Anti-inflammatory Effect of Polysaccharide from Embelia parviflora Wall.on Rheumatoid Arthritis in Rats

[Objectives]To investigate the anti-inflammatory effect of Embelia parviflora Wall.polysaccharide on rheumatoid arthritis(RA)in rats.[Methods]RA rat model was induced by type II collagen.After successful modeling,the rats were divided into model group,positive group,low,medium and high dose of E.parviflora Wall.polysaccharide groups,and normal control group.Body mass,toe volume and arthritis index were measured,and thymus index and spleen index were calculated.The levels of interleukin-1β,interleukin-6 and tumor necrosis factor-αin serum and synovial tissue of ankle joint were detected by ELISA.[Results]Compared with the normal control group,the pathological changes such as synovial hyperplasia and unclear layer were observed in the model group,the body mass was decreased(P<0.05),the toe volume,arthritis index,thymus and spleen index were increased(P<0.05),and the levels of IL-1β,IL-6 and TNF-αin serum and ankle synovial tissue were increased(P<0.05).Compared with the model group,the histopathological changes in synovium of ankle joint in the positive group and the medium and high dose groups of E.parviflora Wall.polysaccharide were significantly improved,and the body mass was increased(P<0.05).The toe volume,arthritis index,thymus index and spleen index were decreased(P<0.05).The levels of IL-Iβ,IL-6 and TNF-αin serum and synovial tissue of ankle joint were increased(P<0.05),while there was no significant difference between the low dose group of E.parviflora Wall.polysaccharide and the model group(P>0.05).[Conclusions]E.parviflora Wall.polysaccharide can reduce the body's inflammatory response and improve RA,which may be related to the inhibition of the activation of inflammatory cytokines IL-1β,IL-6 and TNF-α.

Design and evaluation of a novel transdermal patch containing diclofenac and teriflunomide for rheumatoid arthritis therapy

The aim of this study was to design a compound transdermal patch containing diclofenac(DA)and teriflunomide(TEF)for the treatment of rheumatoid arthritis(RA).The various organic amines salts of DA were prepared and their forming was confirmed using DSC and FTIR.The percutaneous permeation of organic amines salt of DA was investigated in vitro using a two-chamber diffusion cell with excised rabbit skin as transdermal barrier.The formulation of the patch was optimized in terms of the concentration of percutaneous permeation enhancer and the loading dose of drugs.The pharmacokinetic behavior of the optimal formulation was studies in rabbits and the anti-inflammatory and analgesic effects of the optimal patch were evaluated with the adjuvant arthritis model in rats and the pain model in mice,respectively.The result showed that skin penetration of diclofenactriethylamine(DA-TEtA)salt was better than other organic amine salts.Based on previous study of our laboratory,teriflunomide-triethylamine(TEF-TEtA)significantly enhanced the skin permeation of TEF.10%of azone(AZ)was the best enhancer for the two drugs.The optimal patch formulation was composed of 2%of TEF-TEtA,6%of DA-TEtA and 10%of AZ.The cumulative permeated amount of DA-TEtA in vitro was comparable with that of the commercial diclofenac-diethylamine(DA-DEtA)patch.The absolute bioavailability of TEFTEtA was 42%,which could achieve the therapeutic drug levels.In animal study,the optimized compound patch containing DA-TEtA and TEF-TEtA displayed significant antiinflammatory and analgesic effect,which indicated the potential of the compound patch.

The diagnostic significance of glucose-6-phosphate isomerase (G6PI) antigen and anti-G6PI antibody in rheumatoid arthritis patients

Objective: To investigate whether glucose-6-phosphate isomerase (G6PI) antigen and anti-G6PI antibodies could be applied for the clinical diagnostic markers of rheumatoid arthritis (RA) and its associations with RA activity states. Methods: The levels of G6PI antigens and anti-G6PI Abs in sera from 176 RA patients in different states, 35 non-RA patients and 100 healthy donors and in synovia fluids from 33 patients and 11 non-RA patients were measured by ELISA. Results: The sensitivity and specificity of G6PI antigens in the RA patients were 75.0% and 93.3%, respectively. The levels of serum G6PI antigens in 176 RA patients were significantly higher than non-RA patients and the health controls. Especially, there was a significant difference between the active phase and the inactive phase in G6PI antigens levels. The levels of G6PI antigens in synovia fluid were also significantly higher in RA groups than in non-RA patients. With the values of the anti-G6PI Abs in sera, there were no marked differences among RA, non-RA patients and health controls. Also, there was no significant difference between the active phase and the inactive phase in RA patients. However, there were no significant differences of G6PI and anti-G6PI between RA patients and health controls in synovial fluid. Conclusions: G6PI is highly correlated with the activity states of RA, and could be applied for a clinical biomarker with high sensitivity and specificity for the diagnosis of RA.

Study on Anti-Rheumatoid Arthritis Effect of Lignum Xanthocerais Sorbifoliae Based on Network Pharmacology

[Objectives]To explore the anti-rheumatoid arthritis effect of Lignum Xanthocerais Sorbifoliae by network pharmacology method.[Methods]The chemical constituents and action targets of Lignum Xanthocerais Sorbifoliae were collected by Traditional Chinese Medicine Systems Pharmacology(TCMSP).Target genes of rheumatoid arthritis(RA)were obtained from TTD database.ClusterProfiler was used for gene ontology(GO)enrichment,and Cytoscape 3.7.2 software was used for Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment and screening of active component-target core network.[Results]There were 24 intersecting targets between active components and diseases.The analysis of GO enrichment and KEGG pathway enrichment indicated that the therapeutic effect of Lignum Xanthocerais Sorbifoliae on rheumatoid arthritis might be related to AGE-RAGE signaling pathway,IL-17 signaling pathway,rheumatoid arthritis pathway,C-type signaling pathway and Toll-like receptor signaling pathway.13 drug active components[flavanes,dihydroflavonoids,stigmasterol,oleanolic acid,kaempferol,ellagic acid,Terchebulin B,Copaene,(Z)-β-Caryophyllene,Aromadendrene,β-Elemene,emodin]and 12 core targets(TNF,IL6,ALOX5,MAPK14,PPARA,AKR1B1,CCR5,PLA2G1B,ADAM17,MMP1,PTGS1,SYK)constituted a core network.[Conclusions]This study could provide a theoretical basis for the treatment of RA with Lignum Xanthocerais Sorbifoliae preparation.

Increased IgG Rheumatoid Factor-Positivity in the Asian Rheumatoid Arthritis Patients Irrespective of Ethnicity

Aim of Work: Initial observations implied IgG rheumatoid factor (RF) to be common among Malaysian rheumatoid arthritis (RA) patients. We tested this hypothesis and used a multiethnic RA cohort (Malays, Chinese and Indians) to investigate whether the IgG RF predominance might be genetically or environmentally determined. Patients and Methods: 556 serum samples comprising 171 patients classified as RA according to the 1987 ACR criteria, 60 patients with other rheumatic diseases and 325 non-rheumatic controls were tested for IgG RF, IgM RF and anti-CCP by ELISA. The findings were then tested in a larger RA case-control cohort (n = 1844). Results: IgG RF predominated over IgM RF in all the investigated ethnic groups. The sensitivity, specificity, and diagnostic accuracy of IgG RF (55.6%, 91.2% and 80.2%, respectively) were superior compared to IgM RF, but comparable to anti-CCP. IgG RF was however, also increased in the Malaysian controls, but the IgG RF superiority over IgM RF was still apparent after cutoff adjustment according to the 1987 ACR criteria. Autoantibody levels did not differ between the three ethnic groups. The Receiver Operating Characteristics (ROC) curves showed larger areas under the curves for IgG RF (0.826) and for anti-CCP (0.867) than for IgM RF (0.737). Review of the literature showed consistently higher sensitivity for IgG RF in studies of Asian RA patients as compared to Caucasian and African-American studies. Conclusion: Increased frequency of IgG RF-positive in RA populations with different genetic background living in Malaysia argues for an environmental factor selectively amplifying the IgG RF response.

Diagnostic Value of Antibodies against a Modified Citrullinated Vimentin in Rheumatoid Arthritis

Aim of Work: To investigate the value of the detection of antibodies against modified citrullinated vimentin antibodies (anti-MCV) in comparison with anti-CCP2-for the diagnosis of rheumatoid arthritis (RA). Patients and Methods: The study Included Forty patients with Rheumatoid arthritis (RA). They under went assessment by the disease activity score (DAS-28), visual analogue scale (VAS) and health assessment questionnaire (HAQ). Thirty healthy subjects matched for age and sex served as a control group. Blood samples were obtained from patients and controls for erythrocyte sedimentation rate (ESR), C reactive protein (CRP), rheumatoid factor (RF). Anti-CCP2 and anti-MCV were determined using ELISA technique. Results: Estimated serum levels of anti-CCP2 and anti-MCV were significantly higher in patients compared to controls (p 0.001). There were no significant correlations between anti-MCV levels and age, dis- ease duration, duration of morning stiffness, number of swollen and tender joints, HAQ or ESR in patients with RA, while serum levels correlates significantly with DAS28, VAS and CRP (p 0.05). Anti-CCP2 correlates significantly with DAS28, VAS and CRP and ANA (p 0.05). Serum anti-MCV and anti-CCP2 were significantly correlated with each other (r = 0.483;p The receiver operating characteristic (ROC) curve was drawn and it showed that anti-MCV had diagnostic specificity, sensitivity of 93.3%, 75.5%, respectively, while anti-CCP2 specificity, sensitivity of 98.1%, 85%, respectively. Conclusion: Serum anti-MCV as well as the anti-CCP-2 assay perform comparably well in the diagnosis of RA. In the high-specificity range, however, the anti-CCP2 assay appears to be superior to the anti-MCV test.

Meta-Analysis on Efficacy and Safety of Modified Guizhi Shaoyao Zhimu Tang with the Western Medicines in Treating Rheumatoid Arthritis

Objective: To assess the efficacy and safety of modified Guizhi Shaoyao Zhimu Tang (GSZT) with the Western medicines in treating rheumatoid arthritis. Methods: All the relevant studies were collected by searching the Pubmed, Web of science, Embase, Cochrane Library, SpringerLink, China National Knowledge Infrastructure (CNKI), VIP Journals database, and Wanfang database from their inception to October 2017. After the assessment, the eligible ones were subject to Jadad score. The meta-analysis was conducted by RevMan 5.3 software. Results: Totally 10 randomized controlled trials (RCTs) involving 962 patients were enrolled. The clinical efficacy of analysis showed that modified GSZT with the Western medicines can significantly improve the clinical total effective rate [OR = 4.64, 95%CI (3.03, 7.12), P &minus;14.58, 95%CI (&minus;16.49, &minus;12.68), P &minus;12.62 95%CI (&minus;18.69, &minus;6.54), P &minus;49.88, 95%CI (&minus;77.6, &minus;22.09), P &minus;5.34, 95%CI (&minus;5.86, &minus;4.82), P < 0.001]. The adverse events had been seen in both of the two groups, however, the clinical symptoms of the group on modified GSZT with the Western medicines were obviously reduced than the other one [OR = 0.25, 95%CI (0.10, 0.61), Z = 3.06, P < 0.01]. This system review shows that the modified GSZT with the Western medicines improved the clinical efficacy, symptoms and the blood biochemical indicators were improved significantly than that of Western medicines alone on rheumatoid arthritis. At the same time, it can reduce adverse effects and enhance safety.

艾灸对实验性RA大鼠海马下丘脑垂体GR表达影响的研究

目的:在观察艾灸对实验性RA大鼠抗炎效应的基础上,研究艾灸对海马、下丘脑及垂体GR表达的影响,探索艾灸治疗RA的GR机制。方法:将雄性SD大鼠随机分为空白对照组、模型组、艾灸组,用福氏完全佐剂塑造大鼠实验性RA病理模型,艾灸"肾俞"、"足三里"治疗,测定大鼠右后足容积,采用免疫组化法检测海马、下丘脑、垂体GR表达。结果:造模后模型组大鼠足肿胀度明显高于空白对照组(P<0.01),艾灸组大鼠足肿胀度明显低于模型组(P<0.01);海马GR表达,模型组与空白组比较有增高趋势(P>0.05),艾灸组明显低于模型组(P<0.01);下丘脑GR表达,模型组明显高于空白对照组(P<0.01),艾灸组较模型组明显降低(P<0.01);垂体GR表达,模型组较空白对照组显著升高(P<0.01),而艾灸组与模型组比较无显著性差异(P>0.05)。结论:艾灸对实验性RA具有抗炎效应,艾灸对RA大鼠海马、下丘脑GR表达的调控可能是其实现抗炎效应的重要机制。

抗CCP、抗AKA和抗RA33抗体检测在类风湿性关节炎诊断中的应用

目的探讨环瓜氨酸多肽(CCP)抗体、抗角蛋白抗体(AKA)、类风湿因子(RF)和抗RA33抗体对类风湿性关节炎(RA)的临床应用价值。方法88例RA患者和50例其他自身免疫病患者分别采用ELISA法检测抗CCP和抗RA33抗体,免疫荧光法检测抗AKA抗体,速率散射比浊法检测类风湿因子(RF),观察各指标对RA的诊断价值。结果患者抗CCP抗体、抗AKA抗体、抗RA33抗体和RF的敏感度分别为68.2,28.4,39.8和71.6,特异度为92.0,100,94.0和70.0;抗CCP抗体的检测阳性率高于抗AKA抗体和抗RA33抗体(P<0.01),与RF无统计学显著性差异(P=0.622)。三种抗体对RF阴性组RA诊断的阳性率为63.0,37,44.4,与RF阳性组RA阳性率70.5,24.6,37.7相比较,两组差异无统计学意义(P>0.05)。结论抗CCP,抗AKA,抗RA33抗体均是RA诊断的特异性指标,对诊断RF阴性RA具有较高敏感度和特异度,将上述抗体与RF进行联合检测,有助于RA的早期诊断。

艾灸调整实验性RA大鼠HPAA功能的GR机制研究

目的:探索艾灸调整实验性类风湿性关节炎(RA)大鼠下丘脑-垂体-肾上腺轴(HPAA)功能的糖皮质激素受体(GR)机制。方法:将SD大鼠随机分为空白对照组、模型组、艾灸组、艾灸加注射RU28362组,用福氏完全佐剂塑造实验性RA病理模型,以GR激动剂RU28362腹腔注射作为干预研究手段,艾灸"肾俞"、"足三里"穴治疗,检测大鼠足容积,用ELISA法测定大鼠血浆CRH、ACTH、GC含量。结果:模型组大鼠足肿胀度明显高于空白对照组(P<0.01),艾灸组大鼠足肿胀度明显低于模型组(P<0.01),艾灸加注射RU28362组足肿胀度与模型组比较无显著性差异(P>0.05);与空白组对照组比较,模型组血浆CRH、GC含量显著增高(P<0.01),ACTH含量显著降低(P<0.05);艾灸组血浆CRH、GC含量明显低于模型组(P<0.01),ACTH含量明显高于模型组(P<0.05),但艾灸加注射RU28362组血浆CRH、ACTH、GC含量与模型组比较无显著性差异(P>0.05)。结论:艾灸对实验性RA具有明显的抗炎效应;艾灸对实验性RA大鼠的HPAA功能紊乱有良性调整作用;海马GR在艾灸调整实验性RA大鼠HPAA功能中具有重要作用。

不同刺灸法调节RA细胞因子的实验研究进展

细胞因子水平异常是RA重要发病机理,针灸治疗RA疗效显著,研究表明,针灸良性调节RA异常的细胞因子是发挥治疗效应的关键所在。笔者检索了近10年相关文献并整理、综述、分析不同刺灸法对RA发生特异改变的不同细胞因子的影响。在此基础上通过比较阐释针灸治疗RA的有效方法,并评述该领域实验研究存在的问题,为今后深入研究提供参考和依据。

辅助T细胞多巴胺受体(DR)表达对类风湿关节炎(RA)患者疾病活动度的影响

目的 检测类风湿关节炎（rheumatoid arthritis,RA）患者外周血辅助T淋巴细胞（T helper cells,Th cells）不同亚群细胞膜表面多巴胺受体（dopamine receptor,DR）水平,分析其与疾病活动度、实验室指标及功能状态之间的关系。方法 选取25位初诊或3个月内未接受DMARDs治疗的RA患者（DAS28〉3.2）,12位健康对照及13位同期年龄匹配的骨关节炎（osteoarthritis,OA）患者。采用流式细胞术检测患者外周血Th细胞亚群表面DR1~DR5的表达情况,分析Th细胞DR表达水平与人口学资料、实验室检查、功能状态及疾病活动度之间的相关性。结果 RA组Th2细胞百分比为1.28%（0.74%~2.68%）,较健康对照组（中位数0.18%,四分位数间距0.11%~0.31%,P〈0.001）及OA组（中位数0.48%,四分位数间距0.03%~1.47%,P=0.029）明显升高。RA组Th2细胞DR2表达水平为25.40%（四分位数间距3.79%~37.10%）,明显高于健康对照组（中位数3.74%,四分位数间距0.00%~12.98%,P=0.006）及OA组（中位数3.45%,四分位数间距0.00%~20.55%,P=0.040）。RA组Th2细胞DR4表达量也高于健康对照组和OA组（健康对照组vs.RA组,P=0.022;OA组vs.组,P=0.010）。Th2细胞与关节肿胀数（swollen joint count,SJC）和简化疾病活动指数（SDAI）呈正相关（分别为r=0.421,P=0.036;r=0.396,P=0.050）。Th1细胞DR5百分比分别为2.22%（四分位数间距0.57%~12.24%）与SJC及临床疾病活动指数（CDAI）呈正相关（分别为r=0.492,P=0.012;r=0.445,P=0.026）。Th2细胞DR1和DR3百分比分别为11.15%（四分位数间距2.93%~36.20%）、9.80%（1.62%~23.05%）,均与健康评估问卷（Health Assessment Questionnaire,HAQ）评分呈负相关（分别为r=-0.469,P=0.018;r=-0.464,P=0.019）。结论 RA患者Th2细胞多巴胺受体DR2和DR4明显升高,Th细胞DR水平与RA患者SJC、CDAI及HAQ存在相关性。

类风湿性关节炎与IL-1/IL-1Ra平衡关系的研究进展

类风湿性关节炎(RA)为常见的自身免疫疾病,其发生、发展与众多炎性因子相关,白细胞介素1(IL-1)是一种炎症反应的重要介质,在人体的自然反应和病理条件导致慢性炎症的发展中都有重要作用,然而在抗炎细胞因子的影响下,体外IL-1炎症作用可以被一个特别强大的抑制剂——IL-1受体拮抗剂(IL-1Ra)抑制或者消除。目前研究显示,IL-1通过与Ⅰ型IL-1受体结合,可激活系列信号转导通路进而诱发关节软骨和骨侵蚀,同时可诱导其他炎性因子表达,协同加剧关节破坏,而IL-1Ra可牢固结合Ⅰ型IL-1受体,抑制IL-1的生物学效应,因此,RA患者体内IL-1/IL-1Ra的动态平衡对于该病的转归至关重要。深入对RA发病中IL-1/IL-1Ra平衡的认识对该病的预防、治疗及转归具有重要作用。

清络通痹颗粒对RA患者外周血T淋巴细胞表达RANKL的影响

目的:观察清络通痹颗粒(QLT)对类风湿关节炎(RA)患者外周血T淋巴细胞表达破骨细胞分化因子(RANKL)的影响。方法:采用流式细胞术检测RA患者及健康志愿者外周血T淋巴细胞表达RANKL情况;同时检测了添加清络通痹颗粒含药血清后RA患者外周血T淋巴细胞表达RANKL的水平。结果:活动期RA患者外周血T淋巴细胞RANKL表达率显著高于健康者(P<0.01);添加清络通痹颗粒含药血清后RA患者外周血T淋巴细胞RANKL表达的水平明显下降(P<0.05或<0.01)。结论:清络通痹颗粒有可能通过下调T淋巴细胞RANKL的表达,从而抑制破骨细胞(OC)的分化。

检测早期类风湿关节炎(RA)患者血清中IL-33水平的临床意义

目的观察探对检测早期类风湿关节炎(RA)患者血清中的IL-33水平的结果,总结其检测的临床意义。方法选取我院2009年7月至2011年7月收治的早期类风湿关节炎(RA)患者48例作为观察组,以同期在我院进行健康体检者50例作为对照组,观察比较两组受检者血清中IL-33水平,同时也检测观察组患者滑液及血清中的IL-33水平。结果观察组患者的血清中IL-33水平明显高于健康者(P<0.05),具有统计学意义;观察组患者的滑液和血清中均有IL-33表达,但浓度比较无显著差异(P>0.05),无统计学意义。结论对早期类风湿关节炎(RA)患者进行血清IL-33水平的检测,能够较好地反应出滑膜内的病变严重程度,对早期诊断并评估类风湿关节炎(RA)患者的疾病发展有重要的临床指导意义。

中药熏蒸方对佐剂性关节炎大鼠IL-1Ra/IL-1β的影响

目的:研究中药熏蒸方对佐剂性关节炎(adjuvant arthritis,AA)大鼠血清、关节滑膜组织中的白细胞介素1β(IL-1β)、IL-1受体拮抗剂(IL-1Ra)的影响,并探讨其改善类风湿性关节炎(RA)的作用机制。方法:采用弗氏完全佐剂诱导AA大鼠模型,随机分为模型组、MTX组、中药熏蒸组、MTX+中药熏蒸组,分别给予中药熏蒸,MTX(1.9 mL·kg^(-1)·w^(-1))腹腔注射,中药熏蒸+MTX(1.9 mL·kg^(-1)·w^(-1))腹腔注射,另设正常大鼠为正常组,连续给药20天后处死大鼠收集血清、关节滑膜组织。观察大鼠关节肿胀度和关节炎指数的变化,酶联免疫吸附剂(ELISA)测定检测血清IL-1β、IL-1Ra的水平,反转录聚合酶链式反应(RT-PCR)法检测滑膜组织中IL-1β、IL-1Ra mRNA表达量的变化,并观察中药熏蒸方对这些指标的影响。结果:与正常组比较,模型组关节肿胀度和关节炎指数明显升高,血清、滑膜组织IL-1β表达水平明显升高、IL-1Ra表达水平下降;中药熏蒸方能抑制模型组血清、滑膜组织IL-1β的表达,上调血清、滑膜组织IL-1Ra表达量,而中药熏蒸方和MTX两药协同的效果更加明显。结论:中药熏蒸方改善AA大鼠关节炎机制可能与恢复IL-1Ra/IL-1β的平衡相关。

淋巴细胞共刺激信号和凋亡信号异常与RA免疫效应亢进的研究

目的探究T淋巴细胞表面多种细胞信号分子所介导的细胞活化或凋亡信号在RA患者免疫功能紊乱中的作用。方法采用流式细胞术检测RA患者外周血T细胞亚群及其表面共刺激分子CD154(CD40L)、CD30和凋亡受体CD95(Fas)的表达。结果RA患者外周血T细胞亚群偏移,CD4+T细胞增加,CD8+T细胞减少;共刺激分子CD154在CD4+和CD8+T细胞上的表达均上调,但CD30分子的表达均降低,并以CD4+T细胞降低更为明显。同时,凋亡受体CD95分子在T细胞亚群上的表达均明显增加。结论RA患者T淋巴细胞表面多种信号分子表达异常,共同导致了RA患者免疫功能紊乱。

类风湿关节炎(RA)患者Th1、Th2型细胞在患者病情演变中的特点及临床意义

目的观察探讨类风湿关节炎(RA)患者Th1、Th2型细胞在患者病情演变中的特点,总结其临床意义。方法选取51例类风湿关节炎患者为观察组,分别在其治疗前和治疗后3个月进行Th1、Th2型细胞检测,再选取健康体检者60例为对照组,也进行同样的细胞检测,观察比较两组治疗前后的差异性。结果观察组治疗前与对照组比较,Th1型细胞显著高于对照组、Th2型细胞低于对照组,差异有统计学意义(P<0.05),观察组患者经过治疗后,其血清中Th1型细胞明显降低,而Th2型细胞明显升高,与治疗前比较有统计学意义(P<0.05)。结论对类风湿性关节炎患者进行Th1、Th2型细胞检测,能有助于判断患者的病情进展,对评价临床疗效及预测预后有重要的临床指导意义。

含噻唑烷-4-酮的免疫调节剂对RA患者iNKT细胞免疫调节功能的影响

目的:研究新型合成的含噻唑烷-4-酮的免疫调节剂(CH1b)对活动期RA患者iNKT细胞免疫调节功能的影响。方法:从RA患者外周血中分离得到单个核淋巴细胞(PBMC),α-Galcer和IL-2体外刺激扩增后,经磁珠分选(MACS)得到纯化的iNKT细胞,分成对照组(IL-2)、α-Galcer组(IL-2+α-Galcer)、CH1b组(IL-2+CH1b)。采用MTT法测定各组RA患者iNKT细胞增殖率;MILLIPLEX MAP Human Cytokine/Chemokine kit检测各组RA患者iNKT细胞培养上清中IFN-γ和IL-4的水平;采用RT-PCR方法检测各组RA患者iNKT细胞中IFN-γmRNA和IL-4 mRNA的表达水平。结果:与对照组和α-Galcer组相比,CH1b组iNKT细胞增殖率明显增加(P<0.05);与对照组相比,α-Galcer组IFN-γ和IL-4分泌水平明显增加(P<0.05),CH1b组IFN-γ和IL-4的分泌量也明显增加,IFN-γ/IL-4明显降低(P<0.05);与对照组相比,α-Galcer组IFN-γmRNA与IL-4 mRNA表达均明显增高,CH1b组IFN-γmRNA表达差异不明显,IL-4 mRNA的表达明显增高。结论:新型合成的免疫调节剂(CH1b)可促进活动期RA患者活化的iNKT细胞增殖,并促进IL-4的分泌,上调IL-4 mRNA的表达,诱导iNKT细胞向Th2方向分化,对恢复RA病人体内免疫平衡具有重要意义。

Ras-MAPKs信号通路在类风湿关节炎miRNA-146a调控DNMTs中的作用及温化蠲痹方干预作用研究

目的研究ras-MAPKs信号通路在类风湿关节炎(rheumatoid arthritis,RA)患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)微小RNA-146a(miRNA-146a)对DNA甲基化转移酶(DNA methyltransferase,DNMTs)调控中的作用,以及温化蠲痹方干预作用。方法提取RA患者、健康受试者PBMCs进行体外培养研究。将18例健康受试者随机分为6组,每组均为3例,分别为正常组,正常转染miRNA-146a组(转染组),正常转染miRNA-146a加中药组(中药1组),正常转染miRNA-146a加MAPKs特异性阻断剂PD98059组(阻断1组)、SB203580组(阻断2组)、SP600125组(阻断3组)。将符合中医辨证为寒热错杂、痰瘀痹阻型的活动期RA患者15例,随机分为5组,每组3例,分别为RA组,RA加中药组(中药2组),RA加MAPKs特异性阻断剂PD98059组(阻断4组)、SB203580组(阻断5组)、SP600125组(阻断6组)。对受试者PBMCs进行体外培养、转染、加通路阻断剂以及含药血清,运用实时定量PCR检测PBMC中miRNA-146a、DNMTs、RASGRP1的表达。结果阻断1~6组miRNA-146a表达降低,DNMTs表达升高,RASGRP1表达降低(均P<0.05)。中药1、2组RSAGRP1表达降低(均P<0.05)。结论ras-MAPKs信号通路可能参与了RA患者miRNA-146a对DNMTs的调控。中药温化蠲痹方可能通过下调RA患者RASGRP1表达作用于ras-MAPKs信号通路,影响miRNA-146a对DNMTs进行调控,达到治疗RA作用。