Processes In Spleen Research Articles

Regulating effects of triptriolide on the proliferation and apoptosis of splenic lymphocyte in vivo and in vitro

ObjectiveTriptriolide (T11) is isolated from the Chinese herb Tripterygium wilfordii Hook F (TwHF), which has been used as an immunosuppressive reagent for many years in China, while the role of this compound in TwHF is still unknown. In this study, we try to investigate the immune regulating effects of T11 on the proliferation and apoptosis of splenic lymphocyte in vivo and in vitro. Methodswe pretreated the BALB/c mice with T11 (2.8, 14, 28 mg/kg) and prednisone acetate (PA, 1.3 mg/kg, positive control) for 7 days before lipopolysaccharides (LPS, 3 mg/kg) stimulation. Then the histopathologic change of spleen was examined by H&E method, and the apoptosis of splenic lymphocyte was assessed by flow cytometry method. In addition, we further studied the inhibition effect of T11 (50, 100, 150, 200 µg/mL) on the proliferation and activity of splenic lymphocyte, which was with or without the stimulation of concanavalin A (Con A, 5 µg/mL) and lipopolysaccharides (LPS, 5 µg/mL) in vitro, respectively. ResultsWe found that T11 not only can restore the pathogenesis process in spleen, and boost the splenic lymphocyte apoptosis in vivo, but also can inhibit the splenic lymphocyte proliferation and affect its viability in vitro. ConclusionThe results indicated that T11 might modulate the immune function of LPS-induced BALB/c mice by inhibiting the proliferation and inducing the apoptosis of splenic lymphocyte. Therefore, T11 could be a novel immunomodulator in autoimmune disease treatment in clinical.

Role of SVEP1 in Stroma-Dependent Hematopoiesis In vitro.

Study of the microenvironment that supports hematopoietic stem cell (HSC) development in vivo is very difficult involving small numbers of interacting cells which are usually not well defined. While much is known about HSC niches located within the bone marrow in terms of contributing cell types and signalling molecules, very little is known about equivalent niches within spleen. Extramedullary hematopoiesis in spleen contributes myeloid cells important in the mobilisation of an immune response. As a result, it is important to develop in vitro models to identify the cells which constitute HSC niches in spleen and to identify the regulatory molecules supporting myeloid cell development. Studies described here document a model system to study the maintenance and differentiation of HSC by splenic stromal cells in vitro. The splenic stromal lines 5G3 and 3B5 differ in hematopoietic support capacity. SVEP1 and IGF2 are molecules of interest specifically expressed by 5G3 stroma. Gene knockdown technology using shRNA plasmids has been used to reduce gene expression in 5G3 and to determine specific effects on myeloid cell development following co-culture with overlaid hematopoietic progenitors in vitro. Knockdown of Svep1 gave specific inhibition of a dendritic cell (DC) population described previously in spleen (L-DC). Knockdown of Igf2 resulted in loss of production of a minor subset of conventional (c) DC. SVEP1 is now considered a marker of mesenchymal stromal cells with osteogenic differentiative capacity reflective of perivascular stromal cells. The power of this in vitro model is evidenced by the fact that it has been used to define SVEP1 as a specific adhesion molecule that regulates the hematopoietic process dependent on stromal niche interaction. The identification of stromal cells and molecules that contribute to the hematopoietic process in spleen, brings us closer to the realm of therapeutically regulating hematopoiesis in vivo, and to inhibiting niches which support cancer stem cells.

Acute toxicity of C60–Cis-Pt nanocomplex in vivo

Acute toxicity of noncovalent nanocomplex of C60 fullerene with Cisplatin (Cis-Pt) (the ratio of compounds C60:Cis-Pt by weight 1:1) in a dose progression (7.5 + 7.5, 15.0 + 15.0, 22.5 + 22.5, 30.0 + 30.0, 37.5 + 37.5, and 45.0 + 45.0 mg/kg) was investigated in mice of BALB/c line. Mice were injected with C60–Cis-Pt nanocomplex or free Cis-Pt at the same doses single intraperitoneally and observed for 14 days. A comparative study of the toxicity of free Cis-Pt and Cis-Pt at noncovalent complexation with C60 fullerene (C60–Cis-Pt nanocomplex) demonstrated halving the toxicity of C60–Cis-Pt nanocomplex in comparison with free Cis-Pt: LD50 (Cis-Pt) 15.6 ± 1.3 mg/kg vs LD50 (C60–Cis-Pt) 36.1 ± 2.8 mg/kg. The toxic effects of C60–Cis-Pt nanocomplex and free Cis-Pt were detected at doses 22.5 + 22.5 mg/kg and 15.0 mg/kg, respectively, and were accompanied by impaired mice behavior, decreased body weight and animal survival, hematotoxicity, and pathological changes in heart, liver, spleen, and kidneys. It was shown that C60 fullerene complexed with Cis-Pt prevented drug-induced decrease in animal survival, anemia, thrombocytosis, and leukopenia development, as well as inflammatory processes in spleen, heart, kidney, and liver. The obtained results indicate the prospects for using noncovalent nanocomplex C60–Cis-Pt in antitumor therapy at low therapeutic doses of Cis-Pt.

Protective Effect of Casperome®, an Orally Bioavailable Frankincense Extract, on Lipopolysaccharide- Induced Systemic Inflammation in Mice.

Introduction: Despite recent advances in critical care, sepsis remains a crucial cause of morbidity and mortality in intensive care units. Therefore, the identification of new therapeutic strategies is of great importance. Since ancient times, frankincense is used in traditional medicine for the treatment of chronic inflammatory disorders such as rheumatoid arthritis. Thus, the present study intends to evaluate if Casperome® (Casp), an orally bioavailable soy lecithin-based formulation of standardized frankincense extract, is able to ameliorate systemic effects and organ damages induced by severe systemic inflammation using a murine model of sepsis, i.e., intraperitoneal administration of lipopolysaccharides (LPS).Methods: Male 60-day-old mice were assigned to six treatment groups: (1) control, (2) LPS, (3) soy lecithin (blank lecithin without frankincense extract), (4) Casp, (5) soy lecithin plus LPS, or (6) Casp plus LPS. Soy lecithin and Casp were given 3 h prior to LPS treatment; 24 h after LPS administration, animals were sacrificed and health status and serum cytokine levels were evaluated. Additionally, parameters representing liver damage or liver function and indicating oxidative stress in different organs were determined. Furthermore, markers for apoptosis and immune cell redistribution were assessed by immunohistochemistry in liver and spleen.Results: LPS treatment caused a decrease in body temperature, blood glucose levels, liver glycogen content, and biotransformation capacity along with an increase in serum cytokine levels and oxidative stress in various organs. Additionally, apoptotic processes were increased in spleen besides a pronounced immune cell infiltration in both liver and spleen. Pretreatment with Casp significantly improved health status, blood glucose values, and body temperature of the animals, while serum levels of pro-inflammatory cytokines and oxidative stress in all organs tested were significantly diminished. Finally, apoptotic processes in spleen, liver glycogen loss, and immune cell infiltration in liver and spleen were distinctly reduced. Casp also appears to induce various cytochromeP450 isoforms, thus causing re-establishment of liver biotransformation capacity in LPS-treated mice.Conclusion: Casp displayed anti-inflammatory, anti-oxidative, and hepatoprotective effects. Thus, orally bioavailable frankincense extracts may serve as a new supportive treatment option in acute systemic inflammation and accompanied liver dysfunction.

Redox balance in white rats’ spleen under induced carcinogenesis

Features of lipid peroxidation processes and antioxidant system in spleen tissue of white rats in experimental carcinogenesis and application of cytostatics were studied in experimental research. In DMH-induced cancerogenesis it was found a significant increase concentration of TBA-reactive substances and lipid hydroperoxides, decrease of antioxidant enzyme activity and reduced glutathione concentration in the examined organ tissue. The combined use of chemotherapeutic drugs increased imbalance of redox processes in spleen.

IMMUNOACTIVITY PROPERTIES OF ASCIDIAN EXTRACT ON ACUTE TOXIC HEPATITIS MODEL INVESTIGATION

Immunodepressive state risk at patient with toxic damage is one of the reasons for the search of the remedy with hepatoprotective and immunoregulation properties. There was established that extract of marine hydrobiont Halocynthia aurantium (AE) possess the hepato-and membranoprotection effects. Immunoactivity properties of AE on the background of the experimental acute toxic hepatitis caused by carbon tetrachloride (CCl4 50% in olive oil, 2 ml/kg during 4 days) at rats Wistar (40 animals, male) were investigated. It was known, that CCl4 action the metabolically and immune process disturbance in animals manifested. It was shown that 7 days after CCl4 action there did not any positive changes in the peripheral blood cells and lymphoid organs of animals. This data was shown that negative consequence of toxic influence at rats continued. АЕ administration (24 mg/kg) during 7days after CCl4 action provided positive effect on the leukocytes and lymphocytes population at rats. There was established that CD8+ higher level decrease (on 25-30%, p≤0,05), CD4+ quantity increase. Moreover there was stimulated T-helper lymphocytes activity and recovered the immunity reactions by optimization T-helper/T-suppressor lymphocytes correlation and this index approaches to the normal condition in blood and spleen under AE influence. The depressed by CCl4 action lymphoproliferative processes in spleen and thymus was restoration to intact level in animals with AE. And there was pronounced stimulating effect on the nonspecific immunoglobulins synthesis too. These results possibly are the evidence of B-lymphocyte functional activity increased and humoral reactivity intensify by AE. The positive effect of AE may be explained both direct action on cellular and humoral parameters of immunity and promote the restoration of conditions for normal interaction between different parts of immune system. Thus, this data was demonstrated the potential use of AE for immunity correction in acute toxic hepatitis conditions.

The Morphological Changes in the Internal Organs in Tumor-Bearing Rats at Intravenous Injection of Citrate-Stabilized Magnetite Nanoparticles

The usefulness of magnetite nanoparticles to improve contrast in magnetic resonance imaging (MRI) of tumor visualization was actively investigated currently. In this work, we conducted MRI and morphological examination of internal organs of 30 white outbred rats with transplanted liver tumor PC-1 after intravenous administration of magnetite hydrosols. Intravenous administration of citrate-stabilized magnetite nanoparticles in a dosage of 20 μg/kg and 16 mg/kg resulted in a disruption of blood filling of the organs, mainly due to the plethora. The dystrophic cell damage was observed in liver and in kidneys. The activation of immune processes in spleen was fixed after intravenous administration of citrate stabilized magnetite nanoparticles. In both doses, the nanoparticles were founded in the heart and brain, but the significant iron accumulation was detected in the tumor by methods of MRI and atomic absorption spectroscopy (AAS) only in dosage of 16 mg/kg. It is possible to conclude that selected dosage is sufficient for accumulation of nanoparticles in tumor, but it does not cause significant changes in the internal organs.

Genomic sequences comparison and differential expression of miiuy croaker MHC class I gene, after infection by Vibrio anguillarum

Major histocompatibility complex (MHC) has a central role in the adaptive immune system by presenting foreign peptide to the T-cell receptor. MHC gene family contains two main subgroups of immunologically active molecules. In order to study the molecular function and genomic characteristic of class I gene in teleost, the full lengths of MHC class Iα cDNA and genomic sequence were cloned from miiuy croaker ( Miichthys miiuy). Seven exons and six introns were identified in miiuy croaker class Iα gene. This genomic structural feature of miiuy croaker is similar to that present in some fishes such as Japanese flounder and Atlantic salmon, but different from that present in some other fishes such as half-smooth tongue sole and channel catfish. The deduced amino acid sequence of class Iα gene had 25.9–54.1% identity with those of mammal and teleost. Real-time quantitative RT-PCR demonstrated that the MHC class Iα gene was ubiquitously expressed in 10 normal tissues; expression levels of MHC Iα gene were found first upregulated and then downregulated throughout the pathogenic bacteria infection process in spleen and kidney.

α1- and β-Adrenoblockers Effects on Immunogenesis in Rats under Thermoneutral Conditions and after Cooling of Various Extent

Experiments with rats showed that ionophoretic delivery of alpha1-adrenoblocker into the skin does not change effects of superficial and deep cooling and did not affect the amount of the antibody producing cells in spleen and a blood level of circulating antibodies under thermoneutral conditions. Meanwhile alpha1-adrenoblocker abolished inhibitory effect of fast deep cooling on antigen-binding properties of spleen cells and peritoneal extraction cells, and markedly stimulated it. beta-Adrenoblocker had no effect on immunogenesis under thermoneutral conditions and changes modulating effect of cooling on immune response. It abolished immunosuppressive effect of deep cooling and enhanced the stimulating effect of superficial cooling. It was true both for antigen-binding processes in spleen and peritoneal cavity and for antibody production in spleen. The results obtained indicate an involvement of alpha1--and beta-adrenoreceptors in immune response inhibition after deep cooling. Adrenoblockers effects on various immune competent cells are ambiguous and depend on temperature.

Beneficial pleiotropic actions of melatonin in an experimental model of septic shock in mice: regulation of pro‐/anti‐inflammatory cytokine network, protection against oxidative damage and anti‐apoptotic effects

Septic shock, the most severe problem of sepsis, is a lethal condition caused by the interaction of a pathogen-induced long chain of sequential intracellular events in immune cells, epithelium, endothelium, and the neuroendocrine system. The lethal effects of septic shock are associated with the production and release of numerous pro-inflammatory biochemical mediators including cytokines, nitric oxide and toxic oxygen and nitrogen radicals, together with development of massive apoptosis. As melatonin has remarkable properties as a cytokine modulator, antioxidant and anti-apoptotic agent, the present study was designed to evaluate the possible protective effect of melatonin against LPS-induced septic shock in Swiss mice. We observed that intraperitoneally (i.p.) administered-melatonin (10 mg/kg) 30 min prior, and 1 hr after i.p. LPS injection (0.75 mg/animal) markedly protected mice from the LPS lethal effects with 90% survival rates for melatonin and 20% for LPS-injected mice after 72 hr. The melatonin effect was mediated by modulating the release of pro-/anti-inflammatory cytokine levels, protection from oxidative damage and counteracting apoptotic cell death. Melatonin was able to partially counteract the increase in LPS-induced pro-inflammatory cytokine levels such as tumor necrosis factor-alpha, IL-12 and interferon-gamma at the local site of injection, while it increased the production of the anti-inflammatory cytokine IL-10 both locally and systemically. Furthermore, melatonin inhibited the LPS-induced nitrite/nitrate and lipid peroxidation levels in brain and liver and counteracted the sepsis-associated apoptotic process in spleen. In conclusion, we have demonstrated that melatonin improves the survival of mice with septic shock via its pleiotropic functions as an immunomodulator, antioxidant and anti-apoptotic mediator.