甲泼尼龙与抑肽酶对心肺转流术致全身炎症反应过程中白细胞磷脂酶D活性的影响

目的:探讨白细胞磷脂酶D(PLD)在心肺转流术(CPB)致全身性炎症反应过程中的作用及甲泼尼龙和抑肽酶对PLD活性的影响。方法:42例接受CPB心脏直视手术病人,分为对照组、甲泼尼龙组和抑肽酶组,患者于术前、心肺转流术中及术后8个不同时点采集动脉血,测定白细胞PLD活性,并检测髓过氧化物酶活性、CD11b表达及血浆IL-6、IL-8和C-反应蛋白的含量。结果:在升主动脉开放时点,对照组白细胞PLD活性为(18±8)nmol choline·h~(-1)·mg~(-1),明显高于术前;甲泼尼龙组为(10±6)nmol choline·h~(-1)·mg~(-1),明显低于对照组,但与术前相比差别无显著意义;甲泼尼龙组白细胞PLD活性升高时点后移至停CPB即刻点;抑肽酶组PLD活性与对照组相比差别无显著意义;但两药均不同程度降低CPB围术期血浆中IL-6、IL-8和C-反应蛋白水平及MRO活性、白细胞CD11b表达。结论:CPB致全身炎症反应过程中白细胞的PLD活性持续升高;甲泼尼龙部分抑制CPB引起的炎症反应,其抗炎机制与抑制PLD有关。     

荆芥挥发油对胸膜炎模型大鼠的抗炎作用研究

目的:研究荆芥挥发油对急性胸膜炎模型大鼠的抗炎作用及机制,为其祛风散寒之功效与治疗表证的临床应用提供药理学依据.方法:采用胸腔注射角叉菜胶制备大鼠急性胸膜炎模型,荆芥挥发油(0.4、0.2 mL·kg-1)于致炎前后1小时分别灌胃给药1次,造模后5小时处死大鼠,测定胸腔渗出液体积,进行渗出液中白细胞计数与分类、蛋白质含量测定来观察药物的抗炎作用;测定渗出液中前列腺素E2(PGE2)、白细胞介素1(IL-1)与肿瘤坏死因子(TNF-d)含量以及肺组织匀浆中丙二醛(MDA)、髓过氧化物酶(MPO)含量,来观察药物的抗炎作用机制.结果:荆芥挥发油对模型大鼠胸腔炎性渗出液体积有降低趋势,能显著降低渗出液中白细胞计数与蛋白质含量;能显著降低渗出液中炎症介质PGE2、IL-1与TNF-α含量,同时对模型大鼠肺组织中MDA的生成有明显抑制作用,并表现出降低MPO含量的趋势.结论:荆芥挥发油对角叉菜胶所致大鼠急性胸膜炎具有良好的抗炎作用,其作用机制与抑制炎症介质的生成及抗氧化作用有关.     

磷脂酶D与胞吞、胞吐的关系探讨

磷脂酶D(phospholipase D，PLD)可分为PLD1和PLD2两大类，最早发现于植物，后相继于细菌、真菌以及哺乳动物中被发现。该酶的活性可为各种激素、生长因子以及其它细胞外信号所调节。PLD可酶解其最主要的底物磷脂酰胆碱(phosphatidylcho1ine，PC)，生成磷脂酸(phosphatidic acid，PA)和胆碱，而前者又可在磷脂酸水解酶(phosphatidic acid phosphatase，PAP)以及特异性的磷脂酶A2(phospholipase A2，PLA2)的作用下生成甘油二酯(diacylglycerol，DG)和溶血磷脂酸(1ysophosphatidic acid，LPA)。PLD还可催化转磷脂酰基反应生成磷脂酰醇(phosphatidly alcohol)，此乃一稳定的产物，其性质被用于对PLD的活性等进行分析。     

革兰阴性菌磷脂酶D致病机制的研究进展

磷脂酶D (PLD)是致病因子的一种,通过多种机制帮助病原菌感染和复制,其有望成为治疗病原菌感染的新靶标.本文综述几种革兰阴性菌PLD致病机制的研究进展.     

水杨酸甲酯糖苷抗大鼠急性胸膜炎的作用研究

目的水杨酸甲酯糖苷(DL0309)是来源于民族药滇白珠的新型非甾体抗炎药,本实验的主要目的是评价其对角叉菜胶致大鼠胸膜炎模型的抗炎作用及其可能的机制。方法将48只♂SD大鼠按体重随机分为正常对照组、模型对照组、阳性对照组(地塞米松)、DL0309低、中、高剂量组。通过注射角叉菜胶,建立大鼠急性胸膜炎模型。造模后5 h处死大鼠,通过测定胸腔渗出液的体积,对渗出液中的白细胞计数,并测定渗出液中蛋白质的含量来观察药物对该模型的抗炎作用;通过测定渗出液中一氧化氮(NO)、肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)和前列腺素E2(PGE2)以及血浆中超氧化物歧化酶(SOD)和丙二醛(MDA)的含量,来考察DL0309对该模型的抗炎作用机制。结果结果显示地塞米松和DL0309均可明显降低胸膜炎大鼠胸腔炎性渗出液的体积以及渗出液中的白细胞数量和蛋白质含量,同时对于胸腔炎性渗出液中的NO、TNF-α、IL-1β和PGE2的含量均有不同程度的抑制,能不同程度地降低血浆中MDA含量,升高SOD活力。结论 DL0309具有抗角叉菜胶诱导的大鼠急性胸膜炎作用。     

鼻咽癌患者糖基化磷脂酰肌醇特异性磷脂酶D 活性及其表达水平的初步研究

目的①通过研究正常人和鼻咽癌患者的糖基化磷脂酰肌醇特异性磷脂酶D（GPI-PLD）的酶活性和酶促反应的3-D图,探索鼻咽癌患者该酶活性的变化情况及该酶促反应的可能机制。②通过研究正常人和鼻咽癌患者GPI-PLD mRNA的表达水平,探寻鼻咽癌患者该酶活性变化的可能机制,为鼻咽癌的诊断、预后估计提供有效指标。方法①采用我室自制的具完整GPI结构的胎盘型碱性磷酸酶（PLAP）作底物进行酶促反应,测定正常人和鼻咽癌患者GPI-PLD的酶活性并应用韩国新科有限公司生产的Photodiode Array Uv-Vis Spectrophotometer仪器研究了该酶的催化机理,并且利用该机所带软件作出3-D图。②采用逆转录PCR（RT-PCR）检测正常人和鼻咽癌患者的GPI-PLD mRNA的表达水平,以RT-PCR结果的琼脂糖电泳照片的积分光密度值（IA）比值代表GPI-PLD mRNA的表达水平。结果①正常人和鼻咽癌患者血清GPI-PLD的活性水平（均以转化底物百分比表示）分别为17.6%±2.7%和20.6%±2.4%,鼻咽癌患者的血清GPI-PLD活性比正常人血清GPI-PLD活性显著增高（P〈0.05）。并且其酶促反应的3-D图显示有直观差异。2.RT-PCR检测GPI-PLD mRNA的表达,结果表明正常人鼻咽上皮细胞中GPI-PLD mRNA琼脂糖电泳照片的平均积分光密度比值（IA比值）是2.6131±0.7653,鼻咽癌患者鼻咽上皮细胞中GPI-PLD mRNA琼脂糖电泳照片的平均积分光密度比值（IA比值）是9.3522±5.2879,鼻咽癌患者的GPI-PLD mRNA的表达水平比正常人鼻咽上皮细胞中GPI-PLD mRNA的表达水平显著性增高（P〈0.05）。结论①鼻咽癌患者的血清GPI-PLD活性比正常人显著增高,其酶促反应的3-D图也与正常人有明显差异,提示测定GPI-PLD活性可作为临床诊断鼻咽癌的生化指标。②鼻咽癌患者GPI-PLD mRNA的表达水平高于正常人。GPI-PLD mRNA表达增强是导致鼻咽癌患者GPI-PLD酶活性增高的主要机制。     

沙丁胺醇与色甘酸钠对大鼠腹腔肥大细胞脱颗粒过程中磷脂酶D活性的影响

采用主动致敏的大鼠腹腔肥大细胞 (RPMC)为实验材料 ,细胞脱颗粒以 β 氨基己糖苷酶释放率为指标 ,细胞磷脂酶D(PLD)活性采用酶联化学发光法检测 .结果发现 ,卵白蛋白 (4mg·L- 1)攻击主动致敏的RPMC 12 0s后 ,细胞PLD活性与β 氨基己糖苷酶释放率均明显增加 ,分别为对照组的 2倍及 8倍以上 .1%正丁醇 ,10mmol·L- 12 ,3 二磷酸甘油酸能显著抑制PLD活性 ,且使PLD活性及细胞 β 氨基己糖苷酶释放率均降低到对照水平 .Wortmannin ,30 0nmol·L- 1能显著抑制PLD活性 ,并减少细胞 β 氨基己糖苷酶释放 .1,10 μmol·L- 1的沙丁胺醇与色甘酸钠均能部分抑制PLD活性 ,并减少细胞 β 氨基己糖苷酶的释放 .结果表明 ,PLD在主动致敏的RPMC脱颗粒过程中起一定作用 ;沙丁胺醇 (1,10 μmol·L- 1) ,色甘酸钠 (1,10 μmol·L- 1)抑制主动致敏的RPMC脱颗粒的作用机理与抑制PLD有关 .     

玉郎伞提取物对角叉菜胶致大鼠急性胸膜炎的影响

目的观察玉郎伞(YLS)提取物对角叉菜胶诱导的大鼠胸膜炎的影响。方法56只Wistar雄性大鼠随机分为正常组,模型组,地塞米松组,YLS水提取物(TYLS)高、低剂量组,总黄酮(FYLS)高、低剂量组。各组术前连续灌胃7 d,于末次给药30 min后造模,采用胸膜腔注射角叉菜胶诱导胸膜炎模型。造模8 h后检测胸腔渗出液体积、渗出液中白细胞计数、蛋白含量、前列腺素E_2(PGE_2)、丙二醛(MDA)的含量及超氧化物歧化酶(SOD)、髓过氧化物酶(MPO)的活性。结果与正常组比较,模型组大鼠胸腔渗液体积和白细胞总数显著升高(P<0.01)。与模型组比较,YLS水提取物及其总黄酮能够减少大鼠胸腔渗出液体积、白细胞数量、蛋白含量、PGE_2及MDA的含量并降低MPO活性,提高胸腔渗出液SOD活性。结论YLS提取物对胸膜炎有明显的抑制作用,可能是通过抑制炎症介质PGE_2的生成、抑制白细胞浸润和游走及抗氧自由基等环节产生抗炎作用的。     

Changes of phospholipase D activity of rat peritoneal mast cells in degranulation

IM: To study the changes of phospholipase D (PLD) activity of actively sensitized rat peritoneal mast cells (RPMC) in degranulation. METHODS: Degranulation of RPMC was determined by measurement of β-hexosaminidase release. PLD activity assay was carried out by measurement of PLD product, choline, with chemiluminescent oxidation of luminol. RESULTS: Actively sensitized RPMC challenged with ovalbumin (0.5-8 mg/L for 120 s, 4 mg/L for 15-120 s) resulted in significant activation of PLD accompanied with the increment of P-hexosaminidase release. PLD activity of sensitized RPMC was increased by more than 2-fold compared with that of unsensitized RPMC which contained low levels of PLD activity [(35±13) pmol choline/min in 1×106cells], but β-hexosaminidase releases of the sensitized cells were as low as spontaneous releases.     

Local anti-inflammatory activity of steroid-21-oate esters in the carrageenan pleurisy model of acute inflammation

Local anti-inflammatory activities in the carrageenan (CGN)-induced pleurisy model are reported for steroid-21-oate esters derived from prednisolone. Following administration of equipotent doses (per rat) of prednisolone (P),.5mg; methyl 17,20α-acetonidodihydroprednisolonate (P4AC), 1 mg; methyl 20β-dihydroprednisolonate (P4B), 1.3 mg; methyl 20α-dihydroprednisolonate (P4A), 5.8 mg; and methyl 17,20β-acetonidodihydroprednisolonate (P4BC), 6 mg, effects of the steroids on exudate volume, leukocyte recruitment, and enzyme levels of cell-free exudates and washed exudate cells were assessed. All derivatives tested significantly inhibited emigration of neutrophils (PMNs) and monocytes (MNs). Steroid treatment with the β-isomers reduced exudate volumes by 41% and 56% for P4B and P4BC, respectively. In 5 hr neutrophilic exudates, free β-glucuronidase levels were reduced by 43% by either P4B or P4AC treatments. Of the derivatives tested P4B and P4AC treatments resulted in the greatest reduction of free lysozyme levels. Activities of membrane-bound γ-glutamyl-transferase (GGTP) in exudate cell pellets were reduced following treatment with P and the 20 alpha-epimers P4AC and P4A. Systemic effects assessed as decreases in plasma corticosterone or suppression of circulating lymphocytes were noted only following P treatment. These results suggest that 1) the locally active steroid-21-oate esters act at the site of inflammation by inhibiting PMN and MN infiltration in a manner congruous with conventional steroids, 2) steroid stereochemical configuration plays a role in effects on exudate volumes and GGTP activities, and 3) due to the absence of systemic effects, the steroid-21-oate esters may be safer local anti-inflammatory agents.     

Effect of capsaicin on carrageenan-induced inflammation in rat pleurisy and exudate substance P level.

Injection of 2.5 mg of lambda-carrageenan into the rat pleural cavity resulted in a time-dependent increase in pleural exudate substance P (SP) levels up to 24 hr. Synergistic increases in the exudate formation were observed when a sub-optimal quantity of carrageenan was injected with SP. Pre-treatment of rats with capsaicin at 50 and 100 mg/kg s.c. daily for one week prior to the induction of pleurisy blocked the increase in exudate volume and SP levels when compared to that normally detected after carrageenan injection. These results suggest that inhibition of SP production may improve inflammatory conditions.     

粉防己碱的抗炎作用与磷脂酶A_2的关系

为探讨Tet的抗炎作用及机制,在大鼠胸膜腔内注射Car复制胸膜炎。注射后2—48h,Neu-和ACC-PLA_2活性明显增强,胸膜渗出液量、蛋白质渗出量和白细胞游出数随之增加。Tet剂量依赖性地抑制上述各指标的变化,且Tet引起的PLA_2活性的降低与炎症指标的减少密切相关。提示Tet有良好抗炎作用,其机制可能涉及抑制PLA_2激活和释放。     

Modulation of exudate inflammation parameters in rat carrageenan-induced granuloma by modification of exudate iron levels

We have used the carrageenan-induced pouch-granuloma in rats to investigate how changes in low-molecular-mass iron chelate levels in the exudate, induced by iron loading (iron-dextran) or chelation (desferrioxamine) influence cellular and systemic inflammatory parameters. In the iron-treated group we observed a rapid decrease in the number of leukocytes and exudate volume; there was also an increase in ferritin iron and low-molecular-mass iron chelates, and on the eighth day a systemic response. In the desferrioxamine-treated group we detected a decrease in low-molecular-mass iron chelates, ferritin iron, and an increase in the number of leukocytes. We describe the protective effects of desferrioxamine against the deleterious effects of ferrous iron and relate this to its chelating and scavenging activity. The results suggest that the levels of low-molecular-mass iron chelates modulate the inflammatory response, possibly through their contribution to the oxygen free radical generation, which is responsible for the cell membrane damage and subsequently its death. The modulatory action of iron-dextran and desferrioxamine support our hypothesis.