Samsum ant venom modulates the immune response and redox status at the acute toxic dose in vivo

Background:Ant venoms express surface molecules that participate in antigen presentation involving pro- and anti-inflammatory cytokines. This work aims to investigate the expression of MHC-II, CD80 and CD86 on the polymorphonuclear cells (PMNs) in rats injected with samsum ant venom (SAV).Methods:Rats were divided into three groups - control, SAV-treated (intraperitoneal route, 600 μg/kg), and SAV-treated (subcutaneous route, 600 μg/kg). After five doses, animals were euthanized and samples collected for analysis.Results:The subcutaneous SAV-trated rats presented decreased levels of glutathione with increased cholesterol and triglyceride levels. Intraperitoneal SAV-treated animals displayed significantly reduced concentrations of both IFN-γ and IL-17 in comparison with the control group. However, intraperitoneal and subcutaneous SAV-treated rats were able to upregulate the expressions of MHC-II, CD80 and CD86 on PMNs in comparison with the control respectively. The histological examination showed severe lymphocyte depletion in the splenic white pulp of the intraperitoneal SAV-injected rats.Conclusion:Stimulation of PMNs by SAV leads to upregulation of MHC-II, CD 80, and CD 86, which plays critical roles in antigen presentation and consequently proliferation of T-cells. Subcutaneous route was more efficient than intraperitoneal by elevating MHC-II, CD80 and CD86 expression, disturbing oxidative stability and increasing lipogram concentration.

The effect of concentration ratio on tumor cell adhesion in shear flow / 医用生物力学

Objective To study the different effects from different concentration ratios of polymorphonuclear neutrophil (PMN) to tumor cell (TC) on the process of tumor cell adhesion to endothelial cell (EC) in shear flow. Methods PMNs and TCs with different concentration ratios (PMN-TC ratio) were added into the parallel plate flow chamber, and changes in the numbers of transient and accumulative adhered TCs on ECs at different shear rates (50 s-1，100 s-1，200 s-1) were analyzed. Results The transient and accumulative adhesion of TCs on ECs at PMN-TC ratio of 3︰1 significantly increased as compared to that at PMN-TC ratio of 1︰1, especially under high shear flow condition (100 s-1 and 200 s-1). Moreover, in the 5 minute-observation period, the effect of PMN-TC ratio on TC adhered to ECs occurred earlier when the shear rate increased. Conclusions The increase of PMN-TC concentration ratio can promote TC adhesion to ECs in shear flow, and the research findings provide significant references for studying TC metastasis in blood vessels and the target therapy of tumors.

Does Propofol Improve Postischemic Myocardial Dysfunction by Reducing the Postischemic Adhesion of Polymorphonuclear Neutrophils? / 대한마취과학회지

BACKGROUND: Adhesion of polymorphonuclear neutrophils (PMNs) to the coronary vascular endothelium is a crucial step in the development of postischemic myocardial injury. Propofol, a free radical scavenger has been demonstrated to provide cardioprotective effects by attenuating ischemia and reperfusion injury. The purpose of this study was to investigate whether propofol inhibits the postischemic coronary vascular adhesion of PMNs and results in a reduction of postischemic myocardial dysfunction in isolated guinea pig hearts. METHODS: 42 male guinea pigs were used in this study. Hearts were isolated and perfused with modified Krebs-Hanseleit solution. All isolated hearts were subjected in turn to stabilization for 10 min, perfusion for 15 min, global ischemia for 30 min and reperfusion for 60 min. The isolated hearts were then divided into 6 groups. Each group was subjected to different interventions as follow so. C group: No intervention was performed, except ischemia/reperfusion injury. PPF group: Propofol (4 microgram/ml) was infused from the start of perfusion to 5 min after the start of reperfusion. PMN group: PMNs were infused for 1 min on 2 minutes after reperfusion. P2PMN group: Propofol (2 microgram/ml) was infused from the start of perfusion to 5 min after the start of reperfusion, and PMNs were infused for 1 min on 2 minutes after reperfusion. P4PMN group: Propofol (4 microgram/ml) was infused from the start of perfusion to 5 min after the start of reperfusion, and PMNs were infused for 1 min on 2 minutes after reperfusion. IL-PMN group: Intralipid (0.4 mg/ml) was infused from the start of perfusion to 5 min after the start of reperfusion, and PMNs were infused for 1 min on 2 minutes after reperfusion. PMNs adhesion (%), left ventricular developed pressure (LVDP), dP/dt, heart rates and coronary flow were measured. RESULTS: Propofol (2 microgram/ml, 4 microgram/ml) significantly reduced postischemic PMNs adhesion compared with that of the PMN group (54.0+/-8.0%, 50.0+/-5.0% vs 72.5+/-6.5% P <0.05 respectively) and prevented the deterioration of myocardial function seen after PMNs application (LVDP 74.5+/-7.3%, 60.3+/-4.5% vs 48.4+/-2.7% respectively). CONCLUSIONS: These results show that propofol reduces the postischemic coronary vascular adhesion of PMNs and prevents postischemic myocardial dysfuncion.

E ffects of Vitamin E, Selenium and Zinc Sulfate on Superoxide Release from Human Polymorphonuclear Leuko cytes and Cultured Bovine Retinal Pigment Epithelial Cells

To evaluate antioxidative effect of vitamin E, selenium and zinc sulfate on release of oxygen free radicals from polymorphonuclear leukocytes[PMNs], I measured the amount of superoxide release from human PMNs stimulated by phorbol myristate acetate[PMA]with addition of some antioxidant and antioxidative micronutrients using superoxide-dependent cytochrome c reduction.And to determine a protective effect of them on cultured bovine retinal pigment epithelial cells [RPE]from oxygen radicals, I measured a viability of bovine RPE using MTT assay after incubation with human PMNs and PMA. Vitamin E, selenuim and zinc sulfate are utilized at different concentrations of 1, 10 and 20 microM. Vitamin E and zinc sulfate inhibited superoxide production from PMNs stimulated by PMA from 10 microM concentration gnificantly. But in case of selenium, significant antioxidative effect was not found at each concentration.The antioxidative effect on cultured bovine RPE was evaluated using MTT assay.Among antioxidant and antioxidative micronutrients tested, vitamin E and zinc sulfate inhibited free radical damage to bovine RPE, they increased cell survival rate on culture at concentration of 1, 10 and 20 microM. In contrast, selenium did not increase cell survival rate significantly. With these results, it was found that vitamin E and zinc sulfate had antioxidative effect against superoxide release from PMNs and also protective effect of bovine RPE from free radical damages.Recent studies supported that peroxide may play an important role in causing tissue damage in human and experimental models of ocular inflammation and possibly in Behcet`s disease.It was suggested that antioxidant vitamins and minerals like vitamin E or zinc sulfate might be useful for management and/or prevention of these conditions.

Effect of UV-B and Amniotic Membrane on Inflammation, Lipid Peroxidation and Keratocyte Apoptosis Induced by PRK

The authorsinvestigated the effects of UV-B and amniotic membrane graft about PRK induced inflammatory cell infiltration into corneal stroma, lipid peroxidation and keratocyte apoptosis. Total 20 white rabbits were divided into 5 groups; 1)mechanical epithelial removal, 2)epithelial removal and UV-B irradiation, 3)PRK only, 4) PRK and UV-B irradiation, 5)Amniotic membrane graft after PRK and UV-B irradiation. All corneas were harvested after 24hrs. H & E stain for PMNs infiltration, MDA immunohistochemical stain for lipid peroxidation and TUNEL stain for keratocyte apoptosis were performed. UV-B had little effect on infiltration of inflammatory cell into corneal stroma, lipid peroxidation and keratocyte apoptosis. Amniotic membrane suppressed infiltration of PMNs into corneal stroma, lipid peroxidation and keratocyte apoptosis. Environmental UV-B exposure should not be avoided after PRK. Amniotic membrane graft is beneficial to reduce keratocyte apoptosis and related corneal haze.

Effect of C1 esterase inhibitor on the cardiac dysfunction following ischemia and reperfusion in the isolated perfused rat heart

Complement-mediated neutrophil activation has been hypothesized to be an important mechanism of reperfusion injury. It has been proposed that C1 esterase inhibitor (C1 INH) may prevent the complement-dependent activation of polymorphonuclear leukocytes (PMNs) that occurs within postischemic myocardium. Therefore, The effect of C1 INH was examined in neutrophil dependent isolated perfused rat heart model of ischemia (I) (20 min) and reperfusion (R) (45 min). Administration of C1 INH (5 mg/Kg) to I/R hearts in the presence of PMNs (100 X 106) and homologous plasma improved coronary flow and preserved cardiac contractile function (p<0.001) in comparison to those I/R hearts receiving only vehicle. In addition, C1 INH significantly (p<0.001) reduced PMN accumulation in the ischemic myocardium as evidenced by an attenuation in myeloperoxidase activity. These findings demonstrate the C1 INH is a potent and effective cardioprotective agent inhibits leukocyte-endothelial interaction and preserves cardiac contractile function and coronary perfusion following myocardial ischemia and reperfusion.

Effect of Tetracycline-HCl on superoxide anion generation by polymorphonuclear leukocytes / 대한치주과학회지

It has been believed that the increased release of free oxygen radicals(O2-, H2O2 and OH-) might be one of important factors in the pathogenesis of periodontal diseases. Polymorphonuclear leukocytes(PMNs) constitute the primary host resistance factor against infection. They are prominent cells in both the gingival tissue and gingival sulcus in most forms of periodontal disease. In response to invading microorganisms, the activated PMNs and macrophages generate free oxygen radicals, which play an important role in bacterial killing. The normal production of these free oxygen radicals is vital for the successful resistance of individuals to bacterial infection. However, the enhanced production of reactive oxygen species by accumulating PMNs may be detrimental to the host in certain disease states. This study was performed to evaluate the effect of Tetracycline-HCl(Tc-HCl) on generation of superoxide anion by PMNs. For the present study, 60ml of peripheral venous blood were obtained from systemically healthy subjects by venipuncture in median cubital vein and PMNs were separated by a one-step Ficoll-Hypaque gradient centrifugation method. PMNs were incubated with 1microgram/ml P.gingivalis strain A7436 LPS, 5% serum and Tc-HCl(5, 10, 50 and 100microgram/ml). The superoxide anion analysis was carried out by superoxide dismutase-inhibitable cytochrome C reduction method using Microplate autoreader(BIO-TEK(TM) Instrument Inc.). The difference of superoxide anion generation between control group and Tc-HCl group was statistically analyzed by paired t-test. The superoxide anion generation in the course of time after treatment with Tc-HCl was analyzed by ANOVA, and the superoxide anion generation in the course of time after treatment with various concentrations of Tc-HCl was analyzed by Repeated Measurement test. The results were as follows: 1. Superoxide anion generation by PMNs was significantly decreased by Tc-HCl(p0.05). 3. Superoxide anion generation by PMNs was significantly decreased in the course of incubation time after treatment with Tc-HCl(p<0.05). The results demonstrate that the Tc-HCl inhibit superoxide anion generation by PMNs and the inhibitory effects depend on the exposure time rather than the concentration of Tc-HCl.

Time Course Change of Phagocytes and Proinflammatory activities in BALF in Endotoxin-induced Acute Lung Injury / 결핵

BACKGROUND: Severe acute lung injury(ALI), also known as the adult respiratory distress syndrome(ARDS), is a heterogenous nature of dynamic and explosive clinical synrome that exacts a mortality of approximately 50%. Endotoxin(ETX) is an abundant component of the outer membrane of gram-negative bacteria capable of inducing severe lung injury in gram-negative sepsis and gram-negative bacterial pneumonia, which are among the most common predisposing causes of ARDS. The influx of PMNs into airway tissue is a pathological hallmark of LPS-induced lung injury. And th3re is a substantial evidence suggesting that cytokines are important mediators of lung injury in gram-negative sepsis. However, the kinetics of phagocytes and cytokines by an exact time sequence and their respective pathogenic importance remain to be elucidated. This study was performed to investigate the role of phagocytes and proinflammatory cytokines in ETX-induced ALl through a time course of changes in the concentration of protein, TNFa and IL-6, and counts of total and its differential cells in BALF. The consecutive histologic findings were also evaluated. METHOD: The experimental animals, healthy male Sprague-Dawley, weighted 200+/-50g, were divided into controland ALI-group. ALI was induced by an intravenous administration of ETX, 5mg/kg. Above mentioned all parameters were examined at 0(control), 3, 6, 24, 72 h after administration of ETX. TNFa and IL-6 conc. in BALE were measured by a bioassay. RESULTS: The protein concentration and total leukocyte count(TC) in BALF was significantly increased at 3h compared to controls(p<0.05). The protein conc. was significantly elavated during observation period, but TC was significantly decreased at 72h(p<0.05 vs. 24h). There was a close relationship between TC and protein cone. in BALF(r = 0.65, p <0.001). The PMN and monocyte count was well correlated with TC in BALF, and the correlation of PMN(r=0.97, p<0.001) appeared to be more meaningful than that of monoeyte(r = 0.61, p<0.001). There was also a significant correlation between protein cone. and PMN or monocyte count in BALF(PMN vs. monocyte r = 0.55, p<0.005 vs. r = 0.64, p<0.001). The count of monocyte was significantly elavated during observation period though a meaningful reduction of PMN count in BALF at 72h, this observation suggested that monocyte may, at least, partipate in the process of lung injury steadly. In this sudy, there was no relationship between IL-6 and TNFt conc., and TNFa but not IL-6 was correlated with TC(r 0.61, p <0.05) and monocyte(r = 0.67, p<0.05) in BALF only at 3, 6h after ETX introduced. In particular, the IL-6 cone. increased earlier and rapidly peaked than TNFz cone. in BALF. In histologic findings, the cell counts of lung slices were increased from 3 to 72h(p<0.001 vs. NC). Alveolar wallthickness was increased from 6 to 24h(p<0.001 vs. NC). There was a significant correlation between the cell counts of lung slices and alveolar wall-thickness(r= 0.61, p<0.001). This result suggested that the cellular infiltrations might be followed by the alterations of interstitium, and the edematous change of alveolar wall might be most rapidly recovered to its normal condition in the process of repair. CONCLUSION: We concluded that although the role of PMIN is partly certain in ETX-induced ALI, it is somewhat inadequate to its known major impact on ALL Alveolar macrophage and/or non-immune cells such as pulmonary endothelial or epithelial cells, may be more importantly contributed to the initiation and perpetual progression of ETX-induced ALI. The IL-6 in ETX-induced ALI was independent to TNFa, measured by a bioassay in BALF. The early rise in IL-6 in BALF implies multiple origins of the IL-6.

Possible roles of LFA-1 and Fc gamma receptors on the functional immaturities of cord blood polymorphonuclear leukocytes

The functional immaturity of PMNs is one of the major causes of overwhelming sepsis in newborns. In this study, we observed functions and surface markers of PMNs to investigate what causes the functional immaturity of PMNs in newborns. As results, the percentage of EA rosette forming PMNs (58.5 +/- 15.5%) and the chemotactic movement (0.14 +/- 0.09 mm) of cord blood PMNs were significantly lower than those of adult peripheral blood PMNs (70.8 +/- 9.9%, 0.60 +/- 0.34 mm). Cord blood PMNs showed decreased glass adherence and ADCC activity. The expression of Fc gamma RII or Fc gamma RIII was a little lower than those of adult peripheral blood PMNs, but the expression of Fc gamma RI (43.1 +/- 26.8%) was significantly higher than that of adult peripheral blood PMNs (3.2 +/- 1.8%). There was a significant difference in LFA-1 expression between EA rosette forming PMNs (92.9 +/- 9.1%) and EA rosette non-forming PMNs (25.6 +/- 22.6%). From these results, it is assumed that neonatal PMNs may consist of heterogeneous populations. And the relatively high percentage of EA rosette non-forming PMNs which express a low level of LFA-1 may be responsible for the functional immaturity of cord blood PMNs.

The in Vivo Effects of Ethrane Anesthesia and Single Intravenous Injection of Hydrocortisone 250 mg on T-cell Subsets / 대한마취과학회지

The in vivo effects of ethrane and single intravenous injection of hydrocortisone 250 mg on T-cell subsets (T3, T4 and T8), s-cortisol, s-Na+ and K+, polymorphonuclear leukocytes, lymphocytes and HSR (T-helpers to T-suppressors ratio) has been studied. Six healthy volunteers and 16 patients was selected and observed. Sixteen patients has been done surgery under ethrane anesthesia, and single dose of hydrocortisone 250 mg was injected intravenously to eight patients among them. The result is as follows; 1) Serum cortisol level was increased after surgery under ethrane anesthesia and was decreased bellow control value at 24 hours after injection of hydrocortisone 250 mg. 2) After surgery under ethrane anesthesia, PMN's counts and monocyte counts in peripheral blood were increased, but absolute number of lymphocytes and T-cell subsets were decreased. 3) After surgery under ethrane anesthesia with hydrocortisone 250 mg, there was the most elevation of PMN's counts and the most redution of absolute number of T-cell subsets at 5 hours and was somewhat tended to return to control values, but remained changed at 24 hours, and monocyte counts was unchanged. 4) Adding hydrocortisone 250 mg compared with ethrane anesthesia alone, the increasing rate of PMN's counts and the decreasing rate of lymphocytes and absolute number of T-cell subsets were higher at 5 hours, and were lower at 24 hours. 5) Relative percentage of T-cell subsete and HSR were unchanged after surgery under ethrane anesthesia alone and adding hydrocortisone 250 mg.

Effects of polydatin Ⅳ on inhibiting respiratory burst of PMNs and scavenging oxygen free radicals / 中国药理学通报

Chemiluminescence method was used to measure: (1) active oxygen species generation induced by respiratory burst of polymor-phonuclear leucocytes (PMNs) from human blood stimulated with phorbol myristate acetate (PMA); (2) superoxide (O2) induced by xan-thine-xanthine oxidase system; (3) hydroxyl radicals ( ? OH ) generated by Vit C- Cu2+- zy-mosan; (4) the release of hydrogen peroxide (H2O2). Effects of polydatin IV on these active oxygen species were observed. The resultsshowed early stage of respiratory burst of PMNs was inhibited,but the later stage was delayed by polydatin IV, (2), (3) adn (4) were scavenged by polydatin IV and their median inhibitory concentrations (IC50?mol ? L-1) were 14.6,29.6 and 13.0 respectively. The results suggested that polydatin IV was a scavenger.