槲皮素对大鼠心肌梗死后左心室重塑的改善机制研究

目的 探讨槲皮素(Que)对大鼠心肌梗死(MI)后心室重塑的保护作用及对炎症细胞因子肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的影响。方法 将30只SD雄性大鼠随机分为假手术组(Sham组)、MI组和Que组,每组各10只。MI组采用结扎左前降支构建模型,Sham组只穿针不结扎,Que组在MI后予以槲皮素100 mg·kg^-1·d^-1喂养。喂养28 d后,予以心脏彩色多普勒超声评估心脏结构和功能,马松(Masson)染色测定心肌纤维化程度,ELISA法观察大鼠血液循环中TNF-α和IL-6水平,免疫组织化学染色(IHC)检测心脏组织中TNF-α和IL-6表达水平。结果 MI组较Sham组左心室收缩末期容量(LVESv)及左心室舒张末期容量(LVEDv)增大,左心室射血分数(LVEF)及左心室短轴收缩率(LVFS)下降,左心室心肌纤维化程度升高,循环中及心脏组织中TNF-α和IL-6表达水平升高,差异有统计学意义(P<0.05);Que组较MI组LVESv缩小,LVEF及LVFS上升,左心室心肌纤维化程度下降,循环中及心脏...     

生脉胶囊对慢性心力衰竭大鼠多种细胞因子的影响

目的探讨生脉胶囊对慢性心力衰竭大鼠细胞因子IL-6、TNF-α、IL-10、TGF-β1的影响。方法将大鼠随机分为假手术组、心衰模型组、生脉胶囊组、卡托普利组、生脉胶囊加卡托普利组,采用腹主动脉缩窄法造成大鼠慢性心力衰竭模型,治疗7周时采用ELISA法测定大鼠心肌IL-6、TNF-α、IL-10和血清TGF-β1的含量。结果①心衰模型组心肌IL-6、TNF-α、TGF-β1的含量均高于假手术组(P<0.05或P<0.01),生脉胶囊组、卡托普利组、生脉胶囊加卡托普利组含量均分别低于心衰模型组(P<0.01或P<0.05),但生脉胶囊加卡托普利组TGF-β1含量低于生脉胶囊组(P<0.05)。②心衰模型组心肌IL-10的含量明显低于假手术组(P<0.01),生脉胶囊组高于心衰模型组(P<0.05)。结论生脉胶囊可降低心肌炎性细胞因子IL-6、TNF-α的含量,增加抗炎细胞因子IL-10的含量,降低促纤维性细胞因子TGF-β1的含量,与抑制胶原的合成代谢、改善慢性心衰心室重构有关。     

银杏酮酯预处理对心肌缺血再灌注大鼠心肌组织炎症相关细胞因子含量的影响

目的：探讨银杏酮酯（Ginkgo biloba extract 50,GBE50）预处理对大鼠心肌缺血再灌注损伤的保护作用及对炎症细胞因子和抗炎因子的影响。方法：58只SD大鼠分为假手术组,模型组,丹参组和低、中、高剂量GBE50组（n=8）。灌胃药物1周后,结扎冠状动脉左前降支30 min,再灌60 min建立大鼠心肌缺血再灌注模型。取心肌组织,HE染色观察心肌组织形态学变化,比色法检测心肌组织中髓过氧化物酶（myeloperoxidase,MPO）活性,放射免疫法检测心肌组织中肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）、白细胞介素6（interleukin-6,IL-6）和IL-8含量,酶联免疫吸附测定法检测心肌组织中IL-4和IL-10含量。结果：与模型组相比,中剂量GBE50组心肌炎症反应减轻,心肌组织MPO活性明显降低（P〈0.01）;GBE50可明显减少心肌IL-6含量（P〈0.05,P〈0.01）,升高IL-4含量（P〈0.05,P〈0.01）。与模型组比较,GBE50组心肌组织中TNF-α含量降低,IL-10含量升高,但差异无统计学意义。结论：GBE50可降低缺血再灌注后心肌IL-6含量,升高IL-4含量,提示其可能通过抑制炎性细胞因子、促进抗炎因子的表达,对心肌缺血再灌注后的炎症反应发挥调控作用。     

油酸-内毒素序贯致伤大鼠血浆和肺泡灌洗液TNF-α、IL-1β和IL-6水平的变化

目的探讨促炎症细胞因子:肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)水平的变化,在全身炎症反应和急性肺损伤中的作用.方法采用油酸加内毒素脂多糖(LPS)序贯致伤大鼠,在油酸致伤后1、4、12和24h实施小剂量LPS第2次致伤,收集血浆和肺泡灌洗液,采用酶联免疫法(ELISA)检测TNF-α、IL-1β和IL-6蛋白含量.以单油酸致伤和单小剂量LPS致伤作为对照组进行比较分析.结果 TNF-α和IL-1β在油酸致伤4 h LPS第2次致伤后水平最高,IL-6的最高水平在油酸12 h加LPS致伤组;2次序贯致伤后IL-1β水平变化最大,增高最显著,血浆浓度达到(1 526±39.69)pg/ml,肺泡灌洗液内浓度为(840.4±95.29)pg/ml.结合肺组织病理学研究,肺组织损伤最严重大鼠,血浆和肺泡灌洗液TNC-α、IL-1β和IL-6水平最高.结论 TNF-α和IL-1β是早期释放的促炎症细胞因子,其中IL-1β的血浆和肺泡灌洗液中水平最高,它可能在全身炎症反应发生、发展和急性肺损伤中起非常重要的作用.肺组织损伤程度与这些促炎症细胞因子水平有密切关系.     

中长期使用特异性COX-2抑制剂对大鼠左心室重塑的影响

目的 通过观察大鼠慢性压力负荷性心肌肥厚过程中左心室重塑2个时间点使用特异性COX-2抑制刑Celecoxib前后左心室质量指数(LVMI)、心脏重/体重比值和炎症标记物的变化,以了解中长期使用特异性COX-2抑制剂对左心室重塑的作用.方法 将40只雄性SD大鼠随机分成5组:假手术组、手术20周组、Cele-coxib 20周组、手术24周组、Celecoxib 24周组.手术组及Celecoxib组均按照腹主动脉缩窄法制作压力负荷性心肌肥厚大鼠模型,在造模16周后,用Celecoxib 10 mg/kg,均匀混入饲料中喂服Celecoxib组大鼠,分别连续喂养4周和8周.在造模20周及24周时,观察大鼠左心室质量指数和炎症标记物TNF-α、TGFβ-β、PGE2、CRP、UA变化.结果 手术组20和24周LVMI、TNF-α、PGE2、CRP、UA较假手术组均有升高(P<0.01,P<0.05);Celecoxib24周组LVMI及心脏重/体重比值、TNF-α、TGF-β、PGE2、CRP、UA均较手术组有下降(P<0.01,P<0.05).结论 炎症参与了心肌重塑的过程,特异性COX-2抑制剂Celeeoxib可能通过抗细胞因子起到延缓左心室重塑发展.     

辛伐他汀对压力超负荷大鼠APN、IL-6及心肌肥厚影响

目的 观察辛伐他汀对压力超负荷大鼠血清脂联素(APN)、白细胞介素-6(IL-6)水平以及心肌肥厚的影响,探讨该类药物影响心肌肥厚可能的作用机制.方法 实验动物随机分对照组、心肌肥厚组、治疗组,每组10只.应用经典的腹主动脉结扎方法,制备压力超负荷大鼠模型.治疗组大鼠给予辛伐他汀5 mg/kg 每日清晨灌胃,连续6周.心肌肥厚组和对照组同时以同等剂量的9 g/L的氯化钠溶液每日清晨灌胃,直至术后6周.采用鼠尾测压法测定大鼠血压的变化;酶联免疫吸附试验法检测大鼠血清APN、IL-6浓度.实时定量PCR检测心肌组织中IL-6、脂联素受体1(AdipoR1)的mRNA表达.称量法计算各组大鼠心脏质量及心脏质量/体质量比值,并观察心肌的病理学改变.结果 心肌肥厚组大鼠血清APN水平、IL-6水平、心脏质量、心脏质量/体质量比值、左心室壁平均厚度、心肌细胞平均直径,以及心肌IL-6、AdipoR1 mRNA表达与对照组相比差异有显著性(q=3.41～31.83,P<0.01).治疗组大鼠与心肌肥厚组相比血清APN水平、IL-6水平、心脏质量、心脏质量/体质量比值、左心室壁平均厚度、心肌细胞平均直径以及心肌IL-6、AdipoR1 mRNA表达差异有显著性(F=3.57～329.93,q=3.11～31.08,P<0.01).结论 辛伐他汀可能通过增加血清APN的浓度以及心肌细胞因子的表达来抑制压力负荷增加引起的大鼠心肌肥厚.     

AT2参与调控肥大心肌细胞炎症因子的合成和分泌

目的 观察血管紧张素Ⅱ二型受体(AT2)对压力超负荷性肥大心肌细胞炎症因子合成和分泌的影响.方法 采用腹主动脉缩窄法构建成年SD大鼠压力超负荷性心肌肥大模型,选用缩窄术后8周的肥大心肌细胞,经Ang Ⅱ及losartan或PDl23319处理36 h后.放免法检测培养上清液中IL-1β、TNF-α及IL-6的表达水平.结果 随着浓度增大,AngⅡ可以诱导肥大心肌细胞TNF-α和IL-1β分泌.PDl23319,而不是losartan,预处理可使TNF-α和IL-1β表达下调.实验中未能检测出培养液中IL-6含量.结论 AngⅡ诱导压力超负荷性肥大心肌细胞炎症因子TNF-a和IL-1β的合成与释放主要受AT2的调控.     

内毒素耐受对细胞因子的影响

目的进一步明确内毒素耐受时炎性相关细胞因子的变化。方法昆明鼠分组处置。建立内毒素(LPS)耐受鼠模型。ELISA方法测定血清肿瘤坏死因子α(TNF-α),白细胞介素10(IL-10)和白细胞介素17(IL-17)浓度。结果 LPS对照组血清TNF-α、IL-10和IL-17浓度分别为(2 261.49±1 090.28)pg/m L,(7 780.42±2 320.04)pg/m L,(27.28±13.76)pg/m L,较健康对照组显著增高。而内毒素耐受组相应细胞因子血清浓度分别为(1 174.19±450.91)pg/m L,(1 293.26±520.85)pg/m L,(22.26±11.44)pg/m L。地塞米松干预进一步降低了TNF-α和IL-17的水平,但对IL-10水平无显著影响。结论内毒素耐受状态与内毒素炎症状态比较,TNF-α和IL-10水平显著降低,IL-17水平无显著变化。应用地塞米松可进一步降低内毒素耐受状态下的TNF-α和IL-17水平。     

压力超负荷心肌肥厚过程中血清TNF-α、IL-1 β、IL-6的变化

目的:探讨从压力负荷增加到心肌肥厚形成的整个过程中血清肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)的变化及意义.方法:采用放射性免疫法检测腹主动脉部分结扎大鼠不同时间点血清TNF-α、IL-1β、IL-6的浓度,电子天平称量左心室重,计算心重指数(左心重mg/体重g).结果:与对照组相比,心肌肥厚大鼠血清TNF-α、IL-1β、IL-6升高,且与心肌肥厚程度正相关.结论:细胞因子可能参与了心肌肥厚的发生和发展.     

长效奥曲肽对肝纤维化大鼠IL-6、IL-8的影响

目的 观察长效奥曲肽对四氯化碳(CCl4)诱发大鼠肝纤维化白细胞介素-6(interleukin-6,IL-6)、白细胞介素-8(interleukin-8,IL-8)的影响.方法 SD大鼠40只,随机分为正常对照组(n=8)、肝纤维化组(n=16)和长效奥曲肽组(n=16).以40%CCl4皮下注射(3mL/kg)建立大鼠肝纤维化模型,长效奥曲肽组同时给予长效奥曲肽(0.8mg/kg),每4周肌肉注射1次.8周后观察肝脏组织形态学改变、用酶联免疫吸附法(ELISA)检测血清IL-6和IL-8.结果 正常对照组大鼠肝脏大体形态和组织学无异常改变;肝纤维化门脉高压组及长效奥曲肽组大鼠肝脏组织形态学表现为肝纤维化改变,但后者的病理损害指标显著轻于前者(P<0.05);正常对照组血清IL-6、IL-8含量分别为(175.28±31.15)pg/mL、(81.51±16.48)μg/mL,肝纤维化组血清IL-6、IL-8含量分别为(313.27±52.79)pg/mL、(213.15±31.16)μg/mL,而长效奥曲肽组分别为(265.13±46.57)pg/mL、(185.16±32.56)μg/mL,肝纤维化组和长效奥曲肽组血清IL-6、IL-8均显著高于正常对照组,但后者两项指标显著低于前者(P<0.05).结论 长效奥曲肽能减轻大鼠肝纤维化程度及降低血清IL-6和IL-8水平.     

白介素-1受体拮抗剂对缺血再灌注心肌的保护作用及其机制探讨

目的　探讨急性心肌梗死 (AMI)再灌注过程中白介素 - 1β(IL 1β)的变化及重组人白介素 - 1受体拮抗剂 (rhIL 1ra)对缺血再灌注心肌的保护作用及其机制。方法　 (1)用ELISA法测定了 8名健康人和 2 2例接受急诊经皮冠脉介入治疗 (PCI)并且梗死相关血管 (IRA)血流达心肌梗死溶栓 (TIMI) 3级的AMI患者血浆IL 1β水平的动态变化。 (2 )在兔心肌缺血 5 0min再灌注 4h的模型上 ,观察不同剂量rhIL 1ra(A组 :rhIL 1ra 10mg/kg ;B组 :rhIL 1ra 2 0mg/kg ;C组 :rhIL 1ra 4 0mg/kg)对心肌梗死面积、心肌髓过氧化物酶 (MPO)活性、心肌细胞凋亡程度 (TUNEL、AnnexinⅤ和DNA 梯形图法 )及凋亡相关基因Bcl 2 /Bax的表达。结果　 (1) 2 2例AMI患者再灌注前血浆IL 1β(2 8pg/ml±9pg/ml)已明显高于健康对照组 (2 0 pg/ml± 11pg/ml,P <0 0 5 ) ,IRA再通后 12h血浆IL 1β水平达高峰 (86 pg/ml± 14pg/ml) ,至术后 2 4hIL 1β血浆水平 (5 6 pg/ml± 15 pg/ml)仍明显高于再灌注前 (2 8pg/ml± 9pg/ml,P <0 0 1)。 (2 )兔AMI对照组与rhIL 1ra各组之间心肌缺血面积差异均无显著意义 ,但各rhIL 1ra组梗死心肌面积均小于对照组 ,组间差异呈剂量依赖性 (47%±7% ,34%± 8% ,31%± 6 %比 6 1%± 11% ,分别P <0     

Cytokine production from peripheral blood mononuclear cells in patients with ankylosing spondylitis and their first-degree relatives

BACKGROUND: To understand the cytokine levels in different disease activities of patients with ankylosing spondylitis (AS), we measured proinflammatory and antiinflammatory cytokine production from peripheral blood mononuclear cells (PBMC) in patients with AS and their first-degree relatives (FDR). METHODS: PBMC were obtained from 26 patients with AS and 24 FDR and then stimulated with PHA for 72 h. In the supernatants, the following three cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), and IL-10, were measured by ELISA. Disease activity in AS patients was divided into high disease activity (Group 1) and low disease activity (Group 2), based on the Bath AS Disease Activity Index (BASDAI > or =4 or <4). Healthy FDR of AS patients (Group 3) and healthy subjects (Group 4) were used as a control group. RESULTS: TNF-alpha production from PBMC was significantly increased in Group 1 patients compared to Group 2 patients (1371 +/- 1008 pg/mL vs. 355 +/- 89 pg/mL, p <0.05) or FDR (1371 +/- 1008 pg/mL vs. 552 +/- 89 pg/mL, p <0.05) or healthy subjects (1371 +/- 1008 pg/mL vs. 436 +/- 114 pg/mL, p <0.01). IL-1beta also showed a similarly significant difference between the two groups (Group 1 vs. Group 2, Group 1 vs. Group 4) (p <0.05). In contrast, IL-10 was significantly decreased in Group 1 when compared to Group 2 (126 +/- 64 pg/mL vs. 272 +/- 150 pg/mL, p <0.05). CONCLUSIONS: Patients with high BASDAI had increased production of TNF-alpha and IL-1beta compared to those with low BASDAI or healthy FDR, suggesting that proinflammatory cytokines may play an important role during active inflammation.     

Chronic treatment of enbrel in rats with isoproterenol- induced congestive heart failure limits left ventricular dysfunction and remodeling

Objective To investigate the effect of chronic treatment of enbrel (EB), a TNF- α antagonist, in a well defined congestive heart failure (CHF) rat model and test the hypothesis that chronic treatment of EB in CHF rats may limit the progression of Left ventricular (LV) dysfunction and structure remodeling and decrease cardiac IL- 1β levels. Methods We measured cardiac conformation, contractile performance and cytokines level in 8 age- matched normal adult rats (control group) and 8 rats with isoproterenol (ISO)- induced Heart failure (ISO group) and 8 rats with ISO- induced lesion but received EB treatment (EB group). Results LV end diastolic diameter and LV end systolic diameter in EB group were significantly less and LV fractional shortening was significantly larger than ISO group (9. 2±0. 3 mm vs 9. 5±0. 2 mm, 5. 8±0. 5 mm vs 6. 5±0. 3 mm, 0. 37±0. 03 vs 0. 31±0. 02,P&lt;0. 05,P&lt;0. 01,P&lt;0. 01 respectively）, but there was no significant difference of LV posterior wall thickness at end diastole between the two groups; LV end systolic pressure (P ES ), dp/dt max in EB group were significantly greater than ISO group (104. 8±4. 6 mm Hg vs 98. 4±4. 9 mm Hg, 8395±940 mm Hg/s vs 6898±612 mm Hg,P&lt;0. 05,P&lt;0. 01 respectively), and LV end diastolic pressure (P ED ), dp/dt min , time constant of LV relaxation were significantly lower than ISO group (3. 8±0. 6 mm Hg vs 7. 1±0. 8 mm Hg, -5963±475 mm Hg/s vs -5030±316 mm Hg/s, 15. 4±0. 8 ms vs 21. 3±1. 4 ms,P&lt;0. 01, respectively）. Although cardiac contractile performance in the EB group was greatly improved, there still was a big gap when compared with the control group. The ratio of LV weight to body weight in the EB group was significantly higher than control group （2. 82±0. 07 mg/g vs 2. 28±0. 08 mg/g,P&lt;0. 01）, but there was no significant difference when compared with the ISO group. There was no significant difference between the serum level of TNF- α in EB group and ISO group, the it could not be detected in control group. TNF- α levels in LV of EB group was significantly higher than control group, （757. 6±46. 8 pg/g vs 367. 5±22. 7 pg/g,P&lt;0. 01）, but there was no significant difference when compared with ISO group. The IL- 1β level in LV of EB group was significantly lower than ISO group （356. 2±28. 5 pg/g vs 518. 4±32. 5 pg/g,P&lt;0. 05）, and it could not be detected in control group. The serum level of IL- 1β could not be detected in any rats. Conclusion EB administered as soon as possible when ISO induced myocardial necrosis occurs can greatly improve cardiac contraction, and the improvement may be partly due to a decrease in the IL- 1β level in LV, besides the direct blocking effect of EB on TNF- α. EB can alleviate cardiac remodeling by its effect on LVEDD.      

龈沟液微量样本中多种成分的检测

目的:以龈沟液(gingival crevicular fluid,GCF)样本为例,探寻微量样本中多种成分的检测方法.方法:根据酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)的基本原理,确认待检测成分之间无交叉反应,样本与包被抗体在4 ℃下进行低温反应,重复使用同一样本.39份龈沟液来自4例轻、中度牙周炎患者,龈沟液洗提液分为两份,一份样本纳入A组,用ELISA方法先检测白细胞介素6(interleukin-6,IL-6),再用此样本检测白细胞介素1β(interleukin-1β,IL-1β);另一份样本纳入B组,直接进行IL-1β检测.比较A组与B组IL-1β的检测结果.65份龈沟液样本选自9例来北京大学口腔医院修复科就诊患者,龈沟液洗提液分为两份,一份样本纳入C组,用ELISA方法先检测肿瘤坏死因子(tumor necrosis factor-α,TNF-α)后,再用此样本检测弹性蛋白酶;另一份纳入B组,直接进行弹性蛋白酶检测.比较A组与B组弹性蛋白酶的检测结果.结果:A和B组IL-1β光密度值分别为(0.5±0.4)和(0.5±0.4),差异无统计学意义(P=0.136),且高度相关(r=0.993,P=0.000);C和D组弹性蛋白酶吸光度值分别为(1.1±0.6)和(1.1±0.6),差异无统计学意义(P=0.943),且高度相关(r=0.979,P=0.000).结论:通过重复使用同一样本,可以达到用微量样本检测多种成分的目的.     

风心病瓣膜置换术患者体外循环围术期细胞因子的释放

To examine cytokines release related to cardiopulmonary bypass(CPB), the anthers investigated the possible differences in cytokines responses between patients undergoing prosthetic valve replacement (study group, n = 10) and those receiving closure of patent ductus arteriosus or pericardiectomy(control group, n = 9). Venous levels of interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interleukin-10 (IL-10) were measured at multiple time points before, during and after operation. As compared with pre-operative values, IL-2 levels in both groups decreased significantly (P < 0.05), and the levels of sIL-2R, IL-6, TNF-alpha, and IL-10 in both groups increased significantly at multiple time points post-operative (P < 0.01). All the values of cytokines returned to pre-operative levels at 7th post-operative day. Although there were no pre-operative differences in these cytokines between the two groups (P > 0.05), the post-operative changes of cytokines in study group was more obvious than that in control group (P < 0.01). In study group IL-10 rose to a peak value of around 620 pg/ml at the end of CPB while IL-6 and TNF-alpha levels reached their peak values of around 88 pg/ml and 52 pg/ml respectively at 1st post-operative day. IL-10 has been reported as an antiinflammatory cytokine. The preceding IL-10 peak value, as compared with the peak values of IL-6 and TNF-alpha, could be associated with the interplay and regulation of cytokine network. On the other hand, in control group the levels of IL-6, TNF-alpha, and IL-10 reached their peak values at 1st post-operative day simultaneously. The values were 34 pg/ml, 36 pg/ml, and 162 pg/ml respectively. This result suggests that besides surgical stress mediated cytokines production or suppression, CPB itself also results in obvious changes of cytokine metabolism. However further studies are needed to elucidate the underlying mechanisms and clinical value of post-operative cytokines production or suppression related to CPB in patients with prosthetic valve replacement.     

异丙肾上腺素对高血压患者单核细胞分泌白细胞介素6及肿瘤坏死因子α的影响

目的:观察异丙肾上腺素(isoproterenol, ISO)对高血压患者单核细胞分泌肿瘤坏死因子α(tumor necrosis factor-alpha ,TNF-α)及白细胞介素6(interleukin-6, IL-6)的影响.方法:选取17例健康志愿者和6例原发性高血压病患者(Ⅰ期)的静脉血样,分离得到单核细胞,用脂多糖(lipopolysaccharide,LPS)刺激细胞后,检测在有或无ISO存在条件下细胞上清液中TNF-α和IL-6含量.结果:(1)在LPS刺激下,高血压组单核细胞分泌TNF-α量较健康对照组显著增加[(1 897±393) ng/L vs. (975±473) ng/L, P＜0.01],但IL-6的分泌在两组间的差异无统计学意义[(5 532±796) ng/L vs. (6 092±2 249) ng/L];(2)在健康对照组和高血压组中,ISO能剂量依赖性抑制LPS引起的单核细胞TNF-α分泌,但对IL-6的分泌则无显著影响;(3)在Ⅰ期高血压患者的单核细胞中,ISO对TNF-α产生的抑制作用与健康对照组相比,差异无统计学意义(P》0.05),给予β肾上腺素受体拮抗剂普萘洛尔可拮抗这种效应.结论:β肾上腺素受体激动可抑制LPS引起单核细胞分泌TNF-α,而对IL-6的分泌无影响,这种抑制效应在Ⅰ期高血压病患者与健康志愿者之间差异无统计学意义.     

甲氨蝶呤对类风湿关节炎患者的治疗作用及对相关细胞因子的影响

目的: 研究甲氨蝶呤(methotrexate,MTX)对活动性类风湿关节炎(rheumatoid arthritis,RA)患者的疗效,探讨该药治疗类风湿关节炎的免疫学机制,了解MTX对RA患者血清中Th1/Th2型细胞因子的作用,为临床MTX治疗RA提供理论依据.方法:入选30例活动性RA患者,每周1次口服MTX(首次7.5 mg, 每周递增2.5 mg至15 mg ),疗程24周.采用美国风湿病学会(ACR)疗效评定标准,采用酶联免疫吸附双抗夹心法(ELISA)检测健康对照组以及RA患者MTX治疗前后血清中的细胞介素IL-1β,IL-6,IL-10,肿瘤坏死因子TNF-α和INF-γ浓度.结果: (1)MTX治疗第2周ACR20达到23%(7/30), 至24周时ACR20上升至70%(21/30),ACR70也升至10%(3/30).临床病情活动性指标(如晨僵、红细胞沉降率、压痛关节数、肿胀关节数、休息痛、患者及医生评价)较治疗前均有显著改善.(2) RA患者血清中的炎性细胞因子IL-6(46.83±35.81 vs. 20.92±17.98,P=0.028),TNF-α(162.52±107.63 vs. 18.32±14.36,P=0.001),INF-γ(67.79±43.76 vs. 35.78±27.51,P=0.004)等的水平均高于健康对照组,且差异具有统计学意义.抗炎性细胞因子IL-10的水平低于健康对照组(46.17±26.70 vs. 47.21±28.94),但差异无统计学意义(P=0.887).(3) MTX治疗RA后血清中Th1型细胞因子与治疗前相比,IL-1β(7.47±7.33,P=0.265),IL-6(26.01±25.64,P=0.025),INF-γ(41.53±13.49,P=0.015),TNF-α(123.36±89.61, P=0.018)等水平均明显降低,而Th2型细胞因子IL-10的水平明显提高(71.76±41.01,P=0.02).结论:MTX治疗活动性RA起效早,疗效显著.并且MTX可以下调RA患者血清中IL-1β,IL-6,TNF-α和INF-γ水平,同时上调IL-10水平,抑制了Th1型细胞因子的炎症作用,增强了Th2型细胞因子的效应,从而抑制或控制了RA的病情发展.     

类风湿关节炎滑膜细胞中白三烯B4诱导肿瘤坏死因子-α和白细胞介素-1βmRNA表达的测定

目的:探讨类风湿关节炎(rheumatoid arthritis, RA)滑膜细胞中白三烯B4(LTB4)诱导肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)mRNA表达的定量测定方法.方法:RA患者的滑膜细胞进行原代培养,加入外源性LTB4或者在LIT存在的情况下,分别加入MK-886(5-脂氧合酶激动蛋白抑制剂)和苯丁抑制素(Bestatin,LTA4水解酶抑制剂),采用TaqMan PCR来定量检测滑膜细胞中TNF-α和IL-1β mRNA水平的表达.结果:原代培养的RA滑膜细胞的基本TNF-α和IL-1β mRNA表达水平(TNF-α/GAPDH和IL-1β/GAPDH),分别为0.02±0.00和0.16±0.01.当加入外源性的LTB4 10^-9和10^-8 mol/L后,LTB4 10^-9 mol/L使TNF-α RNA水平的表达增高了7倍,LTB4 10^-8 mol/L使TNF-α mRNA水平的表达增高了15倍,LTB4 10^-8 mol/L使IL-1β mRNA水平增高了1倍.而加入LIT刺激内源性的LTB4增高后,使TNF-α和IL-1β mRNA水平分别增高了145倍和12倍.在LIT存在的情况下,加入LTB4合成抑制剂MK-886(1 μmol/L,10 μmol/L)后,使TNF-α mRNA水平的表达分别下降15%和66%,IL-1βm RNA水平的表达分别下降了41%和71% . 1100 mg/L 苯丁抑制剂使TNF-α和IL-1β mRNA水平表达分别降低了86%和79%.结论:RA滑膜细胞中LTB4可诱导TNF-α和IL-1β mRNA水平的表达,并有助于定量测定mRNA表达水平.     

A study on the serum levels of interleukin-1beta in bronchial asthma

Mononuclear phagocytes can be activated through an immunoglobulin E (IgE)- specific mechanism to release pro-inflammatory cytokine like interleukin-1beta (IL-1beta). The present study was conducted to show the inter-relationship between these two parameters in the serum of asthmatic patients. The study included 30 patients of asthma and 10 as control. Out of these 30 cases, 20 patients had stable and 10 had acute asthma. Of the 20 stable patients, 9 were allergic and 11 were non-allergic to either of the 12 allergens used for skin prick test. Serum IgE and IL-1beta levels were measured by enzyme linked immunosorbent assay (ELISA). Total serum IgE levels increased significantly (p < 0.05) in asthma [200.5 +/- 30.91 IU/ml, mean +/- standard error of mean (SEM)] in comparison with the controls (18.15 +/- 4.35 IU/ml). Serum IL-1beta level was higher in allergic (1.94 +/- 0.63 pg/ml) than in non-allergic patients (0.64 +/- 0.21 pg/ml) but it was not statistically significant (p > 0.05). The study suggests involvement of IgE and IL-1beta in the pathophysiology of allergic asthmatic condition. Further studies are required to delineate the inflammatory pathway in asthma and determine stages at which therapeutic interventions can be done.     

高迁移率族蛋白1诱导内皮细胞释放细胞因子的作用及其与脂多糖对白细胞介素6释放的协同效应

目的观察高迁移率族蛋白1(HMGB1)对人脐静脉内皮细胞(HUVEC)释放细胞因子的影响及其对脂多糖(LPS)诱导细胞因子白细胞介素6(IL-6)表达的作用。方法用LiquiChip液相蛋白芯片系统检测重组HMGB1蛋白(15ng/ml)诱导HUVEC11种细胞因子/趋化因子的水平变化;检测不同浓度HMGB1(0~75ng/ml)刺激后不同时间点(0、1、3、6、12和24h)HUVEC分泌IL-6的水平以及HMGB1(15ng/ml)与LPS(10ng/ml)共同刺激对HUVEC分泌IL-6的影响。结果HMGB1刺激后HUVEC分泌粒细胞-巨噬细胞集落刺激因子(GM-CSF)、干扰素γ(IFN-γ)、IL-6、IL-8和单核细胞趋化蛋白1(MCP-1)的水平明显升高(均P<0.01),分别是对照(未加刺激)的5.7、4.2、27.8、12.8和5.4倍;HMGB1蛋白以时间和剂量依赖方式诱导IL-6的分泌,在刺激后3~6h,IL-6水平开始增加,在6h时IL-6由对照的32pg/ml±21pg/ml增加到75pg/ml±22pg/ml(P<0.01),12h(453pg/ml±78pg/ml)~24h(901pg/ml±184pg/ml)持续升高(P<0.01);随着HMGB1浓度的增加,IL-6的水平也明显增加,当HMGB1浓度为3、15、75ng/ml时,IL-6分别是155pg/ml±33pg/ml、901pg/ml±184pg/ml、1508pg/ml±378pg/ml,与基础值32pg/ml±21pg/ml相比,差异有统计学意义(P<0.01)。分别用LPS(10ng/ml)和HMGB1(15ng/ml)单独刺激HUVEC时,IL-6的含量从基础的32pg/ml±22pg/ml分别增加至289pg/ml±42pg/ml和901pg/ml±184pg/ml(均P<0.01);如果用二者共同刺激HUVEC,IL-6的生成量大大增加(2361pg/ml±299pg/ml),二者存在协同作用(F=69.405,P<0.01)。结论HMGB1蛋白可诱导HUVEC释放多种炎性细胞因子;HMGB1诱导IL-6的上调具有时效性和量效性关系,并可协同LPS刺激HUVEC释放IL-6,在脓毒症的发生和发展中起重要作用。