骨髓间充质干细胞对烟雾吸入性损伤兔早期炎症因子分泌的影响

目的 探讨骨髓间充质干细胞(MSCs)对烟雾吸入性损伤早期外周血及肺组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-1β、IL-6、IL-10)分泌的影响.方法 全骨髓培养法体外培养兔MSCs,用流式细胞术鉴定.将56只健康新西兰大耳白兔按随机数字表法分为正常对照组(C组,n=8)、烟雾吸入性损伤组(S组,n=24)、烟雾吸入性损伤+MSCs移植组(M组,n=24),后两组再分为伤后2、4、6 h亚组,每组8只.采用酶联免疫吸附法(ELISA)检测血浆及肺组织匀浆液中促炎因子TNF-α、IL-1β、IL-6及抗炎因子IL-10的含量.结果 与C组比较,S组各时间点血浆促炎、抗炎因子均显著升高;各时间点肺组织促炎因子显著升高,抗炎因子无明显变化.与S组比较,M组各时间点血浆促炎因子显著下降,抗炎因子显著升高[6 h时TNF-α(μg/L):1.7±1.7比4.1±1.6,IL-1β(ng/L):9.9±1.7比21.2±2.6,IL-6(μg/L):1.0±0.3比1.3±0.2,IL-10(ng/L):15.2±4.4比7.9±3.5,均P＜0.05];各时间点肺组织促炎因子显著降低,而抗炎因子仅在4 h、6 h显著升高[6 h时TNF-α(ng/L):503.0±156.4比587.7±171.2,IL-1β(ng/L):0.4±0.2比0.6±0.2,IL-6(ng/L):155.2±13.7比350.2±20.3,IL-10(ng/L):23.3±5.4比11.0±5.6,均P＜0.05].结论 MSCs移植能降低烟雾吸入性损伤早期促炎因子水平,升高抗炎因子水平,改善全身炎症反应,对烟雾吸入性损伤肺组织具有一定的保护作用.

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Objective To explore the effect of bone marrow mesenchymal stem cells (MSCs) engraftment on secretion of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, IL-6, IL-10) in peripheral blood and lung homogenates in the early stages of smoke inhalation injury. Methods MSCs were proliferated by the method of whole marrow culture and identified by flow cytometry. Fifty-six healthy New Zealand rabbits were randomly divided into control group (C group, n=8), smoke inhalation injury group (S group, n=24)and smoke inhalation injury+MSCs engraftment group (M group, n=24). The latter two groups were subdivided into 2, 4, 6 hours after injury subgroups, with 8 rabbits in each group. The levels of TNF-α,IL-1β, IL-6 and IL-10 in peripheral blood and lung homogenates were measured by enzyme-linked immunosorbent assay (ELISA). Results Compared with C group, concent of pro-inflammatory and anti-inflammatory cytokines in peripheral blood at each time point in S group were increased significantly.The concent of pro-inflammatory cytokines in lung homogenate at each time point in S group was significantly higher than thoae in C group, and that of anti-inflammatory cytokines showed no significant changes.Compared with the S group, concent of pro-inflammatory cytokines in peripheral blood in M group was decreased significantly, and that of anti-inflammatory cytokines was increased significantly [6 hours TNF-α(μg/L):1.7±1.7 vs. 4.1±1.6, IL-1β (ng/L): 9.9±1.7 vs. 21.2±2.6, IL-6 (μg/L): 1.0±0.3 vs.1.3 ± 0. 2, IL-10 (ng/L): 15. 2 ± 4. 4 vs. 7. 9 ± 3.5, all P＜0.05]. Concent of pro-inflammatory cytokines at each time point in M group was decreased significantly when compared with S group in lung homogenate,while only anti-inflammatory cytokine at 4 hours and 6 hours was increased significantly [6 hours TNF-α (ng/L): 503. 0±156. 4 vs. 587.7±171.2, IL-1β (ng/L): 0.4±0.2 vs. 0.6±0.2, IL-6 (ng/L): 155.2±13.7 vs. 350.2±20.3, IL-10 (ng/L): 23.3±5.4 vs. 11.0±5.6, all P＜0.05]. Conclusion MSCs engraftment could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the early stages of smoke inhalation injury, thus amelioratea inflammatory reaponse, which confers protective effect on smoke inhalation injury.     

转基因阿尔茨海默病小鼠脑内炎症反应的诱发因素研究

目的 观察炎症反应在转基因阿尔茨海默病(AD)小鼠脑组织中的变化,探讨AD脑内炎症反应的诱发因素.方法 选用3、12个月龄转人β-淀粉样前体蛋白/早老素-1(APP/PS1)基因AD小鼠及正常野生型(WT)小鼠,分别应用免疫组织化学法和ELISA法观察脑内淀粉样斑块、炎性因子[白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)α、前列腺素(PGE)2]的变化.结果 3个月龄APP/PS1基因AD小鼠脑组织中无淀粉样斑块沉积,未发现激活的星型胶质细胞和小胶质细胞,炎性因子IL-1β、IL-6、TNFα、PGE2的含量与WT小鼠差异无统计学意义(P均＞0.05).12个月龄转APP/PS1基因AD小鼠脑组织中有大量淀粉样斑块沉积,并伴有大量激活的星型胶质细胞和小胶质细胞,炎性因子IL-1β、IL-6、TNFα、PGE2的含量[分别为(56.02±9.04)、(8.66±0.83)、(97.48±26.58)、(72.18±21.01)ng/g]较WT小鼠[分别为(29.81±6.03)、(7.73±0.74)、(61.98±11.11)、(37.23±10.96)ng/g]及3个月龄转APP/PS1基因AD小鼠[分别为(30.05±3.53)、(7.43±1.17)、(59.34±10.47)、(42.56±5.93)ng/g]显著增加(P＜0.05或P＜0.01).结论 在淀粉样斑块形成之前,转APP/PS1基因AD小鼠脑组织中无明确的炎症反应;而淀粉样斑块沉积之后,脑组织中有显著的炎症反应;AD脑内炎症反应与淀粉样斑块形成密切相关,淀粉样蛋白(Aβ)沉积是导致脑内炎症反应的直接诱发因素.

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Objective To observe the changes of cerebral inflammation-related markers in brain of a transgenic mouse model of Alzheimer's disease (AD) ,and to determine the causative factor to the development of cerebral inflammation in AD. Methods 3- and 12-month-old β-amyloid protein precursor ( APP)/presenilin (PSI) transgenic mice and age-matched wild-type mice (WT) were used in the study. The changes of amyloid plaques, inflammatory factors ( interleukin 1β ( IL-1β ); interleukin 6( IL-6 ); tumor necrosis factor α (TNFα) ;prostaglandin E2 (PGE2)) in the brains among these mice were measured by immunohistochemistry and ELISA. Results Immunohistochemical analysis revealed that no amyloid plaques and activated astrocytes as well as microglia were observed in the 3-month-old APP/PS1 mice. There were no significant differences in the levels of inflammatory factors (IL-1β, IL-6 ,TNFα,and PGE2) between the 3-month-old APP/PS1 and WT mice ( Ps ＞ 0. 05 ). However, abundant amyloid plaques accompanied by a remarkable increase of activated astrocytes and microglia were found in the brain of the 12-month-old APP/PS1 mice. The levels of inflammatory factors (IL-1β,IL-6,TNFα, and PGE2 ) were significantly increased in the 12-month-old APP/PS1 mice ([56. 02 ±9. 04] ng/g, [8. 66 ±0.83] ng/g, [97.48 ±26.58] ng/g, [72. 18 ±21.01] ng/g) than in the WT mice ([29. 18 ± 6. 03] ng/g, [7. 73 ± 0. 74] ng/g, [61.98 ±11.11] ng/g, [37. 23 ± 10. 96] ng/g) and the 3-month-old APP/PS1 mice ( [30. 05 ± 3.53] ng/g, [7.43 ± 1.17] ng/g, [59.34 ± 10. 07] ng/g, [42. 56 ±5.93] ng/g) (P＜0.05,or P＜0.01,respectively). Conclusion This study demonstrates that the APP/PS1mice did not show cerebral inflammation before the appearance of amyloid plaques, and exhibited remarkable inflammation after amyloid plaque deposition. These findings suggest that the induction of cerebral inflammation is tightly associated with amyloid plaque formation, and deposition of amyloid-beta protein (Aβ) may be the direct causative factor to the development of cerebral inflammation in AD.     

亚低温对急性肺损伤大鼠肺泡表面活性蛋白A含量的影响

目的 探讨亚低温对内毒素脂多糖(LPS)诱导急性肺损伤(ALI)大鼠肺泡表面活性蛋白A(SP-A)含量的影响.方法 按随机数字表法将40只雄性Wistar大鼠分组.采用气管内滴入LPS制备ALI动物模型;对照组气管内只滴人生理盐水.模型组和对照组分别于术后1 h和8 h处死8只大鼠;亚低温组于滴入LPS 1 h后将体温降低并维持在32.5～33.0℃,8 h后处死8只大鼠.各组分别于术前及术后1 h、8 h测定动脉血气,并计算氧合指数(PaO2/FiO2);采用酶联免疫吸附法检测支气管肺泡灌洗液(BALF)中SP-A含量;光镜下观察肺组织形态结构的变化.结果 气管内滴入LPS 1 h后,大鼠PaO2/FiO2均达到ALI的诊断标准.与对照1 h组比较,模型1 h组BALF中SP-A含量(μg/L)明显降低(53.27±1.95比74.81±6.55,P＜0.01);模型8 h组和亚低温8 h组SP-A含量(4.35±2.76和51.36±2.33)均较对照8 h组(70.81±5.01)明显降低,但亚低温8 h组SP-A含量较模型8 h组明显增高(均P＜0.01).光镜下观察,对照1 h和8 h组肺泡结构基本正常;模型8 h组肺组织炎症反应最重;模型1 h组和亚低温8 h组肺组织炎症反应较模型8 h组有所减轻.结论 亚低温能延缓内毒素诱导的ALI大鼠早期肺泡内SP-A含量下降的程度,在一定程度上可减轻肺损伤.

Abstract：

Objective To investigate the effect of hypothermia (HT) on the concentration of surfactant protein A (SP-A) during lipopolysaccharide (LPS) induced acute lung injury (ALI) in rats.Methods Forty male Wistar rats were randomly divided into three groups. The ALI model was reproduced by LPS intratracheal instillation; only saline was instilled intratracheally for control group. Rats in both model group and control group were sacrificed respectively at 1 hour and 8 hours (each n= 8). In HT group the body temperature was lowered to 32. 5 - 33.0 ℃ 1 hour after LPS instillation, and 8 rats were sacrificed st 8 hours. The arterial blood gas was determined in all the groups before and 1 hour and 8 hours after instillation of saline or LPS, and the oxygenation index (PaO2/FiO2) was calculated. The concentration of SP-A in bronchoalveolar lavage fluid (BALF) was determined by enzyme linked immunosorbent assay. The morphological changes in lung tissue of rats were observed under light microscope. Results At 1 hour after intratracheal instillation of LPS, the PaO2/FiO2 of each group reached the diagnostic criterion of ALI.Compared with control 1-hour group, the SP-A (μg/L) in BALF of model 1-hour group was decreased (53. 27±1.95 vs. 74. 81±6. 55, P＜0. 01); the SP-A in model 8-hour group and HT 8-hour group (4.35±2. 76 and 51.36±2. 33) was both obviously decreased compared with control 8-hour group (70. 81±-5. 01,both P＜0. 01). Compared with model 8-hour group, the SP-A of HT 8-hour group was obviously increased (P＜0. 01). Results of light microscopic examination, it was revealed that the alveolar structure of control 1-hour group and control 8-hour group was almost normal. Inflammatory response in lung tissues in model 8-hour group was found to be most serious; compared with model 8-hour group, inflammatory response in lung tissues in model 1-hour group and HT 8-hour group was reduced in certain degree. Conclusion A certain extent of HT may reduce lung injury of early endotoxin-induced ALI rats by delaying lowering of alveolar SP-A levels.     

慢性血吸虫感染对脓毒症小鼠保护作用的研究

目的 初步探讨慢性血吸虫(SJ)感染对脓毒症小鼠的保护作用及其机制.方法 选择BALB/c雄性小鼠,按随机数字表法分组进行三部分实验.实验1:经腹部皮肤接种SJ尾蚴感染8周建立慢性SJ感染模型,分为正常组和SJ组,每组10只;用酶联免疫吸附法(ELISA)检测血清白细胞介素(IL-4和IL-10)、肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)水平,实时荧光定量聚合酶链反应(PCR)检测腹腔巨噬细胞IL-10和TNF-α的mRNA表达,了解慢性SJ感染小鼠免疫状态.实验2:以脂多糖(LPS)腹腔注射诱导小鼠脓毒症模型,分为LPS组和SJ-LPS组,每组15只;用ELISA法动态观察注射LPS后0、24、48和72 h细胞因子的变化,0 h的水平相当于正常小鼠和SJ感染8周水平,观察慢性SJ感染对脓毒症过程的影响.实验3:分别以盲肠结扎穿孔术(CLP)和LPS诱导两种不同的脓毒症模型,评价慢性SJ感染对脓毒症小鼠72 h存活率的影响.结果 实验1:SJ组血清抗炎因子IL-4[(151.35±12.24)ng/L]和IL-10[(133.22±11.09)ng/L]水平较正常组[IL-4(56.32±8.66)ng/L,IL-10(48.17±7.23)ng/L]显著升高(均P＜0.05),并可使巨噬细胞向替代活化性巨噬细胞分化,慢性SJ感染使腹腔巨噬细胞高表达IL-10 mRNA(SJ组4.46±1.82,正常组1.52±0.60),抑制TNF-α mRNA表达(SJ组1.61±0.93,正常组2.32±1.03,均P＜0.05).实验2、3:慢性SJ感染小鼠血清IL-4、IL-10于注射LPS后0 h即显著升高,随后下降,至72 h仍明显高于LPS组[IL-4(ng/L):92.2±7.6比41.5±4.5;IL-10(ng/L):92.1±7.8比35.6±4.0,均P＜0.05];TNF-α、IFN-γ均于24 h达峰值后逐渐下降,至72 h SJ-LPS组仍显著低于LPS组[TNF-α(ng/L):82.9±5.6比91.5±5.2;IFN-γ(ng/L):44.1±4.8比52.6±4.0,均P＜0.05].慢性SJ感染可明显改善CLP或LPS所致脓毒症小鼠的存活率(CLP:80%比20%,LPS:70%比30%,均P＜0.05).结论 慢性SJ感染可使脓毒症小鼠血清抗炎因子升高,存活率上升,从而起到保护作用.

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Objective To preliminarily study the protective effect of chronic schistosoma japonica (SJ)infestation against sepsis in mice and its mechanism. Methods BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10), tumor necrosis factor-α(TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA).Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3 : two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. Results Experiment 1, compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L],chronic SJ infestation showed an increase in serum IL-4 [(151. 35 ± 12. 24) ng/L] and IL-10 [(133. 22 ±11. 09) ng/L, both P＜0. 05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4. 46±1. 82, normal group 1. 52±0. 60) and inhibited TNF-α mRNA expression (SJ group 1. 61±0.93, normal group 2. 32±1.03) in abdominal macrophages (both P＜0. 05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92. 2±7. 6 vs.41.5±4. 5; IL-10 (ng/L): 92. 1±7. 8 vs. 35. 6±4. 0, both P＜0. 05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPSgroup at 72 hours [TNF-α (ng/L): 82. 9±5. 6 vs. 91. 5±5. 2; IFN-γ (ng/L): 44.1±4. 8 vs. 52. 6±4. 0,both P＜0. 05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P＜0.05). Conclusion Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.     

高迁移率族蛋白B1在失血性休克致急性肺损伤中的作用

Objective To investigate the role of high mobility group box1 (HMGB1) in hemorrhagic shock-induced acute lung injury (ALI) in mouse model. Method Forty-two healthy male BALB/c mice were randomly divided into three groups: control group, hemorrhagic shock (HS) group and anti-HMGB1 treatmeut group. Car-diac puncture, was performed to induce ALI in mice. IL-β and TNF-α in lung tissues were detected by ELISA. Ex-pression of HMGB1 was determined by Westemblot. Myeloperoxidase (MPO) and Evens blue dye (EBD) in lung tissues were measured by colorimetry. Pathology of lung tissue was observed. ANOVA and LSD-t test were used in statistical analysis. Results Pulmonary blood vessel permeability increased 2 h after HS, pathology results re-vealed a diffused inflammatory cell infiltration in mouse lung tissue. Levels of IL-1β and TNF-α in lung tissues sig-nificantly increased at the early stage of ALI (4 h) compared to controls [IL-1β(333.83±31.18) vs. (284.83± 30.49), P =0.014; TNF-α(805.00±114.67) vs. (584.17±181.17), P =0.023]. Significant increase of HMGB1 in lung tissues occurred at the late stage of ALI (16～48 h) [HMGB1/β-actin (0.99±0.16) vs. (0.50 ±0.18), P =0.003]. The levels of MPO and EBD in ALl group significantly increased compared to controls, and reached the peak at4 h and 24 h respectively [MPO (38.33±3.88) vs. (8.00±0.89), P <0.01;EBD (20.05±2.79) vs. (4.63±0.43), P <0.01]. The peak level of MPO and EBD significantly decreased in anti-HMGB1 groups compared to that of ALI group [MPO (25.67±1.63) vs. (38.33±3.88), P < 0.01 ; EBD (5.68±0.53) vs. (20.05±2.79), P <0.001]. The infiltration of neutrophils and the destruction of the pul-monary cell walls ameliorated in anti-HMGB1 groups, especially in mice treated with anti-HMGB1 antihedy for 24 h. Conclusions HMGBI acts as a late pminfiammatory mediator in HS-induced ALI. Blockade of HMGB1 de-creases HS-induced pathophysiologic change of ALI in mice.     

普通肝素雾化吸入对内毒素性肺损伤大鼠肺泡局部凝血及炎症反应的影响

目的 观察内毒素诱导急性肺损伤(ALI)大鼠雾化吸入普通肝素(UFH)后的肺部局部效应,探讨其对肺泡内凝血、纤溶和炎症反应的作用.方法 87只雄性Wistar大鼠按随机数字表法分为假损伤组、模型组、肝素治疗组(HT组)和肝素预防组(HP组).静脉注射(静注)内毒素脂多糖(LPS)制备ALI模型.HP组和HT组分别于注射LPS前后给予UFH雾化吸入,模型组和假损伤组则雾化吸入生理盐水.各组分别于静注LPS后6、12和24 h处死大鼠进行肺泡灌洗,采用酶联免疫吸附法测定支气管肺泡灌洗液(BALF)中凝血酶-抗凝血酶复合物(TAT)、组织型纤溶酶原激活物(t-PA)、尿激酶型纤溶酶原激活物(u-PA)、纤溶酶原激活物抑制剂-1(PAI-1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平.结果 损伤后6 h,模型组BALF中TAT浓度(μg/L:3.346±0.585)最高,其次是HT组(2.764±0.100),HP组(2.564±0.216)最低(均P＜0.05);HP组t-PA(μg/L:3.037±0.524)最高,HT组(2.494±0.191)其次,模型组(1.716±0.125)最低(均P＜0.05);HP组u-PA(μg/L)高于模型组(0.411±0.118比0.303±0.049,P＜0.05);HP组PAI-1(μg/L)明显低于HT组和模型组(2.296±0.246比2.597±0.425、2.834±0.198,均P＜0.05),直至12 h时HP组仍低于HT组(1.273±0.441比1.817±0.252,P＜0.05);HT组和HP组TNF-α(ng/L)显著低于模型组(68.154±3.915、36.990±6.539比77.001±4.485),且HP组低于HT组(均P＜0.05),至12 h HP组(15.287±4.754)仍最低,与HT组和模型组(26.756±5.336、23.674±4.398)比较差异有统计学意义(均P＜0.05).模型组、HT组和HP组各时间点IL-6水平均无明显差异.结论 内毒素性ALI大鼠雾化吸入UFH后起到了抑制局部凝血、减弱纤溶抑制、促进纤溶、降低炎症反应的作用,预防性吸人UFH比治疗性吸入效果更显著,最佳效应时间在注射后6 h.

Abstract：

Objective To observe the local changes in alveoli in intravenous endotoxin-induced acute lung injury (ALI) rat model after inhalation of aerosolized unfractioned heparin(UFH), and to observe its effects on coagulability, fibrinolysis and inflammatory response. Methods Eighty-seven male Wistar rats were divided into groups according to table of random number: sham, model, heparin therapy (HT) and heparin prophylaxis(HP). Endotoxin-induced ALI model was reproduced by intravenous administration of lipopolysaccharide (LPS). Rats in Hp and HT groups received aerosolized UFH before and after injection with LPS respectively, while rats in both sham and model groups inhaled aerosolized normal saline. Rats in each group were respectively sacrificed at 6, 12 and 24 hours after intravenous administration of LPS, and bronchoalveolar lavage fluid (BALF) was collected. Enzyme-linked immunosorbent assay was used to measure the level of thrombin-antithrombin (TAT), tissue-type plasminogen activator (t-PA),urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor- (TNF-α), interleukin-6 (IL-6) in BALF. Results At.6 hours after injury, the level of TAT (μg/L) in model group (3.346±0. 585) was highest, that in HT group (22. 764±0. 100) was higher, and that in HP group (2. 564±0. 2216) was lowest in BALF (all P＜0. 05). The t-PA(μg/L) concentration in HP group (3.037±0. 5224) was highest, that in HT group (22. 494±0. 191) was higher, and that in model group (1. 716±0. 1225) was lowest (all P＜0. 05). Compared with model group, u-PA (μg/L) level in HP group dramatically enhanced (0. 411±0. 118 vs. 0. 303±0. 049, P＜0. 05). The concentration of PAI-1 (μg/L) in HP group was significantly lower than that of HT and model groups (22. 2296 ± 0. 2246 vs. 22.597±0. 4225,2.834±0.198, both P＜0. 05). In HP group, it was still lower than that in HT group at 12 hours (1.2273±0. 441 vs. 1. 817±0. 252, P＜0. 05). TNF-α(ng/L) levels in HT and HP groups were markedly lower compared with model group (68. 154±3. 915, 36. 990±6. 539 vs. 77. 001±4. 485) at 6 hours, and the level in HP group was lower than that in HT group (all P＜0. 05). TNF-α concentration in HP group was still the lowest at 122 hours (15.2287±4. 754), and there was significant difference compared with HT and model groups (26. 756±5. 336, 23. 674±4. 398, both P＜0. 05). The levels of IL-6 were not distinctively different among model, HT and HP groups at various time-points. Conclusion It was proved that inhalation of aerosolized UFH resulted in lowering local coagulability, alleviating fibrinolytic depression, improving fibrinolysis, and attenuating inflammation in endotoxin-induced ALI rat model. More prominent results will be obtained when it was use as a prophylactic measure. The optimal time of usage is 6 hours after endotoxin injection.     

白细胞介素-10和肿瘤坏死因子-α与高血压肾损害的相关性研究

目的 探讨肿瘤坏死因子-α(TNF-α)和白细胞介素-10(IL-10)在高血压肾损害患者中的变化及其相关性.方法 将73例原发性高血压患者(原发性高血压组)根据其尿蛋白排泄率不同又分为2个亚组:单纯高血压组37例,高血压肾损害组36例;采用放射免疫法检测血清TNF-α、IL-10浓度.同期选择30名健康体检者作为正常对照组.结果 原发性高血压组TNF-α高于正常对照组[(2.91±0.94)μg/L与(0.98±0.35)μg/L,P＜0.01],其中高血压肾损害组TNF-α高于单纯高血压组[(3.75±0.88)μg/L与(1.87±0.58)μg/L,P＜0.01].原发性高血压组IL-10低于正常对照组[(19.2±5.8)μg/L与(28.6±5.7)μg/L,P＜0.01],其中高血压肾损害组IL-10又低于单纯高血压组[(15.4±4.3)μg/L与(22.5±5.9)μg/L,P＜0.01].原发性高血压组TNF-α、IL-10与尿蛋白排泄率有相关性(r=0.703,P＜0.001 ;r=-0.613,P＜0.001),而与血压水平无相关性.结论 高血压肾损害患者TNF-α升高,IL-10水平降低,细胞因子系统的失衡可能参与了高血压肾损害的进展.

Abstract：

Objective To investigate the changes of the serum levels of necrosis alpha (TNF-o)and interleukin 10( IL-10 )in patients with hypertensive renal damage,and to study the correlation of TNF-α and IL-10 with the hypertensive renal damage. Methods Seventy three patients with primary hypertension were divided into two groups according to their urinary albumin excretion rate(UAER): simple hypertensive group( n = 37 ),hypertensive renal damage group(n =36). TNF-α and IL-10 were measured using radioimmune assay. Thirty normotensive healthy persons were selected as normotensive control group. Results TNF-α were significantly higher and IL-10 significantly lower in patients with essential hypertension than those in normotensive control group(TNF-α: [2.91 ±0.94]μg/L vs [0.98 ±0.35]μg/L,P＜0. 05;IL-10:[ 19.2 ±5.8]μg/L vs [28.6±5. 7] μg/L,P ＜0. 01 ) ,and in patients with hypertension,those with renal damage had higher TNF-α and lower IL-10 than those without( TNF-α: [ 3.75 ± 0. 88 ] μg/L vs [ 1.87 ± 0. 58 ] μg/L, P ＜ 0. 01; IL-10: [ 15. 4 ± 4. 3 ]μg/L vs [ 22. 5 ± 5.9 ] μg/L, P ＜ 0. 01 ), with statistically significant difference between groups ( P ＜ 0. 01 ).TN F-α and IL- 10 were found to have correlations with UAER ( r = 0. 703, P ＜ 0. 001; r = - 0. 613, P ＜ 0. 001 ),but no correlation with the level of blood pressure. Conclusion TNF-α increased and IL-10 decreased significantly in patients with hypertensive renal damage, which indicates that the imbalanced cytokine network may play a role in the pathological mechanisms of hypertensive renal damage.     

谷氨酰胺对内毒素血症大鼠肠道损伤的保护作用及对血红素加氧酶-1表达的影响

目的 探讨谷氨酰胺(Glu)对内毒素血症大鼠肠道损伤的保护作用以及对血红素加氧酶-1(HO-1)表达的影响.方法 按随机数字表法将32只雄性SD 大鼠分为正常对照组、模型组、Glu组和Glu+锌原卟啉(ZnPP)组,每组8只.腹腔注射内毒素脂多糖(LPS)10 mg/kg制备内毒素血症动物模型.Glu组注射LPS前12 h灌胃Glu 1 g/kg;Glu+ZnPP组注射LPS前12 h灌胃Glu 1 g/kg,注射LPS前1 h静脉注射ZnPP 10 mmol/kg.术后12 h取回肠组织,测定髓过氧化物酶(MPO)活性及肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)含量,并进行肠组织学评分;用免疫组化法检测回肠组织HO-1表达.结果 与正常对照组比较,模型组回肠组织学评分、MPO活性及TNF-α、IL-10含量显著升高[组织学评分(分):3.3±0.4比1.1±0.6,MPO活性(U/g):0.40±0.08比0.26±0.07,TNF-α含量(ng/g):25.2±6.9比6.5±2.8,IL-10含量(ng/g):27.6±10.2比5.7±2.9,均P<0.01];HO-1表达较低.与模型组比较,Glu组组织学评分、MPO活性和TNF-α含量明显减低[组织学评分(分):1.6±0.5比3.3±0.4,MPO活性(U/g):0.25±0.05比0.40±0.08,TNF-α含量(ng/g):13.4±3.2比25.2±6.9,均P<0.01],IL-10含量(ng/g)显著升高(47.3±5.5比27.6±10.2,P<0.01),HO-1表达明显增加.Glu+ZnPP组与模型组各指标比较差异无统计学意义.结论 Glu能明显增强内毒素血症大鼠肠组织HO-1表达,明显减轻肠道炎症反应,从而保护肠黏膜.

Abstract：

Objective To investigate the protective effect of glutamine (Glu) pretreatment on intestinal injury induced by endotoxin and expression of heme oxygenase-1 (HO-1) in rats.Methods Thirty-two male Sprague-Dawley (SD) rats were randomly divided into four groups (n=8 in each group): normal control group, model group, Glu group and Glu+zinc protoporphyrin (ZnPP) group. In model group, endotoxemia was produced by intraperitoneal injection of lipopolysaccharide (LPS, 10 mg/kg). In Glu group, the rats received intragastraiclly 1 g/kg of Glu 12 hours before LPS intraperitoneal injection. In Glu+ZnPP group, the rats received 1 g/kg of Glu by gavage 12 hours before LPS intraperitoneal injection and ZnPP 10 mmol/kg intravenously via tail vein 1 hour before LPS injection. The distal ileum was harvested in full thickness 12 hours after LPS injection. The myeloperoxidase (MPO) activity, tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in the intestine were determined, the pathologic changes were observed and expressed in Chiu grade. The expression of HO-1 was evaluated by immunohistochemistry method. Results Compared with normal control group, the Chiu grade, MPO activity, the content of TNF-α and IL-10 were significantly increased in model group [Chiu grade: 3.3±0.4 vs. 1.1±0.6, MPO activity (U/g): 0.40±0.08 vs. 0.26±0.07, TNF-α (ng/g): 25.2±6.9 vs. 6.5±2.8, IL-10 (ng/g): 27.6±10.2 vs. 5.7±2.9, all P<0.01], and the expression of HO-1 was decreased. Compared with model group, the Chiu grade, MPO activity, the content of TNF-α in Glu group were significantly decreased [Chiu grade: 1.6±0.5 vs. 3.3±0.4, MPO activity (U/g): 0.25±0.05 vs. 0.40±0.08, the content of TNF-α (ng/g): 13.4±3.2 vs. 25.2±6.9, all P<0.01], while the level of IL-10 (ng/g) elevated (47.3±5.5 vs. 27.6±10.2, P<0.01), and the expression of HO-1 was increased. There was no difference in above mentioned indexes between model group and Glu+ZnPP group. Conclusion Glu pretreatment significantly ameliorates the expression of HO-1 of intestinal tissue induced by LPS in rats, and intestinal mucosa is protected with alleviation of inflammatory reaction in intestinal tract.     

The Sonic hedgehog signaling pathway involved in the expression of RACK1 in the pulmonary microvascular endothelial cells induced by lipopolysaccharide北大核心CSCD

Objective To explore the effect of expression of protein kinase C receptor l(RACKl) induced by lipopolysaccharide (LPS) on Sonic hedgehog(SHH) signaling pathway in rat puhnonary microvascular endothelial cells (RPMVEC). Methods The healthy male SPF grade SD rat with 100-120 g body weight were gotten from the laboratory animal center of Annui province. Using immunocytochemistry method, the expression of RACK 1 protein in RPMVECs was detected, cultured RPMVECs were randomly divided into different groups as LPS dose-dependent group, SAG(smoothened Agonist, a SHH signaling pathway specific agonist) dose-dependent group, LPS time-dependent group, SAG time-dependent group and LPS+SAG group. In LPS dose-dependent groups, RPMVECs were cultured with 0.1, 1, 10 mg/L LPS for 8 h. In LPS time-dependent groups, RPMVECs were cultured with 10 mg/L LPS for 0, 2, 4, 8, 12, 24 h. In SAG dose-dependent groups, RPMVECs were cultured with 0.1, 1, 10 u, mol/L for 8 h. In SAG time-dependent groups, RPMVECs were cultured with 1 u, mol/L SAG for 0, 2, 4, 8, 12, 24 h. In LPS+SAG group, RPMVECs were cultured with 1 u. mol/L SAG 8 h after 10 mg/L LPS treatment for 1 h. In addition, blank group, LPS group and SAG group were set for control. Western blot were used to detect the level of RACK1 and RT-PCR were used to detect the expression of GLI-1 mRNA after intervention. Results Immunocytochemistry revealed that RACK1 were present in RPMVEC. 1. In LPS dose-dependent groups (0, 0.1, 1, 10 mg/L), the level of RACK 1 elevated as LPS dose increased correspondingly with inter-group difference (P<0.05); the relative expression levels of GLI-1 mRNA were (1.109 ± 0.063), (1.039 ± 0.135), (0.813 ± 0.066), (0.770 ± 0.105), (1 mg/L vs. 10 mg/L, P>0.05; the rest P<0.05). In LPS time-dependent groups, the relative expression level of RACK1 at 2 h (0.370 ± 0.010) was higher than that at 0 h (0.329 ± 0.008), peaked at 12 h (1.296 ± 0.048), and compared with 0 h, there was significant differences (F=l 272.204, P<0.05). The relative expression level of GLI-1 mRNA was decreased at 2 h (0.929 ± 0.007), and compared with 0 h(1.089 ± 0.042), there was significant differences (F=306.609, /><0.05). 2. In SAG dose-dependent groups, there was no significant difference in level of RACK1 between groups(all P>0.05). The relative expression levels of GLI-1 mRNA were (1.109 ± 0.063), (1.169±0.052), (3.468 ±0.128), (3.434±0.054), (0 μ.mol/L vs. 0.1 μ.mol/L and l μmol/L vs. 10 μ.mol/L, P>0.05, the rest P<0.05). Among SAG time-dependent groups, there was no significant difference in levels of RACK1 protein(P>0.05). The relative expression level of GLI-1 mRNA increased at 2 h (3.027 ± 0.065), and compared with 0 h (2.651 ± 0.123), there was significant differences (F= 132.841, P<0.05). 3. In LPS+SAG intervention groups, the expression of RACK1 was lower than that in LPS group (0.831 ± 0.040 vs. 1.189 ± 0.149, P<0.05), and the expression of GLI-1 mRNA was higher than that in LPS group (2.720 ± 0.130 vs. 0.796 ± 0.082, P<0.05). Conclusions The LPS up-regulates the expression of RACK 1 in RPMVECs, and the activated SHH signaling pathway can down-regulate the expression of RACK 1 induced by LPS in RPMVECs. © 2018 Chinese Medical Association. All rights reserved.     

脂多糖致肺血管内巨噬细胞释放炎症因子变化的研究

目的探讨肺血管内巨噬细胞（PIM）在感染性急性肺损伤（ALI）发病中的作用。方法按照Morton法分离、培养猪PIM，予10mg／L脂多糖（LPS）刺激。酶联免疫吸附法（ELISA）测定肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和IL-8的含量。结果LPS刺激后，PIM释放TNF-α、IL-6和IL-8增多，峰值分别出现在刺激后的1、4和6h，与刺激前相比，差异均有显著性（P均〈0．01）。结论LPS刺激后，PIM分泌多种细胞因子，其中TNF—α升高最早，提示其在ALI发病早期起重要作用；而IL-6、IL-8升高较晚，且后者持续时间较长，可能对ALI的病情进展起重要作用。细胞因子间的相互作用在ALI发病中似乎更为重要。     

高压氧对油酸诱导大鼠急性肺损伤作用的实验研究

目的 探讨高压氧(HBO)对油酸(OA)诱导大鼠急性肺损伤(ALI)的干预作用.方法 80只SD大鼠按随机数字表法分为4组.OA组30只,经鼠尾静脉注射OA 0.15 ml/kg制备ALI模型,分别于制模后4 h、3 d、7 d各随机活杀10只;OA+HBO组20只,在HBO治疗箱给予2.5 atm(1 atm=101.325 kPa)下单次治疗90 min,分别于HBO治疗后3 d、7 d各随机活杀10只;单纯HBO干预组20只,分别于HBO治疗后3 d、7 d各随机活杀10只;另设正常对照组10只.取腹主动脉血进行血气分析,测定血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6;取左肺标本,观察大体形态改变及镜下病理学改变;取右肺,测定湿/干重(W/D)比值.结果 OA组4 h后动脉血氧分压(PaO2,mm Hg,1 mm Hg=0.133 kPa)由107.70±5.37降至57.40±2.63;肉眼可见肺脏明显淤血、水肿;光镜下肺泡正常结构消失,间质水肿增宽,大量炎性细胞浸润,毛细血管明显扩张、透明膜形成;W/D比值较正常对照组明显增加(6.94±0.44比4.59±0.44,P<0.05),血清TNF-α、IL-1β、IL-6水平升高[TNF-α(μg/L):18.52±1.20比5.27±0.61,IL-1β(μg/L):13.73±1.37比6.13±1.51,IL-6(μg/L):14.51±1.21比11.14±0.89].经HBO治疗3 d、7 d时PaO2(mm Hg,3 d:79.20±1.68比59.00±2.70,7 d:94.30±3.77比74.00±3.85)、肺W/D比值(3 d:7.43±0.73比9.82±0.99,7 d:6.75±1.14比8.77±1.60)均较OA组同期有不同程度改善(P<0.05或P<0.01).治疗3 d后HBO有降低血清中IL-1β(μg/L)的作用(6.46±1.99比9.09±1.09,P<0.05).结论 HBO治疗有改善ALI大鼠氧合,促进肺水吸收、抑制部分炎症介质产生的作用.     

硫化氢对急性肺损伤大鼠白细胞介素的调节

目的 研究气体信号分子硫化氢(H2S)在油酸诱导的大鼠急性肺损伤(ALI)中的作用及其在ALI中对炎症反应的调节.方法 采用鼠尾静脉缓慢注射油酸(OA,0.01 ml/kg)复制大鼠急性肺损伤模型,将SD大鼠49只随机分为三组:对照组、OA组及OA+硫氢化钠(NaHS)组.OA组和OA+NaHS组分为2 h、4 h、6 h三个观察点.对照组鼠尾静脉注射生理盐水(0.01 ml/kg),观察6 h.测定动脉血氧分压(PaO2)、肺湿干重比(W/D)、肺泡灌洗液中性粒细胞百分比(PMN%),光镜下半定量肺损伤评分(IQA),用敏感硫电极法检测血浆、肺组织匀浆H2S含量,用双抗体夹心ELISA法测定血浆、肺组织匀浆IL-6、IL-8和IL-10的含量.多组样本的均数的比较采用单因素差分析.结果 油酸鼠尾静脉注射可引起肺组织明显的形态学改变;与对照组比,OA组可见PaO2下降,W/D、PMN%、IQA增加(P＜0.01),在2 h、4 h、6 h时,血浆中H2S[对照:(36.58±6.80)μmol/L,2 h:(21.30±2.75)μmol/L,4 h:(20.63±1.26)μmol/L,6 h:(20.00±1.60)μmol/L,P＜0.01]以及肺匀浆中H2S[对照:(27.61±2.20)μmol/L,2 h:(20.67±1.37)μmol/L,4 h:(20.79±1.10)μmol/L,6 h:(18.92±0.75)μmol/L,P=0.000]均明显降低,血浆中IL-6[对照:(73.95±14.68)pg/ml,2 h:(186.70±23.85)pg/ml,4 h:(238.50±26.46)pg/ml,6 h:(215.95±25.86)pg/ml,P＜0.01]、肺匀浆中IL-6[对照:(60.58±12.91)pg/ml,2 h:(160.32±24.57)pg/ml,4 h:(195.27±46.28)pg/ml,6 h:(185.66±17.42)pg/ml,P＜0.01]均显著增加,血浆IL-8[对照:(80.69±20.42)pg/ml,2 h:(184.11±19.51)pg/ml,4 h:(286.20±53.34)pg/ml,6 h:(241.30±45.85)pg/ml,P＜0.01]、肺匀浆中IL-8[对照:(69.14±15.96)pg/ml,2 h:(174.10±20.36)pg/ml,4 h:(249.02±31.17)pg/ml,6 h:(237.74±34.18)pg/ml,P＜0.01]均显著增加,血浆IL-10[对照:(39.78±8.97)pg/ml,2 h:(111.18±11.46)pg/ml,4 h:(115.60±13.91)pg/ml,6 h: (102.41±9.93)pg/ml,P＜0.01]、肺匀浆中IL-10[对照:(71.86±14.19)pg/ml,2 h:(126.96±18.72)pg/ml,4 h:(151.88±27.61)pg/ml,6 h:(137.28±14.22)pg/ml,P＜0.01]含量均显著增加.而预先给予NaHS能减轻油酸引起的肺损伤,显著提高血浆和肺匀浆中H2S含量[4 h,血浆:(26.67±3.44)μmol/L vs(20.63±1.26)μmol/L,P=0.042;肺匀浆:(23.20±1.48)μmol/L vs(20.79±1.10)μmol/L,P=0.011;6 h,血浆:(26.98±4.93)μmol/L vs(20.00±1.60)μmol/L,P=0.022;肺匀浆:(21.43±1.79)μmol/L vs(18.92±0.75)μmol/L,P=0.016),同时血浆IL-6(185.37±21.98 pg/ml vs 238.50±26.46 pg/ml,4 h,P＜0.01;(124.22±21.84)pg/ml vs(215.95±25.86)pg/ml,6 h,P＜0.01 ]、IL-8[(199.40±34.56)pg/ml vs(286.20±53.34)pg/ml,4h,P＜0.01;(146.58±20.23)pg/ml vs(241.30±45.85)pg/ml,6 h,P＜0.01]含量明显降低,而IL-10含量增高[(154.48±18.08)pg/ml vs(115.60±13.91)pg/ml,4 h,P=0.000;(138.06±20.01)pg/ml vs(102.41±9.93)pg/ml,6 h,P=0.1300].结论 油酸诱导的大鼠ALI内源性H2S生成减少.外源性H2S通过抑制炎症因子IL-6和IL-8的表达,增加抗炎因子IL-10的表达,进而改变炎症/抗炎因子之间的比例,对油酸诱导的大鼠急性肺损伤起保护作用.     

脂多糖诱导血管平滑肌细胞分泌白细胞介素-6及p38丝裂素活化蛋白激酶的调控作用

目的 探讨p38丝裂素活化蛋白激酶(MAPK)信号转导通路在脂多糖(LPS)作用下血管平滑肌细胞(VSMC)分泌白细胞介素-6(IL-6)中的调节作用.方法 将体外培养的大鼠胸主动脉VSMC分为LPS刺激组、p38MAPK抑制剂SB203580干预组、SB203580对照组和溶液对照组.LPS组以终浓度100μg/L的LPS与VSMC共同孵育;干预组VSMC以p38MAPK抑制剂SB203580 10μmol/L预处理2 h,再加入终浓度100 9g/L的LPS共同孵育;对照组仅以SB203580 10 μmol/L预处理2 h;溶液组仅加入去血清培养液培养.各组于培养0、3、6、12、24 h后采用实时聚合酶链反应(real-time PCR)和酶联免疫吸附法(ELISA)分别检测细胞IL-6 mRNA和上清液中IL-6蛋白表达.结果 LPS刺激3 h,VSMC中IL-6 mRNA和蛋白表达即出现明显增高CmRNA(21.3±3.2)×104,蛋白(296.2±19.6)ng/L],12 h达高峰CmRNA(131.4±11.2)×104,蛋白(897.7±34.0)ng/L],24 h有所降低[mRNA(15.3±4.7)×104,蛋白(194.3±24.0)ng/L],但仍显著高于溶液组(mRNA(9.4±1.9)×104,蛋白(29.4±4.4)ng/L,均P<0.05].干预组3、6、12 h可明显抑制LPS诱导VSMC中IL-6的分泌[mRNA(15.4±3.6)×104、(43.2±6.6)X 104、(56.2±5.5)×104,蛋白(180.3±23.6)、(432.2±56.8)、(546.2±57.9)ng/L,均P<0.05].结论 LPS诱导VSMC可明显增加IL-6的mRNA和蛋白表达,p38MAPK抑制剂SB203580可显著抑制IL-6转录和蛋白合成,表明p38MAPK信号转导通路可能通过直接或间接作用参与了IL-6的分泌调节作用.     

神经降压素对内毒素诱导的急性肺损伤的保护作用及机制

目的探讨神经降压素在内毒素（LPS）所诱导的小鼠急性肺损伤（ALI）中的保护作用,并明确其在炎症反应中的作用。 方法100只健康无特异病原级性C57BL/6小鼠（雄性50只,雌性50只）,体质量（20~25 g）,由上海南方模式实验动物中心提供。所有小鼠随机分为以下5组：①正常对照组：60 μl无菌磷酸缓冲盐溶液（PBS）滴鼻处理小鼠;②急性肺损伤模型组：60 μg/只LPS滴鼻处理;③20 mg/kg神经降压素组：先使用LPS诱导肺损伤,诱导后1 h通过尾静脉注射的方式给予20 mg/kg神经降压素处理;④40 mg/kg神经降压素给药组：先使用LPS诱导肺损伤,诱导后1 h通过尾静脉注射的方式给予40 mg/kg神经降压素处理;⑤80 mg/kg神经降压素给药组：先使用LPS诱导肺损伤,诱导后1 h通过尾静脉注射的方式给予80 mg/kg神经降压素处理。每组20只。肺损伤诱导后24 h,检测不同处理组小鼠肺损伤程度、髓过氧化物酶（MPO）活性、炎症细胞的浸润、肺水肿程度以及肺泡灌洗液中促炎症细胞的分泌水平。 结果与正常对照组相比,LPS的滴鼻处理显著提高了小鼠肺组织的损伤程度,包括MPO活性、炎症细胞的浸润、肺泡灌洗液内促炎症细胞因子［肿瘤坏死因子（TNF-α）、白介素（IL）-6、IL-1b以及单核细胞趋化因子（MCP）-1］的分泌等（P<0.05）;与内毒素诱导的急性肺损伤模型组相比,神经降压素的给药处理明显减轻了小鼠肺组织损伤的程度,包括MPO活性、炎症细胞的浸润、肺泡灌洗液内促炎症细胞因子（TNF-α、IL-6、IL-1b以及MCP-1）的分泌等（P<0.05）,且这种保护性作用呈现剂量依赖性。 结论神经降压素能够有效减轻由内毒素诱导的ALI,其机制可能是通过封闭由速激肽所介导的炎症反应信号通路,进而减轻内毒素引起的炎症反应对肺组织所造成的炎症损伤来实现。     

白细胞介素基因多态性与急性胰腺炎病情程度的关系

目的 观察白细胞介素-1β(IL-1β)和IL-6基因多态性,探讨其与急性胰腺炎(AP)病情严重程度的关系.方法 采用聚合酶链反应一限制性片段长度多态性(PCR-RFLP)分析技术检测74例AP患者[轻型急性胰腺炎(MAP)36例,重症急性胰腺炎(SAP)38例]及78例健康对照者的IL-1β基因511C/T和IL-6基因634C/G位点多态性;采用酶联免疫吸附法(ELISA)检测血浆IL-1β和IL-6的浓度;SAP按不同基因型分组,比较各组间病情程度评分.结果 AP组血浆IL-1β、IL-6浓度高于健康对照组[IL-1β:(13.16±2.82)ng/L比(6.21±1.57)ng/L;IL-6:(84.86±32.92)ng/L比(9.95±2.49)ng/L,P均＜0.053.AP患者中,IL-1β基因511位点不同基因型血浆IL-1β浓度差异无统计学意义,IL-6基因634位点C/G基因型血浆IL-6浓度高于C/C型[(97.23±35.49)ng/L比(72.14±24.55)ng/L,P＜0.053.IL-1β基因511位点和IL-6基因634位点基因型及等位基因频率分布在AP组和健康对照组间差异无统计学意义;SAP组IL-1β基因511位点T/T型及T等位基因分布频率高于MAP组(P均＜0.05).SAP患者中,IL-1β基因511位点C/T+T/T型和IL-6基因634位点C/G型具有较高的病情程度评分.结论 IL-1β基因511C/T和IL-6基因634C/G位点多态性可能与AP的病情加重有关.     

白细胞介素-1β对小鼠气道成纤维细胞前列腺素E2表达的影响

目的研究白细胞介素-1β（IL-β）对小鼠气道成纤维细胞前列腺素E2（PGE2）表达的影响,以探讨其在吸入性损伤修复中的作用。方法分离雄性昆明种小鼠的气道成纤维细胞并进行体外培养,分别收集不同浓度的IL-1β作用后不同时间点IL-1β的培养上清液及细胞;采用酶联免疫吸附试验法和westem blot技术测定PGE2水平,环氧化酶-2（COX-2）、膜相关前列腺素E2合成酶1（mPGES1）蛋白表达。结果①经IL-1β1．0μg/L处理后不同时间点气道成纤维细胞培养上清液PGE2水平在8h[（148．2±28．3）ng／L]、16h[（267．6±45．4）μg／L]、24h[（210．5±38．6）ng／L]、48h[（198．7±36．5）ng／L]均高于各对照组[分别为（57．8±15．3）、（58．2±15．7）、（57．9±15．8）、（58．1±16．1）ng／L],差异均有统计学意义（P均〈0．01）,其中16h时间点水平最高;气道成纤维细胞COX-2表达在8h[（0．478±0．029）COX-2／β-actin、16h（0．672±0．047）cox-2／β-aetin、24h（0．617±0．036）cox-2／β-actin、48h（0．593±0．034）COX-2／β-actin]均高于对照组[（0．309±0．019）COX-2／β-actin、（0．311±0．019）COX-2／13-actin、（0．309±0．019）COX-2／β-actin、（0．310±0．018）cox-2／β-actin],差异有统计学意义（P均〈0．01）,其中16h时间点表达水平最高;气道成纤维细胞mPGESl的表达在8h（0．300±0．018）mPGES1／β-actin、16h（0．549±0．034）mPGES1／β-actin、24h（0．497±0．030）mPGES1/β-actin、48h（0．443±0．026）mPGES1／β-aetin均高于各对照组[（0．1994±0．012）、（0．201±0．013）、（0．200±0．013）和（0．200±0．012）mPGES1／β-actin],差异有统计学意义（P均〈0．01）,其中16h时间点表达水平最高。②不同浓度IL-1β处理气道成纤维细胞后,气道成纤维细胞培养上清液PGE：水平在IL-1β0．1μg／L组[（142．9±22．3）ng／L]、1．0μg／L组[（267．6±45．4）μg／L和10．0μg／L组[（587．3±106．9）μg／L]均高于对照组[（58．5±16．0）μg／L],差异有统计学意义（P均〈0．01）,并且这3组之间比较差异也有统计学意义（P均〈0．01）;气道成纤维细胞COX-2表达在IL-1β0．1μg／L组（0．525±0．031）ng／L、1．0μg/L组（0．672±0．047）ng／L和10.0μg/L组（1.012±0．064）ng／L均高于对照组（0．309±0．019）ng／L,差异有统计学意义（P均〈0．01）,并且这3组之间比较差异也有统计学意义（P均〈0．01）;气道成纤维细胞mPGES1表达在IL-1β0．1μg／L组（0．380±0．021）ng／L、1．0μg／L组（0．549±0．034）ng／L和10．0μg／L组（0．879±0．045）ng／L均高于对照组（0．199±0．012）ng／L,差异有统计学意义（P均〈0．01）,并且这3组之间比较差异也有统计学意义（P均〈0．01）。结论炎症递质IL-1β可上调气道成纤维细胞PGE2水平、COX-2和mPGES1表达,这表明IL-1β可能是通过COX-2和mPGES1的表达来影响PGE2合成,从而参与气道吸入性损伤修复过程。气道成纤维细胞可能是损伤修复过程中炎症递质的主要来源细胞之一。     

白细胞介素-1β对小鼠气道成纤维细胞前列腺素E_2表达的影响

目的 研究白细胞介素-1β(IL-1β)对小鼠气道成纤维细胞前列腺素E_2(PGE_2)表达的影响,以探讨其在吸入性损伤修复中的作用.方法 分离雄性昆明种小鼠的气道成纤维细胞并进行体外培养,分别收集不同浓度的IL-1β作用后不同时间点IL-1β的培养上清液及细胞;采用酶联免疫吸附试验法和West-ern blot技术测定PGE_2水平,环氧化酶-2(COX-2)、膜相关前列腺素E_2合成酶1(mPGES1)蛋白表达.结果 ①经IL-1β 1.0μg/L处理后不同时间点气道成纤维细胞培养上清液PGE_2水平在8 h[(148.2±28.3)ng/L]、16 h[(267.6±45.4)ng/L]、24 h[(210.5±38.6)ng/L]、48 h[(198.7±36.5)ng/L]均高于各对照组[分别为(57.8±15.3)、(58.2±15.7)、(57.9±15.8)、(58.1±16.1)ng/L],差异均有统计学意义(P均<0.01),其中16 h时间点水平最高;气道成纤维细胞COX-2表达在8 h[(0.478±0.029)COX-2/β-actin、16 h(0.672±0.047)COX-2/β-actin、24 h(0.617±0.036)COX-2/β-actin、48 h(0.593±0.034)COX-2/β-actin]均高于对照组[(0.309±0.019)COX-2/β-actin、(0.311±0.019)COX-2/β-actin、(0.309±0.019)COX-2/β-ac-tin、(0.310±0.018)COX-2/β-actin],差异有统计学意义(P均<0.01),其中16 h时间点表述水平最高;气道成纤维细胞mPGES1的表达在8 h(0.300±0.018)mPGES1/β-actin、16 h(0.549±0.034)mPGES1/β-actin、24h(0.497±0.030)mPGES1/β-actin、48 h(0.443±0.026)mPGES1/β-actin均高于各对照组[(0.199±0.012)、(0.201±0.013)、(0.200±0.013)和(0.200±0.022)mPGES1/β-actin],差异有统计学意义(P均<0.01),其中16 h时间点表达水平最高.②不同浓度IL-1β处理气道成纤维细胞后,气道成纤维细胞培养上清液PGE_2水平在IL-1β 0.1 μg/L组[(242.9±22.3)ng/L]、1.0μg/L组[(267.6±45.4)ng/L和10.0μg/L组[(587.3±106.9)ng/L]均高于对照组[(58.5±16.0)ng/L],差异有统计学意义(P均<0.01),并且这3组之间比较差异也有统计学意义(P均<0.01);气道成纤维细胞COX-2表达在IL-1β 0.1μg/L组(0.525±0.032)ng/L、1.0μg/L组(0.672±0.047)ng/L和10.0μg/L组(1.022±0.064)ng/L均高于对照组(0.309±0.019)ng/L,差异有统计学意义(P均<0.01),并且这3组之间比较差异也有统计学意义(P均<0.02);气道成纤维细胞mPGES1表达在IL-1β 0.1μg/L组(0.380±0.021)ng/L、1.0μg/L组(0.549±0.034)ng/L和10.0μg/L组(0.879±0.045)ng/L均高于对照组(0.199±0.022)ng/L,差异有统计学意义(P均<0.01),并且这3组之间比较差异也有统计学意义(P均<0.01).结论 炎症递质IL-1β可上调气道成纤维细胞PGE_2水平、COX-2和mPGES1表达,这表明IL-1β可能是通过COX-2和mPGES1的表达来影响PGE_2合成,从而参与气道吸入性损伤修复过程.气道成纤维细胞可能是损伤修复过程中炎症递质的主要来源细胞之一.     

高容量血液滤过对内毒素休克犬心肌组织Toll样受体4表达的影响

目的 探讨高容量血液滤过(HVHF)对内毒素休克心肌组织Toll样受体4(TLR4)mRNA表达的影响.方法 健康犬16只,静脉注射脂多糖(LPS)650 μg/kg诱导犬内毒素休克模型.将制摸成功的动物按随机数字表法分为对照组和HVHF治疗组,每组8只.用放射免疫法测定血中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-10含量,用逆转录-聚合酶链反应(RT-PCR)法测定心肌TLR4 mRNA表达,电镜下观察心肌组织病理改变.结果 治疗组HVHF后1、2、4 h血清TNF-α(μg/L:0.59±0.15、0.51±0.12、0.41±0.10)、IL-6(ng/L:11.08±2.83、9.82±2.58、8.25±2.05)、IL-10(μg/L:57.28±5.93、53.81±5.83、50.67±6.33)的含量显著低于本组成模时[(0.84±0.16)μg/L、(16.97±2.50)ng/L、(70.86±5.43)μg/L]和对照组相应时间点[TNF-α(μg/L):0.75±0.14、0.74±0.11、0.72±0.11,IL-6(ng/L):15.33±3.20、14.66±3.24、14.20±3.33,IL-10(μg/L):71.54±4.73、70.71±4.34、69.35±4.60],差异均有统计学意义(均P<0.01).治疗组较对照组心肌TLR4 mRNA表达明显下调(t=3.58,P<0.01).相关分析显示,TLR4 mRNA表达与循环血中TNF-α、IL-6、IL-10浓度呈正相关(r1=0.785,r2=0.569,r3=0.635,均P<0.05).电镜下观察治疗组心肌病理损伤较对照组减轻.结论 HVHF能下调内毒素诱导休克犬心肌TLR4mRNA 表达,减轻心肌炎症反应和心肌损伤.