羊栖菜褐藻糖胶CSFP-1抗肿瘤活性及机制研究

目的 分离纯化制备羊栖菜褐藻糖胶（CSFP-1）,并对其抗肿瘤活性及可能作用机制进行初步研究。方法 通过冷水浸提法制备羊栖菜褐藻糖胶（CSFP）,经超滤系统分离获得CSFP-1,并采用苯酚-硫酸比色法、PMP衍生化、高效液相色谱法、红外光谱分析等多种方法对其理化性质进行分析;采用MTT法评价CSFP-1（0、10、100、300 μg/mL）对宫颈癌细胞（HeLa）、肺癌细胞（A549）和肝癌细胞（Hep3B）生存能力的影响;应用流式细胞术评价CSFP-1对HeLa细胞细胞周期的影响;应用Western blotting技术评价CSFP-1对HeLa细胞凋亡和自噬相关蛋白表达的影响。结果 CSFP-1的糖含量为84.18%,糖醛酸含量为14.28%,硫酸根含量为10.02%,平均相对分子质量为5.6×10⁴,主要由半乳糖、岩藻糖、甘露糖、葡萄糖醛酸和木糖组成。CSFP-1具有选择性细胞毒性,对HeLa细胞生长发挥显著抑制作用（P<0.05、0.001）,对A549和Hep3B细胞存活率没有明显影响。与对照组比较,HeLa细胞经CSFP-1处理24 h后,G1期细胞比例明显增加（P<0.001）;经CSFP-1处理48 h后,S期细胞比例显著增加（P<0.05、0.01、0.001）,G2期细胞比例显著减少（P<0.01、0.001）,均呈现剂量相关性。与对照组比较,CSFP-1 100、300 μg/mL剂量组处理24 h,300 μg/mL剂量组处理48 h Bcl-2蛋白表达显著减少（P<0.05）;100、300 μg/mL剂量组处理24、48 h Beclin-1蛋白表达显著增加（P<0.05、0.01）,且存在时间和剂量相关性。结论 CSFP-1具有选择性抑制HeLa细胞增殖作用,作用机制可能与阻滞细胞周期、诱导细胞凋亡相关。     

Analysis of Adhesion Molecules Involved in Leukocyte Homing into the Basolateral Pockets of Mouse Peyer's Patch M Cells

The basolateral membranes of intestinal M cells are invaginated to form large intraepithelial “pockets” that are populated by specific sub-sets of mucosal leukocytes, including CD4+ T cells, memory and naïve B cells, and occasional dendritic cells. The adhesion molecules involved in leukocyte trafficking and/or retention within this unique immunological niche are unknown. In this study, we used immunofluorescence microscopy and a battery of monoclonal antibodies to identify the adhesion molecules expressed by leukocytes situated within the intracellular pockets of mouse Peyer's patch (PP) M cells. M cell associated leukocytes (MAL) consistently stained positive for integrin α4β7, and integrin LFA-1 (CD11a/CD18), but were rarely positive for L-selectin (CD62L) or the mucosal integrin αEβ7. However, neither the α4β7 ligands MadCAM-1 or VCAM-1, nor the LFA-1 ligand ICAM-1, were detected on M cell basolateral membranes. To determine whether integrins α4β7 or LFA-1 play a functional role leukocyte homing to M cell pockets, we examined M cells in mice deficient in integrin β7 or CD11a/CD18. Although PP from CD18_-/- or integrin β7_-/- mice were reduced in number and size as compared to age-matched controls, we identified M cells in both strains of mice. However, mice lacking CD18 (but not integrin β7) had significantly fewer leukocytes within M cell pockets as compared to control animals, suggesting LFA-1 (but not α4β7) may contribute, in part, to leukocyte trafficking into and/or retention within this unique immunological niche.     

大鼠过敏性气道炎症中VCAM—1表达、嗜酸细胞浸润和药理调节

目的:探讨过敏性气道炎症大鼠肺内VCAM-1表达和嗜酸细胞浸润,以及速激肽NK-1受体拮抗剂SR140333和地塞米松的可能调节方式.方法:致敏大鼠以1％卵白蛋白气雾吸入攻击后24小时,测定VCAM-1在不同组别大鼠肺内的表达和支气管周围嗜酸细胞浸润.抗原攻击前3天,每天2次腹腔注射SR140333(0.01,0.10mg/kg)或地塞米松(0.50mg/kg).结果:与未致敏大鼠相比VCAM-1在致敏大鼠肺内表达增加(P<0.05),预先用地塞米松处理可抑制其增加(P<0.05),SR140333无此作用(P>0.05).此外,抗原攻击致敏大鼠促使支气管周围嗜酸细胞浸润,但不能进一步增加VCAM-1的表达(P>0.05).SR140333抑制嗜酸细胞浸润(P<0.05),但不影响VCAM-1表达(P>0.05);地塞米松则抑制这两种反应(P<0.05).结论:VCAM-1表达在抗原致敏后增加,地塞米松和SR140333抑制大鼠过敏性气道炎症的机制可能不同.     

Bothrops jararaca venom (BjV) induces differential leukocyte accumulation in mice genetically selected for acute inflammatory reaction: The role of host genetic background on expression of adhesion molecules and release of endogenous mediators

The dynamics of the local inflammatory events induced by Bothrops jararaca venom (BjV) inoculation in footpad of mice genetically selected for maximal (AIRmax) and minimal (AIRmin) acute inflammatory reactivity (AIR) was investigated. The BjV injection induced a marked inflammatory cell infiltrate with predominance of neutrophils, with increased blood cell numbers before its accumulation, suggesting a stimulatory action of BjV on mechanisms of cell mobilization from bone marrow. The process of cell migration is regulated by different cell-adhesion molecules (CAM). Our results showed that neutrophil cells from both lines had the same pattern of response concerning CAMs expression, presenting the involvement of l-selectin, Mac-1 and PECAM-1 adhesion molecules in BjV-induced neutrophil accumulation. The effect of BjV on the release of pro-inflammatory cytokines and chemokines related with cellular migration was also studied and IL-1β, IL-6, TNF-α and MIP-2 levels could be detected after venom injection. The AIRmax mice were shown to be more responsive than AIRmin with respect to leukocyte influx, expression of MIP-2 and release of IL-1β and IL-6. These results demonstrate the importance of host genetic background in the local response and the involvement of alleles accumulated in AIRmax mice in the inflammatory events induced by BjV.     

茶碱对哮喘气道炎症的作用(英文)

目的:研究小剂量茶碱对哮喘气道炎症的作用。方法:19名哮喘患者用茶碱缓释剂(200mg,bid,平均血浆茶碱浓度7.9mg/L)治疗4周,分别用瑞氏染色、免疫组织化学及荧光免疫法检测治疗前后高渗盐水诱导痰中嗜酸细胞(Eos)、激活的Eos(EG2~(2 )Eos)和嗜酸细胞阳离子蛋白(ECP)的变化,并观察治疗前后症状积分和肺功能的变化。结果:用茶碱治疗前,患者痰中Eos、EG~(2 )Eos和ECP比健康人明显增加;用茶碱治疗4周后,哮喘患者诱导痰中Eos百分数下降(40％±17％ vs 29％±11％,P<0.01),EG~(2 )Eos百分数显著下降(28％±9％ vs 10 ％±8％,P<0.01),痰ECP明显下降[(373±206)vs(220±132)μg/L,P<0.01];症状明显好转(7.1±1.2 vs 5.4±1.6,P<0.01);肺功能明显改善,FEV_(1.0)增加(2.2±0.6 vs 2.4±0.5,P<0.01),FEV_(1.0)％也增加(60％±13％ vs 65％±13％,P<0.01)。结论:小剂量茶碱对哮喘气道炎症有明显抑制作用,同时使患者症状和肺功能明显改善。     

黏附分子在EMT及癌细胞干性调节中的分子机制研究进展

细胞黏附能力及能动性的变化在转移性肿瘤发生发展过程中起着关键作用。上皮-间质转化主要发生在胚胎发育期和癌细胞间黏附特性变化的过程中,并且该过程被认为是转移性肿瘤进展的主要途径。近期有关循环肿瘤微栓子的研究指出,在肿瘤进展期间上皮-间质转化（EMT）是逐步的而不是“全有或无”的二元转变。本文综述了细胞黏附分子及其分子机制调控不完全上皮-间质转化状态及癌细胞干性的分子机制。     

Effect of in vivo phenol or hydroquinone exposure on events related to neutrophil delivery during an inflammatory response

Phenol (PHE) and hydroquinone (HQ) are metabolites of benzene that affect leukocytes after solvent intoxication. Hence, we investigated the effects of PHE or HQ exposure on neutrophil mobilization during an inflammatory response. Male Wistar rats received intraperitoneal injections of PHE, HQ or vehicle only and assays were performed 24 h after the last dose. Quantifications of bone marrow or circulating leukocytes showed that only HQ exposure induced neutrophilia, probably due to the accelerated mobilization from the bone marrow compartment, since reduced numbers of segmented cells in the last phase of maturation were detected there. Intravital microscopy showed that circulating leukocytes of HQ-exposed rats increased their rolling behavior and adherence to the mesenteric postcapillary venule wall in vivo. The enhanced leukocyte–endothelium interaction was not dependent on microvascular reactivity or perivascular mast cell degranulation. Instead, it was the result of neutrophil activation, demonstrated by a decrease in l-selectin and an increase in β₂ integrin expression on neutrophil membranes. This pattern of neutrophil activation may have contributed to the higher number of neutrophils in the subcutaneous inflammatory response of HQ-exposed rats after oyster glycogen injection. Taken together, our results indicate that HQ exposure alters neutrophil mobilization, which results in an exacerbated response after an injury. Although PHE is endogenously metabolized to HQ, PHE exposure only induced an increment in rolling behavior, which was not sufficient to alter the inflammatory response.     

A novel murine model of allergic inflammation to study the effect of dexamethasone on eosinophil recruitment

1. We have developed a novel model of allergen-induced eosinophil extravasation into mouse air-pouches following sensitization and challenge with ovalbumin (Ova). This model was used to investigate the mechanism(s) underlying the anti-inflammatory action of the glucocorticoid hormone dexamethasone (Dex).
2. Injection of 10 μg Ova into 6-day-old dorsal air-pouches of mice sensitized to the same antigen provoked an intense cell accumulation as early as 6 h post-challenge (0.08±0.03 and 4.0±1.0×10⁵ leucocytes in saline and Ova-treated air-pouches, respectively), maximal at 24 h (0.02±0.01 and 6.0±0.8×10⁵ leucocytes in saline and Ova-treated air-pouches, respectively) and persisted up to 48 h. At the 24 h time-point, the cellular infiltrate consisted of 37% eosinophils, 18% neutrophils and 45% mononuclear cells, as assessed by histological examination. The same ratio of eosinophil/neutrophil was obtained by fluorescence-activated cell sorting (FACS) analysis, since 72% of the polymorphonuclear (PMN) population was positive for very-late antigen-4 (VLA-4) expression.
3. Subcutaneous (s.c.) administration of Dex (50 or 100 μg per mouse, −1 h) inhibited eosinophil accumulation into Ova challenged air-pouches by about 70% (P<0.05) and 75% (P<0.05), respectively, when compared to controls. Cell accumulation measured at 48 h after Ova injection was also significantly reduced (−75%) by Dex administration at the 24 h time-point (n=12, P<0.05).
4. Eosinophil numbers in the bone marrow and blood were quantitated. We found that the sensitization protocol induced a 3 fold increase in eosinophil numbers in the bone marrow (naive mice: 2.7±0.3×10⁵; sensitized mice: 8.7±1.7×10⁵, P<0.05) and blood (naive mice: 0.5±0.2×10⁵; sensitized mice: 1.5±0.3×10⁵, P<0.01). However, 24 h following Ova challenge, the eosinophil numbers in the bone marrow had dropped (3.7±0.8×10⁵) with no change in the circulating pool, suggesting an equilibrium within the eosinophil pools had been reached.
5. Dex administration provoked a profound eosinopaenia in the blood of naive (5.2±1.5 to 0.9±0.6×10⁴) and sensitized mice (1.5±0.3 to 0.08±0.02×10⁵) at 4 h. This effect was reversed within 24 h. Dex also inhibited the release of eosinophils from the bone marrow in response to Ova challenge.
6. We show for the first time that eosinophils express the steroid-inducible protein lipocortin 1 (LC1). FACS analysis of eosinophils emigrated into the Ova challenged air-pouches revealed detectable LC1-like immunoreactivity (373×10³). These data were also substantiated by LC1 detection in circulating eosinophils of interleukin-5 transgenic mice (strain: CBA/Ca). However, s.c. injection of Dex (50 μg) did not alter LC1 levels in blood eosinophils, such that 235±21×10³ LC1-like molecules per cell were measured after vehicle treatment (n=5), and 224±8×10³ LC1-like molecules per cell were associated with this cell type 1 h after steroid treatment (n=5, not significant). Finally, resident eosinophils (in the pleural cavity) were found to have much higher LC1 levels than that found in the blood circulation (2 fold increase, P<0.05).
7. Passive immunization of mice against LC1 with a validated antiserum (termed LCS3) and protocol failed to modify the anti-migratory activity exerted by Dex towards eosinophil extravasation into Ova-challenged air-pouches. The steroid (50 μg s.c., −1 h) produced a similar degree of inhibition of eosinophil accumulation both in control animals (treated with a non-immune sheep serum) and LCS3-treated mice (−56% and 59%, respectively, n=15–21, not significant).
8. In conclusion, the air-pouch provides a novel and convenient cavity to study allergen-induced cell recruitment which is sensitive to glucocorticoid hormone treatment. The effect of Dex on eosinophil distribution in these experimental conditions has been studied in detail and we failed to find an important role for endogenous LC1 in these actions of the steroid.

     

Antioxidant Activity of Plicatin B on Cultured Human Microvascular Endothelial Cells Exposed to H2O2

Oxidative stress plays a crucial role in the pathogenesis of atherosclerosis by promoting endothelial dysfunction and impairing vascular relaxation. Flavonoids are largely investigated for their biological properties and particularly for their scavenging and antioxidant properties. In the current study, we evaluated the clastogenicity of the chalcone plicatin B in peripheral human lymphocytes (whole blood and pure lymphocytes) as well as its antioxidant activity and its ability to contrast dysfunction on human microvascular endothelial cells (HMEC-1) exposed to hydrogen peroxide. We measured in the cell culture medium the levels of 8-isoprostane, NO_x, ET-1, and ICAM-1, as well as the expression of e-NOS, prepro-ET-1, and ICAM-1. In conclusion, our results demonstrate that the chalcone plicatin B (1–10 μM) may represent a good candidate for the prevention of atherosclerosis, as it consistently reduces the oxidative/inflammatory process and is not genotoxic to human lymphocytes.     

丹参对实验糖尿病大鼠粘附分子CD54 CD106 CD62p的影响

目的　研究丹参对链脲佐菌素 (STZ)诱导的糖尿病大鼠模型体内三种粘附分子 (CD5 4、CD10 6、CD6 2 p)的变化和对肾脏、主动脉病理变化的影响。方法　随机分正常对照、糖尿病、胰岛素、丹参四组 ,测定各组血糖、糖化血红蛋白、单个核细胞表面CD5 4、血小板表面CD6 2 p表达水平 ,观察肾脏和大血管CD5 4和 /或CD10 6表达强度及病理变化。结果　丹参降低肾脏CD5 4、CD10 6 ,主动脉CD10 6的表达 ,也降低单个核细胞CD5 4的表达(丹参组 30± 2 5vs糖尿病组 6 5± 15 ,P <0 0 5 ) ,但对血小板CD6 2 p的表达无明显影响 ,肾脏及主动脉的病理变化明显减轻。结论　丹参能降低糖尿病大鼠体内CD5 4、CD10 6水平 ,并能改善糖尿病引起的肾脏和主动脉的病理变化     

Differential surface expression of CD18 and CD44 by neutrophils in bone marrow and spleen contributed to the neutrophilia in thalidomide-treated female B6C3F1 mice

Previously, we have reported that thalidomide (Thd) can enhance neutrophil function in female B6C3F1 mice. The present study was intended to evaluate the mechanisms underlying the enhanced neutrophil responses following Thd treatment intraperitoneally (100 mg/kg) for 14 or 28 days. Treatment with Thd increased the numbers of neutrophils in the spleen, peripheral blood, bone marrow, peritoneal cavity and lungs of female B6C3F1 mice when compared to the vehicle control mice. Thd treatment for 14 days increased the percentage and the number of neutrophils in the spleen in the first 8 h (peaking at 2 h) after the last Thd treatment, and it returned to the baseline after 24 h. However, Thd treatment for 28 days increased the percentage and number of neutrophils in the spleen even at the 24-h time point after the last Thd treatment. These neutrophils were demonstrated to be functional by the myeloperoxidase activity assay. Further studies have ruled out the possibility of an increased bone marrow granulopoiesis following Thd treatment. Flow cytometric analysis of the surface expression of adhesion molecules suggested that Thd treatment for either 14 or 28 days decreased the surface expression of either CD18 or CD44 by bone marrow neutrophils. On the other hand, the surface expression of both CD18 and CD44 by splenic neutrophils was increased following Thd treatment for 28 days but not for 14 days. No effect was produced for other cell surface molecules such as CD62L and CD11a. It was possible that decreased surface expressions of CD18 and CD44 facilitated neutrophils' release from the bone marrow; increased surface expressions of CD44 and CD18 by splenic neutrophils after 28 days of Thd treatment increased their ability to remain in the periphery. Taken together, Thd treatment increased neutrophils in female B6C3F1 mice, at least partially, through differentially modulating the surface expression of CD18 and CD44 by the neutrophils in the bone marrow and spleen.     

Protective effect of phosphorylated Athyrium multidentatum (Doll.) Ching polysaccharide on vascular endothelial cells in vitro and in vivo

The purpose of this study was to prepare phosphorylated Athyrium multidentatum (Doll.) Ching polysaccharide (PPS) and investigate its protective effect on vascular endothelial cells (VECs) in vitro and in vivo and the underlying mechanisms. Sodium tripolyphosphate (STPP) and sodium trimetaphosphate (STMP) were used as phosphorylation reagents and PPS was characterized by Fourier transform infrared (FT-IR), ¹³C nuclear magnetic resonance (¹³C NMR) and ³¹P nuclear magnetic resonance (³¹P NMR) spectra. Chemical analysis demonstrated that PPS was composed of mannose, glucosamine, rhamnose, glucuronic acid, galacturonic acid, galactosamine, glucose, galactose, xylose, arabinose, and fucose with a molar ratio of 11.36:0.42:4.03:1.12:1.81:0.26:33.25:24.12:6.85:14.46:2.32 and a molecular weight of 28,837 Da. Results from in vitro and in vivo assays revealed that PPS protected human umbilical vein endothelial cells (HUVECs) against H₂O₂-induced oxidative injury and attenuated D-galactose-induced VECs damage in mice. RNA sequencing (RNA-seq) analysis identified 18 differentially expressed genes (DEGs) between D-galactose-treated and PPS-pretreated mice abdominal aorta. A deep analysis of these DEGs disclosed that PPS regulated the expression of genes involved in the functions of vascular endothelium repairment, cell growth and proliferation, cell survival and apoptosis, inflammation, angiogenesis and antioxidant, indicating that these biological processes might play crucial roles in the protective actions of PPS on VECs.     

白细胞介素-1受体拮抗剂对卵白蛋白致敏的哮喘豚鼠嗜酸性粒细胞凋亡的影响

目的研究白细胞介素-1受体拮抗剂(IL-1ra)，对慢性哮喘豚鼠肺内嗜酸性粒细胞凋亡及相关机制的影响。方法建立致敏豚鼠模型，连续吸入不同浓度IL-1ra,用卵白蛋白引喘豚鼠，连续8 d，分离血液、肺泡灌洗液(BALF)中嗜酸性粒细胞，计数嗜酸性粒细胞，用放射免疫法测定上清液中阳离子蛋白（ECP），酶连免疫法（ELISA）测血清中IL-5，荧光显微镜观察嗜酸性细胞凋亡。结果IL-1ra能明显减少肺部嗜酸性粒细胞浸润，降低血清、肺泡灌洗液中ECP水平及血清中IL-5的含量，促进嗜酸性细胞凋亡。结论雾化吸入IL-1ra可以通过改变嗜酸性粒细胞的活性及浸润程度，达到治疗哮喘的目的。     

肺成纤维细胞对嗜酸性粒细胞存活率的影响

目的 研究肺成纤维细胞(FB)对健康人外周血嗜酸性粒细胞(EOS)存活率的影响,并探讨其作用机制。方法 将FB与EOS共同培养,采用台盼蓝拒染法检测EOS存活率、酶联免疫吸附法(ELISA)检测上清液中粒细胞-巨噬细胞集落刺激因子(GM-CSF)含量。结果 (1)与EOS单独培养相比,FB和EOS共同培养后EOS存活率明显上升,两组相比有显著性差异(P<0.001),经0.2％戊二醛固定后的FB对EOS存活率无明显影响。(2)FB和EOS共同培养后GM-CSF含量明显增加,与其他各组相比均有显著性差异。结论 (1)肺FB在体外能明显提高外周血EOS存活率。(2)FB和EOS的共同培养可以导致细胞因子GM-CSF增加,这可能是FB延长EOS存活率的作用因素之一。     

Sympathoadrenal-dependent sexually dimorphic effect of nonhabituating stress on in vivo neutrophil recruitment in the rat

Since stress both activates the sympathoadrenal axis and profoundly affects inflammation and inflammatory diseases, many of which are sexually dimorphic, we tested whether the effect of stress on neutrophil recruitment, a primary component of the acute inflammatory response, is sexually dimorphic. The effect of intermittent sound (over 4 days), a nonhabituating stress, on lipopolysaccharide (LPS)-induced recruitment of neutrophils was evaluated in vivo in the rat air pouch model. At 24 h following the last stress exposure, LPS-induced neutrophil recruitment was enhanced in male rats, but not in females. When gonadectomized prepubertally and tested as adults, stress significantly inhibited the magnitude of LPS-induced neutrophil recruitment in males, while it still had no effect in gonadectomized females. In males, following adrenal denervation, the increase in LPS-induced neutrophil recruitment produced by stress was prevented. Since these data suggest that the effect of stress is dependent on the sympathoadrenal axis, we tested the hypothesis that catecholamines mediate the stress effects.In male rats, the effect of stress on LPS-induced neutrophil recruitment was significantly attenuated by continuous administration of the beta-adrenergic receptor antagonist, propranolol (4 mg kg(-1) day(-1)), during sound stress exposure, and administration of isoproterenol (10 nmoles, i.v.) significantly increased neutrophil recruitment in males, an effect that was qualitatively and quantitatively similar to the effect of stress. Propranolol significantly increased neutrophil recruitment in nonstressed female rats, but did not significantly affect neutrophil recruitment in stressed females. These findings indicate a marked male sex hormone-dependent sexual dimorphism in the sympathoadrenal-dependent effect of stress on neutrophil migration, a primary component of the inflammatory response, and suggest that the sympathoadrenal axis contributes to this effect via release of epinephrine.     

Research on pharmacological mechanism of the treatment of Asthma by oral Bordetella pertussis

Objective To examine the effect of oral Bordetella pertussis on the asthma mice sensitized by ovalbumin(OVA),and explore the possible mechanism.Methods Culture the B.pertussis in Bordet-Gengou agar containing 25% rabbit blood.Collect the bacteria and inactive them at 80 ℃ for 30 min to get whole killed B.pertussis.32 BALB/C mice were randomly divided into control group,model-control group,model group and treatment group.The mice were sensitized and challenged with OVA to establish asthma model.Asthma mice in treatment group were orally administrated with B.pertussis 7 days before sensitization.The mice in control group and model-control group were challenged with saline.After 24 hours of last challenge,bronchoaveolar lavage fluid(BALF)and peripheral blood were collected.The total cells and eosinophils were counted in BALF.Results Compared with the control group(2.03±0.42,0.33±0.82)×105 mL-1 and model-control group(2.16±0.48,0.16±0.41)×105 mL-1,the total cells(10.13±1.33)×105 mL-1 and eosinophils(11.83±4.573)×105 mL-1 in BALF were more in asthma mice(P<0.01).The number of total cells(5.50±1.55)×105 mL-1 and eosinophils(0.66±0.82)×105 mL-1 in BALF were reduced in asthma mice treated with B.pertussis compared with asthma mice(P<0.01).Conclusions Oral B.pertussis can inhabit airway inflammation of asthma mice and has the potential of treating asthma.     

s-油酰丙醇胺对人脐静脉内皮细胞黏附分子表达的影响

目的探讨s-油酰丙醇胺对用肿瘤坏死因子α(TNF-α)诱导的人脐静脉内皮细胞黏附分子(VCAM-1,I-CAM-1,E选择素)表达的影响。方法从新鲜的脐带中分离出人脐静脉内皮细胞,培养至3~9代,用不同浓度的s-油酰丙醇胺(10,50,100μmol/L)孵育12 h后,用TNF-α(20 ng/mL)孵育8 h,采用荧光实时定量PCR和细胞酶联免疫吸附试验分别检测VCAM-1,ICAM-1,E选择素的mRNA及蛋白的表达,同时采用细胞黏附实验检测其对细胞黏附的影响。结果相对于正常的人脐静脉内皮细胞,TNF-α诱导后的人脐静脉内皮细胞黏附分子(VCAM-1,ICAM-1,E-选择素)的表达明显增加。s-油酰丙醇胺可以显著的抑制VCAM-1的表达,并呈现出一定的剂量依赖性,而且对人急性单核细胞性白血病细胞(THP-1)的黏附也有明显的抑制作用,但对ICAM-1,E-选择素的表达却没有影响。结论s-油酰丙醇胺和大多数的PPARα激动剂一样,能够抑制慢性炎症,减少单核细胞的黏附,抑制VCAM-1的表达,而对急性炎症没有作用,如对E-选择素的表达无影响。     

己酮可可碱在大鼠肠缺血再灌注损伤中的保护作用及机制

目的:探讨己酮可可碱(pentoxifylline,PTX)在大鼠肠缺血再灌注损伤中的保护作用及机制。方法:Wistar大鼠48只随机分成6组,每组8只。(1)假手术组(S); (2)缺血组(I); (3 )缺血再灌注2h组(IR2h); (4)缺血再灌注4h组(IR4h);(5)缺血再灌注2h+PTX组(P2 ); (6)缺血再灌注4h+PTX组(P4 ),测定各组动物血清肿瘤坏死因子α(TNF α)水平、肠组织细胞粘附分子(ICAM 1 )表达、肠组织过氧化物酶(MPO)活性及观察病理形态。结果:IR2h及IR4h组大鼠血清TNF α水平、肠组织ICAM 1表达及肠MPO活性均明显高于S组(P<0. 01)。给予PTX后, 血清TNF α水平有所下降,P4 组与IR4h组比较差异有非常显著意义(P<0. 01);肠组织MPO活性明显降低,IR2h组与P2 组比较或IR4h组与P4 组比较差异均有非常显著意义(P<0. 01);肠组织ICAM 1积分光密度值比较,P2,P4 组明显低于IR2h及IR4h组(P<0. 01)。结论:已酮可可碱可降低血TNF α水平及肠组织ICAM 1表达,减轻中性粒细胞在肠内聚集、活化,从而防治肠缺血再灌注损伤。