Age-related changes in seminal polymorphonuclear elastase in men with asymptomatic inflammation of the genital tract

Aim： To investigate age-related inflammatory events in the male genital tract. Methods： In a total of 4 265 randomly collected patients attending the andrological outpatient clinic of the Center for Dermatology and Andrology, University of Giessen, Germany, ejaculate volume, pH-value, sperm concentration, total and progressive sperm motility, concentration of polymorphonuclear （PMN） elastase, number of peroxidase-positive cells and fructose were measured and correlated with patient＇s age. Results： While ejaculate volume, motility and fructose all correlated negatively with age, sperm concentration, PMN elastase and the pH-value showed a positive correlation. The prevalence of male genital tract inflammation （as defined by PMN elastase 〉 250 ng/mL） and its severity increased significantly. PMN elastase did not correlate with sperm motility. Fructose as a marker of seminal vesicle function showed a significant negative relationship with the PMN elastase levels, the number of peroxidase-positive cells and sperm motility. Conclusion： The significant increases of PMN elastase levels as marker of male genital tract inflammation in older men appear to be indicative of age-related changes in local immunoregulatory mechanisms. Because there is no association of PMN elastase with sperm motility, a direct inhibitory effect of this enzyme can be excluded.

Measurement of calprotectin in ascitic fluid to identify elevated polymorphonuclear cell count

AIM: To evaluate the diagnostic capability of calprotectin in ascitic fluid for detecting a polymorphonuclear (PMN) cell count > 250/μL ascites. METHODS: In this prospective observational study, a total of 130 ascites samples were analysed from 71 consecutive patients referred for paracentesis. Total and differential leukocyte cell counts were determined manually with a Neubauer chamber and gentianviolet stain. Calprotectin was measured in 1 mL ascetic fluid by enzyme-linked immunosorbent assay (ELISA) and a point-of-care (POC) lateral flow assay with the Quantum Blue Reader (Bühlmann Laboratories). All measurements were carried out in a central laboratory by senior personnel blinded to patient history. A PMN count > 250/μL was the primary endpoint of the study. The diagnostic value of ascitic calprotectin measurement was assessed by comparing to the final diagnosis of each patient that had been adjudicated by investigators blinded to calprotectin values. RESULTS: The PMN count was > 250/μL in 19 samples (14.6%) from 15 patients (21.1%) and varied widely among the study population (range 10-19 800/mL and 1-17 820/mL, respectively). Spontaneous bacterial peritonitis (SBP) was the final diagnosis in four patients (5.6%). All patients with PMN ≤ 250/μL had negative bacterial culture. PMN count was elevated in five patients with peritoneal carcinomatosis, three with lymphoma, one with neuroendocrine carcinoma, and two with secondary peritonitis due to abdominal perforation. PMN cell counts correlated with ascitic calprotectin values (Spearman's rho; r = 0.457 for ELISA, r = 0.473 for POC). A considerable range of ascitic calprotectin concentrations was detected by ELISA [median 0.43 μg/mL, interquartile range (IQR) 0.23-1.23 (range 0.10-14.93)] and POC [median 0.38 μg/mL, IQR 0.38-0.56 (range 0.38-13.31)]. Ascitic calprotectin levels were higher in samples with PMN > 250/μL, by both ELISA [median (IQR) 2.48 μg/mL (1.61-3.65) vs 0.10 μg/mL (0.10-0.36), P < 0.001] and POC [2.78 μg/mL (2.05-5.37) vs 0.38 μg/mL (0.38-0.41), P < 0.001]. The area under the receiver operating characteristics curve for identifying an elevated PMN count was 0.977 (95%CI: 0.933 to 0.995) for ELISA and 0.982 (95%CI: 0.942 to 0.997) for POC (P = 0.246 vs ELISA). Using the optimal cut-off value for ELISA (0.63 μg/mL), ascitic calprotectin had 94.8% sensitivity, 89.2% specificity, positive and negative likelihood ratios of 8.76 and 0.06 respectively, positive and negative predictive values of 60.0% and 99.0% respectively, and 90.0% overall accuracy. Using the optimal cut-off value for POC (0.51 μg/mL), the respective values were 100.0%, 84.7%, 6.53, 0.00, 52.8%, 100% and 87.7%. Correlation between ELISA and POC was excellent (r = 0.873, P < 0.001). The mean ± SD of the difference was -0.11 ± 0.48 μg/mL with limits of agreement of + 0.8 μg/mL (95%CI: 0.69 to 0.98) and -1.1 μg/mL (95%CI: -1.19 to -0.91). CONCLUSION: Ascitic calprotectin reliably predicts PMN count > 250/μL, which may prove useful in the diagnosis of SBP, especially with a readily available bedside testing device.

Protective Effect of Distillate and Redistillate of Cow’s Urine in Human Polymorphonuclear Leukocytes Challenged With Established Genotoxic Chemicals

Objective From the ancient period cow’urine has been used as a medicine. In Veda, cow’urine was compared to the nectar. In Susrut, several medicinal properties of cow’ urine have been mentioned and are known to cause weight loss, reversal of certain cardiac and kidney problems, indigestion, stomach ache, edema, etc. However, the literature and scripture did not mention the antigenotoxic properties of cow’urine. Methods In the present investigation, the antigenotoxic/ antioxidant properties of cow’ urine distillate and redistillate were studied in vitro. The antioxidant status and volatile fatty acid levels were determined. Actinomycin-D (0.1ol/L) and hydrogen peroxide (150 mol/L) were used for inducing DNA strand break with 0.1% DMSO as negative control. Dose for the antigenotoxic effect of cow’ urine was chosen from the dose response study carried out earlier. Results Both actinomycin-D and H2O2 caused statistically significant DNA unwinding of 80% & 75% respectively (P<0.001) as revealed by fluorimetric analysis of DNA unwinding (FADU), and the damage could be protected with the redistilled cow urine distillate (1, 50 & 100 ) in simultaneous treatment with genotoxic chemicals. Conclusion The redistillate of cowurine was found to possess total antioxidant status of around 2.6 mmol, contributed mainly by volatile fatty acids (1500 mg/L) as revealed by the GC-MS studies. These fatty acids and other antioxidants might cause the observed protective effects.

Study of plasma effects during hypoxia and hemorrhagic shock on polymorphonuclear neutrophil-vascular endothelial cell interactions in vitro

The plasma effects during hypoxia and hemorrhagic shock on the interactions between plymorphonuclear neutrophils (PMNs) and cultured pulmonary artery endothelial cells (PAECs) were studied. The plasma samples were obtained from the goats under the following conditions: (1)Normal control plasma was obtained from the goats at sea level to aserve as the control (CP). (2)Hypoxic plasma was obtained after the goats were exposed to a simulated altitude of 4 000 m for 24 h (HP). (3) Hypotensive hypoxic plasma was obtained after the goats were bled to a mean arterial pressure of 5. 5±0. 3 kpa for 1 h under hypoxic condition (HHP). (4) Retransfused hypoxic plasma was obtained when the hypotensive goats were transfused with the shed blood for 4 h under hypoxic condition (RHP). It was found that HP , HHP and RHP especially RHP exerted profound effects on the activities of PMNs and PAECs in a concentration and time dependent manner after the PMNs and PAECs were incubated in the media containing different concentrations of the 4 kinds of plasma for different durations. Low concentration of RHP (less than 12. 5%) significantly increased the activity of PAECs (P<0. 01 vs CP) but its high concentration (more than 12. 5%) markedly decreased their activity (P<0. 01 vs CP, HP and HHP). HP, HHP and RHP increased the activity of PAECs in the early stage of incubation (1 to 3 h) (P<0. 01 vs CP) but decreased it in the late stage (6 to 12 h) (P<0. 01 vs CP). The activity of PMNs was significantly increased after 1 h incubation with HP, HHP and RHP (PM0. 001) and this effect was also concentrationdependent.The effects of RHP was the most potent, HHP the next and HP the least. The deformability of PMNs was significantly decreased (P <0. 001) after they were incubated in RHP for 3 h. The adhesive force of PMNs and PAECs was also significantly increased after 12 h incubation with RHP. These findings suggest that there are substances in the hypoxic plasma to activate or damage the interactions between PMNs and PAECs and the amounts of the substances are further increased in hypotensive hypoxic plasma and retransfused hypoxic plasma and the 'activation-damage'to PMNs and PAECs and the subsequent interactions between PMNs and PAECs play an important role in the pathological changes of hypoxia and hemorrhagic shock.

Serum Resistin Level and Polymorphonuclear Leukocytes Dysfunctions in Children on Regular Hemodialysis

Resistin is a secretory adipocytoine, which is expressed mainly in humans by inflammatory cells especially macrophages. Resistin serum levels are elevated in end-stage renal diseases of people having an increased risk of infections as a result of impaired polymorphonuclear leukocytes (PMNLs) functions. Objectives: To evaluate neutrophil functions (phagocytosis and oxidative burst) in children with end-stage renal disease (ESRD) on regular hemodialysis and to shed light on the contribution of resistin on neutrophil functions. Patients and Methods: The study included 40 children with ESRD on regular hemodialysis. Their ages ranged from 6 to 12 years, and they were selected from children attending the pediatric hemodialysis unit of AL-Zahraa Hospital, Al-Azher University during the period from October 2012 to December 2013. Another group of 40 apparently healthy children with matched age and sex with the patient group served as a control. Serum resistin, phagocytic index and nitro blue tetrazolium test (NBT%) were assessed in both groups. Results: There was a statistically more significant increase in resistin serum levels in cases than in controls;it was (3.25 ± 0.86 ng/ml) and (0.25 ± 0.16 ng/ml) respectively

ANTAGONISTIC EFFECTS OF GRANULOCYTE DERIVED INHIBITORS AND CYTOKINES ON APOPTOSIS IN POLYMORPHONUCLEAR LEUKOCYTES

An extract (G-INH) made from mature human granulocytes freshly isolated from normai blood causes human neutrophils to undergo apoptosis in vitro as shown by morphologic changes and by the typical ladder pattern of small DNA fragments noted on agarose gel electrophoresis of isolated DNA. Apoptosis occurs in from 20% to 30% of neutrophils over 24 hours of culture in vitro and the addition of G-INH to the medium causes a dose-related increase in the incidence of apoptosis. Heating G-INH at 60t for 30 minutes does not destroy its capacity to induce apoptosis but GM-CSF, G-CSF, and to a lesser extent IL-1β, antagonize this action. IL-3 does not diminish G-INH induced apoptosis of neutrophils. Substances, released from, mature neutrophils may participate in regulating the survival of other neutrophils, particularly in sites where the cells are in close proximity as in the marrow. Self destruction of post-mitotic neutrophils in marrow may thus represent an-other level at which regulation of cell production

Molecular basis of the adhesive process of polymorphonuclear neutrophils to vascular endothelial cells under endotoxin condition

The monocellular adhesive ability of polymorphonuclear neutrophils (PMN)to vascular endothelial cells (VEC) was observed with micromanipulation technique when the cells were treated with endotoxin, Anti-CD18 monoclonal antibodies (McAb),anti-endothelial-leucocytic adhesion molecule-l (ELAM-l)McAb and anti-intercellular adhesion molecule-1 (ICAM-1) McAb and the effects of these 3 adhesion molecules on the development of adhesion force between PMNs and VECs were investigated. It was found that CD18 McAb decreased obviously the adhesive ability of PMNs in the whole course of adhesion increasing and the McAbs of ELAM-1 and ICAMI-1 inhibited most of adhesive force of VECs in the 4th and 12th hour of the course respectively.These findings suggest that the main molecular basis of rapid increase of adhesive ability of PMNs is CD18 expression and that of delayed increase of adhesive ability of VECs is ELAM-1 and ICAM-1 expression.

The Observation of Complement Activation and Polymorphonuclear Neutrocytopenia during Cardiopulmonary Bypass

By determining the plasma levels of C3, C4, factor B and polymorphonuclear neutrophils (PMNs) of the patients who received CPB, the path of complement activation and changes of PMNs were studied. The results suggest that complement system was activated through alternative pathway during CPB and was activated through classic pathway after CPB. The anaphylatoxin, the products of complement activation may be responsible for the polymorphonuclear neutrocytopenia.

The effects of amniotic membrane on polymorphonuclear cells

Objective To investigate the effects of fresh and preserved amniotic membrane on polymorphonuclear neutrophils (PMNs) so as to understand the anti-inflammatory mechanism of amniotic membrane transplantation.Methods Conditioned medium was collected 48 hours after fresh or preserved amnions were cultured in DMEM and 5% CO 2 at 37℃. Then, polymorphonuclear cells were cultured in conditioned culture or DMEM. Fluorescent microscopy with 4’,6-diamidino-2-phenylindole (DAPI) staining and cytometry were performed 6, 9, 12, and 15 hours later. Results Apoptotic neutrophils were found in each group at different time points. The percentage of apoptotic cells at 6, 9, 12, and 15 hours after culture in the fresh and preserved amnion groups and the control group was 17.3%, 24.4%, 29.8%, 37.1%, and 16.2%, 20.1%, 23.7%, 27.7%, and 10.2%, 13.7%, 21.1%, 26.4%, respectively (t test, P 1<0.01, P 2<0.01 and P 3<0.01).Conclusion Amniotic membrane can accelerate apoptosis of polymorphonuclear neutrophils, reduce inflammation, and prevent ocular surface collagen from resolution, indicating that fresh amnion might have a stronger effect than preserved amnion.

Continuous activation of polymorphonuclear myeloid-derived suppressor cells during pregnancy is critical for fetal development

The maternal immune system is vital in maintaining immunotolerance to the semiallogeneic fetus for a successful pregnancy.Although studies have shown that myeloid-derived suppressor cells(MDSCs)play an important role in maintaining feto-maternal tolerance,little is known about the role of MDSCs in pregnancies with intrauterine growth retardation(IUGR).Here,we reported that the activation of polymorphonuclear myeloid-derived suppressor cells(PMN-MDSCs)during pregnancy was closely associated with fetal growth.In humans,class E scavenger receptor 1(SR-E1),a distinct marker for human PMN-MDSCs,was used to investigate PMN-MDSC function during pregnancy.Continuous activation of SR-E1+PMN-MDSCs was observed in all stages of pregnancy,accompanied by high cellular levels of ROS and arginase-1 activity,mediated through STAT6 signaling.However,SR-E1+PMN-MDSCs in pregnancies with IUGR showed significantly lower suppressive activity,lower arginase-1 activity and ROS levels,and decreased STAT6 phosphorylation level,which were accompanied by an increase in inflammatory factors,compared with those in normal pregnancies.Moreover,the population of SR-E1+PMN-MDSCs was negatively correlated with the adverse outcomes of newborns from pregnancies with IUGR.In mice,decreases in cell population,suppressive activity,target expression levels,and STAT6 phosphorylation levels were also observed in the pregnancies with IUGR compared with the normal pregnancies,which were rescued by the adoptive transfer of PMN-MDSCs from pregnant mice.Interestingly,the growth-promoting factors(GPFs)secreted by placental PMN-MDSCs in both humans and mice play a vital role in fetal development.These findings collectively support that PMN-MDSCs have another new role in pregnancy,which can improve adverse neonatal outcomes.

APE1通过免疫抑制性肿瘤微环境介导结肠炎相关结直肠癌的发生发展

目的探讨脱嘌呤/脱嘧啶核酸内切酶1(apurinic/apyrimidinic endonuclease 1,APE1)在慢性肠道炎症向结肠炎相关性结直肠癌(colitis-associated colorectal cancer,CAC)转化过程中的调控机制。方法将C64S点突变纯合子(APE1^(C64S))和野生型(APE1^(WT))小鼠分别按随机数字表法分为实验组与对照组,实验组应用氧化偶氮甲烷(azoxymethane,AOM)及葡聚糖硫酸钠盐(dextran sulfate sodium salt,DSS)溶液构建CAC体内模型,采用免疫组化及多重免疫荧光分析各组小鼠结肠组织APE1表达及免疫细胞浸润情况。通过慢病毒转染构建APE1稳定敲低的小鼠结肠癌MC38细胞系,并对APE1^(WT)与APE1^(C64S)小鼠进行皮下荷瘤实验以确定肿瘤细胞来源的APE1导致免疫抑制性肿瘤微环境,采用免疫组化及多重免疫荧光分析荷瘤标本APE1、趋化因子(C-X-C基序)配体1[chemokine(C-X-C motif)ligand 1,CXCL1]表达及免疫细胞浸润情况。对来自陆军特色医学中心的1名28岁女性CAC患者的肿瘤标本采用免疫组化及多重免疫荧光分析肿瘤及邻近炎症组织中APE1、CXCL1的表达及免疫细胞浸润情况。结果与对照组及APE1^(WT)实验组相比,APE1^(C64S)实验组小鼠的疾病活动指数及肿瘤形成数量明显降低,多形核髓源抑制细胞(polymorphonuclear myeloid-derived suppressor cells,PMN-MDSCs)浸润显著减少,CD4^(+)及CD8^(+)T细胞显著增多(P<0.05)。APE1^(WT)与APE1^(C64S)皮下荷瘤小鼠肿瘤生长及各免疫细胞差异无统计学意义;而使用敲低APE1的肿瘤细胞进行荷瘤实验,发现肿瘤生长明显抑制,PMN-MDSCs浸润减少,同时CD4^(+)及CD8^(+)T细胞显著增多(P<0.05)。在CAC患者肿瘤组织中APE1高表达、PMN-MDSCs浸润增加,CD8^(+)T细胞在肿瘤组织中较炎症组织浸润显著减少(P<0.05)。结论肿瘤细胞中APE1的氧化还原功能可促进PMN-MDSCs肿瘤浸润,同时减少T细胞的数量,从而形成免疫抑制性肿瘤微环境介导CAC的发生发展。

痰中性粒细胞数量和结核分枝杆菌载量预测肺结核患者强化治疗的反应

目的探讨痰涂片阳性肺结核(PTB)患者中痰中性粒细胞(PMNs)数量、结核分枝杆菌载量与抗结核强化治疗反应的关系。方法回顾性收集2021年1—12月陕西省结核病防治院患者,以痰未转阴组为病例组,痰转阴组为对照组,分析新发PTB患者痰PMNs数量、结核分枝杆菌载量和其他因素的治疗前后水平。结果与痰未转阴组比较,痰转阴组患者治疗前PMNs数量、显微镜下结核分枝杆菌以及痰涂片抗酸杆菌(AFB)镜检分级、痰GeneXpert测定的载量更低,而痰GeneXpert测定的Probe B循环阈值(Xpert-Probe B-Ct)更高(P<0.05)。Xpert-Probe B-Ct值与AFB显微镜测定的相应痰样本中的结核分枝杆菌载量显著相关(rs=-0.77,P<0.001)。在受试者操作特征曲线分析中,痰PMNs对痰转阴延迟的预测效能最高,Xpert-Probe B-Ct值次之,镜检下痰结核分枝杆菌载量最差,曲线下面积分别为0.852、0.784、0.669。根据多因素回归模型校正,痰PMNs数量、GeneXpert测定的痰结核分枝杆菌载量可能是影响临床抗结核治疗反应的主要因素(P<0.05)。结论治疗前痰PMNs数量和GeneXpert测定的痰结核分枝杆菌载量具有成为预测PTB患者强化治疗后治疗反应标志物的潜力。

人脐带间充质干细胞移植治疗角膜碱烧伤的实验研究

目的:研究人脐带间充质干细胞(hUCMSCs)移植治疗兔角膜碱烧伤的疗效,分析多形核中性白细胞(PMNs)的浸润和血管内皮生长因子(VEGF)的表达变化。方法:将75只健康日本白兔随机分成A、B、C组,每组25只,均建立右眼角膜碱烧伤模型。A组兔右眼角膜碱烧伤后立即行角膜病灶区羊膜负载hUCMSCs覆盖术;B组兔右眼角膜碱烧伤后立即行角膜病灶区羊膜覆盖术;C组兔右眼角膜碱烧伤后未进行处理。角膜碱烧伤后3、7、14、21、28d,裂隙灯下观察实验兔角膜恢复情况并照相,对角膜新生血管(CNV)生长情况进行评分,并分离角膜组织制作病理切片,通过苏木素-伊红(HE)染色观察PMNs浸润情况,采用免疫组化染色法测定VEGF的表达。结果:角膜碱烧伤后14d时,A组CNV较B组生长明显缓慢,A组病灶区周围CNV生长评分明显低于B组(P<0.05)。碱烧伤后3d时,角膜PMNs数量增加,浸润角膜基质层,7d时稍有下降,14d时达到峰值,后渐进性下降,碱烧伤后早期浸润在病灶区角膜基质内,后期浸润范围与溃疡面积相同,碱烧伤后各时间点A组和B组角膜PMNs密度均明显低于C组(P<0.05),且14、21d时,A组均明显低于B组(P<0.05)。碱烧伤后各组角膜VEGF表达水平均在7~14d时达到峰值,28d时明显降低,碱烧伤后各时间点A组和B组VEGF表达水平均明显低于C组(P<0.05),且7、14、21d时,A组均明显低于B组(P<0.05)。结论:羊膜负载hUCMSCs移植治疗兔角膜碱烧伤可减少CNV形成,抑制碱烧伤后角膜血管化,角膜病理损伤及血管化与PMNs和VEGF密切相关。

PMN-MDSCs细胞对异基因造血干细胞移植术后患者T淋巴细胞增殖及干扰素-γ分泌的影响

目的探讨多形核的髓系来源抑制性细胞(PMN-MDSCs)对异基因造血干细胞移植(allo-HSCT)术后患者T淋巴细胞增殖和干扰素-γ(IFN-γ)分泌的影响。方法抽取16例allo-HSCT术后患者第80天及8例健康供者任意时间点空腹全血标本,先采用中性粒细胞分离液分离2组受检者外周血含PMN-MDSCs与T细胞的白细胞、再采用流式分选出患者PMN-MDSCs与T细胞及健康供者PMN-MDSCs;采用瑞氏-姬姆萨(WG)染色观察2组受检者外周血PMN-MDSCs的形态学特征,Annexin V-APC/7-AAD法检测0 h、12 h、24 h及48 h时allo-HSCT术后患者外周血PMN-MDSCs细胞凋亡情况,羧基荧光素琥珀酰亚胺酯(CFSE)染色和荧光酶联免疫斑点(ELISpot)法检测allo-HSCT术后患者外周血PMN-MDSCs对T细胞增殖及IFN-γ表达的影响。结果allo-HSCT术后患者PMN-MDSCs多呈不成熟粒细胞表型,健康供者PMN-MDSCs呈成熟粒细胞表型,且患者PMN-MDSCs于48 h时大部分仍处于存活状态;CFSE和ELISpot实验表明allo-HSCT术后患者PMN-MDSCs可抑制其T淋巴细胞的增殖、分裂及胞内IFN-γ的释放。结论PMN-MDSCs可抑制allo-HSCT术后患者T淋巴细胞的增殖及分泌INF-γ。

外周血中性粒细胞和口腔中性粒细胞免疫炎症反应的差异比较

目的:比较口腔中性粒细胞(oPMN)和外周血中性粒细胞(cPMN)免疫炎症反应的差异,探讨oPMN维持牙周健康的作用。方法:常规密度梯度离心法分离获得健康人外周静脉血来源的cPMN,离心过滤法分离获得健康人口腔含漱液来源的oPMN。采用1μg/mL牙龈卟啉单胞菌(P.gingivalis)脂多糖(LPS)刺激,并以大肠杆菌(E.coli)LPS作为阳性对照,流式细胞术检测凋亡水平变化;Transwell实验比较白介素(IL)-8刺激前后两种细胞趋化能力的差异;ELISA检测IL-1β、IL-8和IL-10表达水平变化;流式细胞术检测两种细胞对P.gingivalis或E.coli的吞噬作用;免疫荧光实验观察中性粒细胞胞外诱捕网(NETs)的形成,Sytox Green法对胞外DNA进行定量;Western blot检测丝裂原活化蛋白激酶1/2(MEK1/2)、细胞外信号调节蛋白激酶1/2(ERK1/2)和核转录因子-κB(NF-κB)P65磷酸化水平的变化。结果:两种LPS刺激后,与cPMN相比,oPMN凋亡增多(P<0.05)。IL-8刺激后,两种细胞趋化能力未见明显差异(P>0.05)。两种LPS刺激后,oPMN较cPMN的IL-1β、IL-8和IL-10表达水平更高(P<0.01)。oPMN吞噬P.gingivalis和E.coli的能力强于cPMN(P<0.05),形成的NETs更多,胞外DNA水平、MEK1/2、ERK1/2和NF-κB P65磷酸化水平均较cPMN更高(P<0.05)。结论:oPMN可能是具有过度活跃表型的中性粒细胞,其吞噬、分泌细胞因子和形成NETs的能力较cPMN增强,可能与其MEK1/2、ERK1/2和NF-κB磷酸化水平增高有关。

原花青素抗促癌物诱发H_2O_2释放及脂质过氧化

目的 探讨葡萄原花青素抗氧化和肿瘤化学预防作用机制。方法 以大鼠多形核白细胞 (PMNs)为材料 ,利用酚红氧化原理比色测定了原花青素对巴豆油 (crotonoil)刺激PMNs生成H2 O2 的影响。结果 原花青素能显著性抑制巴豆油刺激PMNs生成H2 O2 ;以原花青素的大鼠含药血清代替反应系统中的药物 ,同样观察到原花青素具有抑制H2 O2 释放的作用 ,该作用在给药后 1h左右最强 ,且具有一定的时效关系和量效关系。对巴豆油诱发的小鼠肝线粒体脂质过氧化 ,原花青素具明显抑制作用 ,能明显提高肝线粒体SOD活力 ,减少MDA生成。

羟基红花黄色素A对血小板活化因子的拮抗作用

目的 观察羟基红花黄色素A(HSYA)对血小板活化因子 (PAF)的拮抗作用。方法 以放射受体结合试验观察HSYA抑制 [3 H]PAF与家兔洗涤血小板 (WRP)特异性结合的作用 ,以比浊法观察HSYA抑制PAF介导的WRP及兔多型核白细胞 (PMNs)聚集的作用。结果 HSYA可浓度依赖地抑制 1 ,2及 4nmol·L-1 [3 H]PAF与WRP受体的结合 ;HSYA抑制PAF介导的WRP及兔PMNs聚集 ,均具明显的量效关系 ,其IC50 分别为 0 99及 0 70mmol·L-1 。

麝香糖蛋白成分对中性白细胞趋化反应的抑制作用

目的 :观察麝香糖蛋白成分 (麝香 1)对中性白细胞趋化反应的影响。方法 :体内采用羧甲基纤维素诱导大鼠腹腔内中性白细胞趋化反应模型。体外采用Boyden隔室法测定兔中性白细胞趋化反应。以Fura 2标记荧光法测定细胞内游离钙水平。结果 :皮下注射麝香水提物 5 ,2 0 ,80mg·kg-1,显著抑制羧甲基纤维素诱导的大鼠腹腔中性白细胞趋化反应 ,抑制率分别为 5 8.6 % ,6 5 .1%和 73 .0 % ;终浓度为 1,10 ,10 0 μg·mL-1麝香 1,显著抑制fMLPP诱导的兔中性白细胞的趋化反应 ,抑制率分别为 5 7.9% ,6 4.7%和 77.8% ;终浓度为 1,10 ,10 0 μg·mL-1的麝香 1,明显抑制刺激剂引起的中性白细胞胞内游离钙水平的升高 ,抑制率分别为 16 .3% ,2 4.7%和 37.8%。结论 :麝香糖蛋白成分对中性白细胞趋化反应有显著抑制作用 ,是其抗炎作用机制之一。

黄芪的三种提取成分对氧自由基作用的影响

研究从黄芪中分离出的总黄酮（ＴＦＡ）、总皂甙（ＴＳＡ）和总多糖（ＴＰＡ）对黄嘌呤／黄嘌呤氧化酶和多形核白细胞呼吸爆发介导鲁米诺化学发光及Ｈ２Ｏ２Ｆｅ２＋诱导鼠肝匀浆脂质过氧化三个实验体系的作用。结果表明，ＴＦＡ和ＴＳＡ有良好的清除氧自由基作用，其中ＴＦＡ效果最好，ＴＳＡ次之，提示黄芪抗氧化作用的主要成分可能是ＴＦＡ和ＴＳＡ。ＴＰＡ对氧自由基有部分清除作用．但作用强度随其深度的增加而降低。

银杏叶的化学成分及其抗氧化活性

目的:研究银杏叶中化学成分的抗氧化活性。方法:对银杏叶提取物进行化学成分的分离鉴定,采用化学发光法,测定所得成分对邻苯三酚-鲁米诺-碳酸缓冲液体系产生的超氧阴离子(O.2-)的清除能力,以及对大鼠中性粒细胞(PMN)呼吸爆发的抑制作用。结果:分得9种黄酮类成分及4种萜内酯类成分,分别为:槲皮素(Ⅰ)、山柰酚(Ⅱ)、异鼠李素(Ⅲ)、木犀草素(Ⅳ)、异银杏素(Ⅴ)、槲皮苷(Ⅵ)、芦丁(Ⅶ)、槲皮素-3-O-(2″-O-(6-对羟基-反式-桂皮酰)-β-D-葡萄糖)-α-L-鼠李糖苷(Ⅷ)、苜蓿草素-7-O-β-D-葡萄糖苷(Ⅸ)、银杏内酯A(Ⅹ)、银杏内酯B(Ⅺ)、银杏内酯C(Ⅻ)、白果内酯(XⅢ),其中化合物Ⅸ为首次从银杏叶中分离得到。黄酮类化合物均具有不同程度的清除O2.-及抑制PMN呼吸爆发作用,活性对结构依赖性明显;内酯类成分不能清除自由基,仅对PMN呼吸爆发有微弱抑制作用。结论:银杏叶体外抗氧化作用的主要活性成分为黄酮类成分,内酯类成分无明显抗氧化作用。