的影响

目的 观察颅内血肿微创清除术联合吡拉西坦治疗脑出血的疗效及对血清HMGB-1、IGF-1水平的影响。方法 收集2016年1月-2017年12月于本院神经外科住院的脑出血患者,按照数字随机分组法分为观察组和对照组,观察组采用颅内血肿微创清除术联合吡拉西坦,对照组采用颅内血肿微创清除术,比较2组患者的疗效、治疗前后血清HMGB-1、IGF-1水平的变化及不良反应。结果 观察组的有效率为95.0%,高于对照组(75.0%)(P<0.05); 治疗前2组患者的血清HMGB-1分别为(7.43±0.31)ng/mL、(7.36±0.35)ng/mL,治疗后分别为(1.54±0.46)ng/mL、(2.90±0.48)ng/mL,治疗后2组患者的血清HMGB-1水平均有所降低(P<0.05),且观察组较对照组下降明显(P<0.05); 治疗前2组患者的血清IGF-1水平分别为(31.21±5.12)、(31.18±4.95),治疗后分别为(52.33±6.24)、(45.48±5.91),治疗后2组患者的血清IGF-1水平均有所升高(P<0.05),且观察组较对照组升高明显(P<0.05); 2组患者不良反应发生率分别为20.0%、12.5%,2组比较无明显差异(P>0.05)。结论 颅内血肿微创清除术联合吡拉西坦治疗脑出血能够显著提高其治疗效果,有助于降低血清HMGB-1水平,提高血清IGF-1水平,且安全性较好。     

Tc1/Tc2 and Th1/Th2 balance in Asian and Western types of multiple sclerosis, HTLV-I-associated myelopathy/tropical spastic paraparesis and hyperIgEaemic myelitis

CD8+ T cells, like CD4+ T cells, can differentiate into at least two subsets with distinct cytokine patterns: Tc1 cells produce Th1-like cytokines and Tc2 cells produce Th2-like cytokines. To clarify the immunopathological roles of Tc1 and Tc2 cells in central nervous system (CNS) inflammation, we examined intracellular cytokines in CD8+ and CD4+ T cells by flow cytometry and analyzed the Tc1/Tc2 balance as well as the Th1/Th2 balance in 80 patients with various CNS inflammatory diseases, including 20 with optico-spinal multiple sclerosis (OS-MS), 21 with conventional MS (C-MS), 22 with human T-lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and 17 with hyperIgEaemic myelitis. Twenty-two healthy subjects were also examined as controls. Patients with OS-MS showed a significantly higher percentage of INF-gamma+IL-4- CD8+ T cells as well as CD4+ T cells and a significantly higher intracellular interferon-gamma (IFN-gamma)/interleukin-4 (IL-4) ratio both in CD8+ and CD4+ T cells throughout the relapse and remission phases than the healthy controls. Furthermore, the patients with OS-MS showed a significantly lower percentage of INF-gamma-IL-4+ CD4+ T cells as well as CD8+ T cells during the relapse phase than the healthy controls. On the other hand, the patients with C-MS showed a significantly higher percentage of IFN-gamma-IL-4+ CD8+ T cells in addition to more IFN-gamma+IL-4- CD4+ T cells during the relapse phase than the healthy controls. The HAM/TSP patients showed a significantly higher percentage of INF-gamma+IL-4- CD8+ T cells and a significantly higher intracellular IFN-gamma/IL-4 ratio in CD8+ T cells than the healthy controls. In contrast, in hyperIgEaemic myelitis, in addition to a significantly lower intracellular IFN-gamma/IL-4 ratio in CD4+ T cells, a tendency toward a lower intracellular IFN-gamma/IL-4 ratio in CD8+ T cells in comparison to the healthy controls was observed. These results clarified for the first time the distinct Tc1/Tc2 balance in each disease condition as follows: Tc1 cell response is predominant in OS-MS and HAM/TSP, while Tc2 cell response is predominant in hyperIgEaemic myelitis and at relapse phase of C-MS. Furthermore, our results suggest that CD8+ T cells play an adjunctive role in disease induction and the clinical course of MS.     

Long-term favorable response to interferon beta-1b is linked to cytokine deviation toward the Th2 and Tc2 sides in Japanese patients with multiple sclerosis

To address the immune mechanism of the long-term beneficial effects of interferon beta (IFN-beta), we measured the intracellular cytokine production patterns of IFN-gamma, IL-4 and IL-13 in peripheral blood CD4+ and CD8+ T cells, which previously displayed alterations during the early course of IFN-beta treatment, in 15 Japanese patients after long-term IFN-beta administration. The patients were treated with IFN-beta-1b 8 x 10(6) units given subcutaneously every other day for a mean period of 34.5 +/- 5.5 months (range: 26-43 months). During the follow-up period, 6 patients experienced 33 relapses, while the other 9 were relapse-free. The results revealed the following cytokine alterations: (1) type 2 cytokine, such as IL-4 and IL-13, were significantly increased in producing cell percentages in both CD4+ (p = 0.0356 and p = 0.0007, respectively) and CD8+ (p = 0.0231 and p = 0.0170, respectively) T cells while IFN-gamma, a representative type 1 cytokine, was significantly decreased in the absolute producing cell numbers (p = 0.0125 in CD4+ T cells and p = 0.0022 in CD8+ T cells) even after approximately 3 years of IFN-beta administration; (2) the intracellular IFN-gamma / IL-4 ratio tended to decrease in both CD4+ and CD8+ T cells (p = 0.0535 and p = 0.0783, respectively), reflecting a strong downmodulation of type 1 cytokine producing cells; and importantly (3) alterations such as the decreased intracellular IFN-gamma / IL-4 ratio in CD4+ T cells and increased percentage of CD8+ IL-13+ T cells compared with the pretreatment levels were only statistically significant in MS patients without relapse during IFN-beta therapy (p = 0.0152 and p = 0.0078, respectively). Therefore, we consider that cytokine deviation toward the Th2 and Tc2 sides is linked to a long-term favorable response to IFN-beta, while a higher intracellular IFN-gamma / IL-4 ratio is associated with treatment failure.     

α-硫辛酸联合甲钴胺治疗糖尿病周围神经病变的疗效及对血清SOD、MDA和AOPPs水平的影响

目的 探讨α-硫辛酸与甲钴胺联合治疗糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)患者的临床疗效及对氧化应激和炎性因子水平的影响。方法 回顾性分析74例DPN患者的临床资料,根据患者治疗方法的不同分为2组(对照组和实验组),对照组单用甲钴胺治疗,实验组接受α-硫辛酸和甲钴胺联合治疗; 治疗前后检测其氧化应激和炎症反应各指标水平的变化情况,同时对2组患者的临床疗效和不良反应进行评价。结果 所有受试者治疗后异常感觉、麻木等症状均明显改善(P<0.05),实验组治疗后各症状评分及总评分降低幅度明显大于对照组(P<0.05); 2组患者经过治疗后血清超氧化物歧化酶(SOD)、丙二醛(MDA)和(AOPPs)水平较治疗前均明显下降(P<0.05),且实验组降低更明显(P<0.05); 治疗后实验组血清肿瘤坏死因子(TNF-α)、C反应蛋白(CRP)和白细胞介素-6(IL-6)水平显著低于治疗前以及同期对照组(P<0.05); 2组患者恶心、恶吐等副作用发生率比较无显著差异(P=0.112)。结论 α-硫辛酸联合甲钴胺治疗DPN患者具有显著疗效,氧化应激及炎症反应明显改善,且不良反应发生率低     

Regulation of Th1/Th17 cytokines and IDO gene expression by inhibition of calpain in PBMCs from MS patients

Multiple sclerosis (MS) pathology is marked by the massive infiltration of myelin-specific T cells into the central nervous system (CNS). During active disease, pro-inflammatory Th1/Th17 cells predominate over immunoregulatory Th2/Treg cells. Here, we show that calpain inhibition downregulates Th1/Th17 inflammatory cytokines and mRNA in MS patient peripheral blood mononuclear cells (PBMCs) activated with anti-CD3/28 or MBP. Interestingly, calpain inhibition elevated IDO gene expression in MS PBMCs, which was markedly decreased in calpain expressing cells. Functional assay showed that incubation of MS patient PBMCs with calpain inhibitor or recombinant IDO attenuates T cell proliferation. These results suggest that calpain inhibition may attenuate MS pathology and augment the efficacy of standard immunomodulatory agents used to treat this disease.     

Sexual dimorphism in Th17/Treg axis in lymph nodes draining inflamed joints in rats with collagen-induced arthritis

Collagen type II-induced arthritis (CIA) in Dark Agouti rats, a model of rheumatoid arthritis (RA), reproduces sexual dimorphism in the incidence and severity of the human disease. Th17 cells are central in the induction/propagation of autoimmune inflammation in CIA and RA. To assess mechanisms underlying this dimorphism in CIA rats, in lymph nodes draining inflamed joints and adjacent tissues (dLNs) from CIA rats of both sexes Th17/CD25+Foxp3+CD4+ T-regulatory cell (Treg) ratio, Th17 cell redifferentiation in functionally distinct subsets and Treg transdifferentiation into IL-17-producing cells (exTregs) were examined. In female rats (developing more severe CIA than their male counterparts) the higher frequency of all Th17 cells (reflecting partly their greater proliferation), followed by the higher frequency of highly pathogenic IFN-γ/GM-CSF-co-producing cells, but lower frequency of less pathogenic/immunoregulatory IL-10-producing cells among them was found. Additionally, compared with male rats, in female rats the lower frequency of Tregs was observed. Moreover, Tregs from female rats exhibited diminished proliferative and suppressive capacity (judging by PD-1 expression) and enhanced conversion into IL-17-producing cells. Given that TGF-β concentration was comparable in collagen-type II-stimulated dLN cell cultures from female and male rats, the shift in Th17/Treg ratio followed by augmented Th17 cell redifferentiation into IFN-γ/GM-CSF-co-producing cells and Treg transdifferentiation into IL-17-producing cells in female rats was associated with increased concentration of IL-6 in female rat dLN cell cultures, and the higher frequency of IL-1β- and IL-23-producing cells among their dLN cells. The lower frequency of IL-10-producing B cells, presumably B regulatory cells (Bregs) could also contribute to the shift in Th17/Treg ratio in female rat compared with male rat dLNs. Consistently, the lower expression of IL-35 (the cytokine promoting Treg expansion directly and indirectly, by favoring Breg expansion and conversion into IL-10/IL-35-producing cells) in female rat dLN cells was detected. Thus, the study identified putative cellular and molecular substrates of the sexual dimorphism in the immunopathogenesis and clinical outcome of CIA and suggested mechanisms to be targeted in females to improve control of Th17 response, and consequently clinical outcome of CIA, and possibly RA.     

重症肌无力患者血清中miR150-5p与IL-10、IL-17水平的相关性研究

目的 检测miR150-5p在重症肌无力患者血清中的表达水平并分析与炎症因子水平的相关性。方法 收集符合本研究要求的重症肌无力患者62例和体检的健康志愿者30例,实时荧光定量聚合酶链反应(qRT-PCR)检测患者血清中miR150-5p的相对表达水平; 用酶联免疫吸附试验(ELISA)检测患者血清中炎症因子的水平; 采用pearson相关性分析法分析miR-150-5p与炎症因子水平的相关性。结果 重症肌无力患者血清中miR-150-5p的相对表达水平明显高于对照组(P<0.001); 重症肌无力患者血清中IL-2和IL-17的水平低于对照组(P<0.01),IL-10的水平高于对照组(P<0.01),而IL-19、IL-20和IL-35的水平与对照组比较无明显差异(P>0.05); 重症肌无力患者血清中miR-150-5p与IL-10的水平呈正相关(r=0.891,P<0.001),与IL-17的水平呈负相关(r=-0.836,P<0.001); 经过药物治疗后重症肌无力患者血清中miR-150-5p和IL-10的水平有所降低(P<0.001),而IL-2和IL-17的水平有所升高(P<0.001)。结论 重症肌无力患者中miR-150-5p的相对表达水平增加,且与IL-10的水平呈正相关,IL-17的水平呈负相关     

Th1 shift in CIDP versus Th2 shift in vasculitic neuropathy in CSF

To investigate the intra- and extracellular levels of various cytokines and chemokines in CSF in chronic inflammatory demyelinating polyneuropathy (CIDP) and vasculitic neuropathy (VN), 16 cytokines, IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IL-17, IFN-gamma, TNF-alpha, G-CSF, MCP-1 and MIP-1beta, were measured in CSF supernatant by a multiplexed fluorescent bead-based immunoassay and intracellular production of IFN-gamma and IL-4 in CSF CD4+ T cells were simultaneously measured by flow cytometry in 14 patients with CIDP, 8 patients with VN and 25 patients with other noninflammatory neurologic diseases (OND). In the CSF supernatant, a significant increase of IL-17, IL-8 and IL-6, and a significant decrease of IL-4, IL-5 and IL-7 levels were detected in pretreated CIDP as compared with OND. A significant increase of IL-6, IL-8 and IL-10 levels was found in pretreated VN. Both IL-17 and IL-8 levels correlated strongly with CSF protein levels in CIDP, although the correlation of IL-6 levels was weak. In CSF CD4+ T cells, IFN-gamma+ IL-4- cell percentages were markedly elevated in CIDP compared with OND, but not in VN, resulting in a significant increase of intracellular IFN-gamma/IL-4 ratio in CIDP, even in the absence of CSF pleocytosis. The nonresponders to intravenous immunoglobulins (IVIGs) showed a significantly lower IFN-gamma- IL-4+ CD4+ T cell percentage, and tended to have a higher intracellular IFN-gamma/IL-4 ratio than the responders in CSF. Marked upregulation of Th1 cytokine, IL-17, and downregulation of Th2 cytokines, together with infiltration of IFN-gamma-producing CD4+ T cells are useful markers for CIDP, while several Th2 cytokines are upregulated in VN in CSF.